Korean Amberjack Skin-Inspired Hyaluronic Acid Bioink for Reconstruction of Human SkinClick to copy article linkArticle link copied!
- Hoai-Thuong Duc BuiHoai-Thuong Duc BuiDepartment of Medical Biomaterials Engineering, Kangwon National University, Chuncheon 24341, Republic of KoreaMore by Hoai-Thuong Duc Bui
- Wanho ChoWanho ChoDepartment of Medical Biomaterials Engineering, Kangwon National University, Chuncheon 24341, Republic of KoreaMore by Wanho Cho
- Jae Keun ParkJae Keun ParkDepartment of Medical Biomaterials Engineering, Kangwon National University, Chuncheon 24341, Republic of KoreaMore by Jae Keun Park
- Moon Sue Lee
- Hong Kee Kim
- Hyuk Sang Yoo*Hyuk Sang Yoo*Email: [email protected]Department of Medical Biomaterials Engineering, Kangwon National University, Chuncheon 24341, Republic of KoreaKangwon Radiation Convergence Research Support Center, Kangwon National University, Chuncheon 24341, Republic of Korealnstitute of Bioscience & Biotechnology, Kangwon National University, Chuncheon 24341, Republic of Korealnstitute of Molecular Science and Fusion Technology, Kangwon National University, Chuncheon 24341, Republic of KoreaMore by Hyuk Sang Yoo
Abstract
Decellularized extracellular matrix (dECM) has been extensively employed as tissue engineering scaffolds because its components can greatly enhance the migration and proliferation of cultivating cells. In this study, we decellularized Korean amberjack skin and incorporated soluble fractions in hyaluronic acid hydrogels with 3D-printed tissue engineering hydrogels to overcome any limitation of animal-derived dECM. The hydrolyzed fish-dECM was mixed with methacrylated hyaluronic acid and chemically crosslinked to 3D-printed fish-dECM hydrogels, where fish-dECM contents affected both printability and injectability of the hydrogels. Swelling ratios and mass erosion of the 3D-printed hydrogels were dependent on fish-dECM contents, where higher fish-dECM in the hydrogel increased swelling ratios and mass erosion rates. The higher content of fish-dECM considerably enhanced the viability of the incorporated cells in the matrix for 7 days. Artificial human skin was constructed by seeding human dermal fibroblasts and keratinocytes in the 3D-printed hydrogels, and a formation of a bilayered skin was visualized with tissue staining. Thus, we envision that 3D-printed hydrogels containing fish-dECM can be an alternative bioink composed of a non-mammal-derived matrix.
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1. Introduction
2. Results and Discussion
Figure 1
Figure 1. Schematic diagram of the preparation of solubilized fish-dECM and 3D hybrid bioink for cocultivation of HDF and HaCaT cells with a 3D-printed model in an air–liquid condition. (a) Solubilization process of fish-dECM in alkaline conditions. (b) Preparation of 3D hybrid bioink (pre-gel) and fabrication of a 3D-printed model via 3D printing and secondary crosslinking of the 3D scaffold.
total protein (μg/mL) | |||||
---|---|---|---|---|---|
fish-dECM solubilization time (hours) | initial weight of fish-dECM (mg) | unsolubilized fish-dECM (mg) | solubilized fish-dECM (mg)a | before dialysis | after dialysis |
4 | 1000 | 30.7 ± 5.6 | 211.4 ± 6.8 | 741.1 ± 34.0 | 116.2 ± 1.4 |
12 | 32.0 ± 2.5 | 291.7 ± 9.6 | 434.7 ± 87.9 | 157.7 ± 2.5 | |
24 | 12.2 ± 1.4 | 182.3 ± 6.5 | 305.6 ± 43.2 | 172.0 ± 3.7 | |
48 | 11.1 ± 1.3 | 73.8 ± 14.7 | 246.7 ± 51.8 | 136.8 ± 3.6 |
Weight of solubilized fish-dECM after dialysis.
Figure 2
Figure 2. Characterization of dECM. (a) UV–vis spectra of solubilized fish-dECM before or after dialysis. Absorbance was normalized to total protein determined by a BCA assay. (b) Cytotoxicity of fish-dECM eluant. Fish-dECM (6 cm2) was eluated with DMEM supplemented with 10% of FBS (1 mL), followed by mixing with cell cultivation medium at different mixing ratios (0–100%) and addition to NIH3T3 cells (1 × 104 cells/well (surface area = 0.32 cm2)). After 24 h, an MTT-based cytotoxicity assay was performed and the viability of the treated cells was normalized with respect to those without treatment.
Figure 3
Figure 3. Investigation of injectability and printability of different HA-based bioinks, and characterization of 3D-printed models. (a, b) Optimization of 3D hybrid bioink. Injectability and printability of 3D hybrid bioink crosslinked at various DVS molar ratios (molar ratio of HA:DVS = 1:0.020–0.055) were examined (O, high; Δ, medium; X, low injectability/printability). (c) Images of the custom design model and 3D-printed model (scale bar = 1 cm). (d) Water swelling ratio of the 3D-printed model with 7% (v/v) DVS (n = 3). (e) Mass erosion of 3D-printed model; 3D-printed model was immersed in PBS (pH 7.4) at 37 °C for 7, 14, and 21 d. At the designated time point, the degraded 3D model was washed thrice with DW and freeze-dried; the dried 3D model weight was recorded (n = 3). *indicates a significant difference between HA/0dECM, HA/10dECM, and HA/30dECM; p < 0.05 were considered statistically significant.
Figure 4
Figure 4. In vitro studies on cell attachment and cell viability of HA-based 3D-printed scaffolds. (a) NIH3T3 cells attachment on 3D-printed models. Unbound cells were stained with trypan blue and counted using a hemocytometer. Cell attachment* (%) = ((number of initial cells – counted cells)/number of initial cells × 100). (b) Proliferation of cells cultivated on 3D-printed models for 3, 5, and 7 d (n = 3). Cell proliferation was determined by a WST-1-based colorimetric assay. * indicates a significant difference between HA/0dECM, HA/10dECM, and HA/30dECM; p < 0.05 were considered statistically significant.
Figure 5
Figure 5. HDF and HaCaT cocultivation on 3D-printed scaffolds. (a) Cocultivation of the HDF/HaCaT/3D-printed model. After 5 d of incubation with HDF cells, HaCaT cells were added and incubated for another 5 days in air–liquid conditions to induce keratinization of HaCaT cells. (b) Masson’s trichrome and hematoxylin staining of HDF and HaCaT-cocultivated 3D scaffolds. Collagens (white arrows) and cell nuclei (white circles) are stained in blue and violet. Dashed lines separate the regions of the epidermis (ED) and the dermis (D) (scale bar = 100 μm).
3. Conclusions
4. Experimental Section
4.1. Materials
4.2. Preparation of Fish-dECM
4.3. Preparation of Solubilized Fish-dECM
4.4. Preparation of HA-Based Hybrid Ink
4.5. 3D Printing
4.6. Characterization of the 3D-Printed Model
4.7. In Vitro Cell Cultivation
4.8. In Vitro Construction of Bilayered Human Skin
4.9. Statistical Analysis
Supporting Information
The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.3c01642.
UV–vis spectra of solubilized fish-dECM before or after dialysis (Figure S1) and the water swelling ratio of 3D-printed models with 5 and 10% DVS (Figure S2) (PDF)
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References
This article references 54 other publications.
- 1Kiyotake, E. A.; Douglas, A. W.; Thomas, E. E.; Nimmo, S. L.; Detamore, M. S. Development and Quantitative Characterization of the Precursor Rheology of Hyaluronic Acid Hydrogels for Bioprinting. Acta Biomater. 2019, 95, 176– 187, DOI: 10.1016/j.actbio.2019.01.041Google Scholar1https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvFeksrY%253D&md5=fdae8ca8387f8c70ad58c7ddfd713dcbDevelopment and quantitative characterization of the precursor rheology of hyaluronic acid hydrogels for bioprintingKiyotake, Emi A.; Douglas, Alexander W.; Thomas, Emily E.; Detamore, Michael S.Acta Biomaterialia (2019), 95 (), 176-187CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Bioprinting technologies have tremendous potential for advancing regenerative medicine due to the precise spatial control over depositing a printable biomaterial, or bioink. Despite the growing interest in bioprinting, the field is challenged with developing biomaterials for extrusion-based bioprinting. The paradigm of contemporary bioink studies relies on trial-and-error methods for discovering printable biomaterials, which has little practical use for others who endeavor to develop bioinks. There is pressing need to follow the precedent set by a few pioneering studies that have attempted to standardize bioink characterizations for detg. the properties that define printability. Here, we developed a pentenoate-functionalized hyaluronic acid hydrogel (PHA) into a printable bioink and used three recommended, quant. rheol. assessments to characterize the printability: (1) yield stress, (2) viscosity, and (3) storage modulus recovery. The most important characteristic is the yield stress; we found a yield stress upper limit of ∼1000 Pa for PHA. Measuring the viscosity was advantageous for detg. shear-thinning behavior, which aided in extruding highly viscous PHA through a nozzle. Post-printing recovery is required to maintain shape fidelity and we found storage modulus recoveries above ∼85% were sufficient for PHA. Two formulations had superior printability (i.e., 1.5 MDa PHA - 4 wt%, and 1 MDa PHA - 8 wt%), and increasing cell concns. in PHA up to 9 × 106 cells/mL had minimal effects on the printability. Even so, other factors such as sterilization and peptide modifications to enhance bioactivity may influence printability, highlighting the need for investigators to consider such factors when developing new bioinks. Bioprinting has potential for regenerating damaged tissues; however, there are a limited no. of printable biomaterials, and developing new bioinks is challenging because the required material phys. properties for extrusion-based printing are not yet known. Most new bioinks are developed by trial-and-error, which is neither efficient nor comparable across materials. There is a need for the field to begin utilizing std. methods proposed by a few pioneering studies to characterize new bioinks. Therefore, we have developed the printability of a hyaluronic acid based-hydrogel and characterized the material with three quant. rheol. tests. The current work impacts the bioprinting field by demonstrating and encouraging the use of universal bioink characterizations and by providing printability windows to advance new bioink development.
- 2Ge, B.; Wang, H.; Li, J.; Liu, H.; Yin, Y.; Zhang, N.; Qin, S. Comprehensive Assessment of Nile Tilapia Skin. Mar. Drugs 2021, 18, 178– 194, DOI: 10.3390/md18040178Google ScholarThere is no corresponding record for this reference.
- 3Ouyang, L.; Highley, C. B.; Rodell, C. B.; Sun, W.; Burdick, J. A. 3D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-Linking. ACS Biomater. Sci. Eng. 2016, 2, 1743– 1751, DOI: 10.1021/acsbiomaterials.6b00158Google Scholar3https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xos1Skt7c%253D&md5=870810033017a9e057780d8e4f861e963D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-LinkingOuyang, Liliang; Highley, Christopher B.; Rodell, Christopher B.; Sun, Wei; Burdick, Jason A.ACS Biomaterials Science & Engineering (2016), 2 (10), 1743-1751CODEN: ABSEBA; ISSN:2373-9878. (American Chemical Society)The development of printable biomaterial inks is crit. to the application of 3D printing in biomedicine. To print high-resoln. structures with fidelity to a computer-aided design, materials used in 3D printing must be capable of being deposited on a surface and maintaining a printed structure. A dual-crosslinking hyaluronic acid system was studied here as a printable hydrogel ink, which encompassed both shear-thinning and self-healing behaviors via guest-host bonding, as well as covalent crosslinking for stabilization using photopolymn. When either guest-host assembly or covalent crosslinking was used alone, long-term stable structures were not formed, because of network relaxation after printing or dispersion of the ink filaments prior to stabilization, resp. The dual-crosslinking hydrogel filaments formed structures with greater than 16 layers that were stable over a month with no loss in mech. properties and the printed filament size ranged from 100 to 500 μm, depending on printing parameters (needle size, speed, and extrusion flux). Printed structures were further functionalized (i.e., RGD peptide) to support cell adhesion. This work highlights the importance of ink formulation and crosslinking on the printing of stable hydrogel structures.
- 4Petta, D.; Armiento, A. R.; Grijpma, D.; Alini, M.; Eglin, D.; D’Este, M. 3D Bioprinting of a Hyaluronan Bioink through Enzymatic-and Visible Light-Crosslinking. Biofabrication 2018, 10, 044104 DOI: 10.1088/1758-5090/aadf58Google Scholar4https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXmtFSitr0%253D&md5=c99806172bc4676bb93f6b30d6b3ed6cThree dimensional bioprinting of a hyaluronan bioink through enzymatic-and visible light-crosslinkingPetta, D.; Armiento, A. R.; Grijpma, D.; Alini, M.; Eglin, D.; D'Este, M.Biofabrication (2018), 10 (4), 044104/1-044104/11CODEN: BIOFFN; ISSN:1758-5090. (IOP Publishing Ltd.)Extrusion-based three-dimensional bioprinting relies on bioinks engineered to combine viscoelastic properties for extrusion and shape retention, and biol. properties for cytocompatibility and tissue regeneration. To satisfy these conflicting requirements, bioinks often utilize either complex mixts. or complex modifications of biopolymers. In this paper we introduce and characterize a bioink exploiting a dual crosslinking mechanism, where an enzymic reaction forms a soft gel suitable for cell encapsulation and extrusion, while a visible light photo-crosslinking allows shape retention of the printed construct. The influence of cell d. and cell type on the rheol. and printability properties was assessed correlating the printing outcomes with the damping factor, a rheol. characteristic independent of the printing system. Stem cells, chondrocytes and fibroblasts were encapsulated, and their viability was assessed up to 14 days with live/dead, alamar blue and trypan blue assays. Addnl., the impact of the printing parameters on cell viability was investigated. Owing to its straightforward prepn., low modification, presence of two independent crosslinking mechanisms for tuning shear-thinning independently of the final shape fixation, the use of visible green instead of UV light, the possibility of encapsulating and sustaining the viability of different cell types, the hyaluronan bioink here presented is a valid biofabrication tool for producing 3D printed tissue-engineered constructs.
- 5Petta, D.; D’Amora, U.; Ambrosio, L.; Grijpma, D. W.; Eglin, D.; D’Este, M. Hyaluronic Acid as a Bioink for Extrusion-Based 3D Printing. Biofabrication 2020, 12, 032001 DOI: 10.1088/1758-5090/ab8752Google Scholar5https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXitFCisr%252FN&md5=afa69e4f9418987ed3741ae1665a74aeHyaluronic acid as a bioink for extrusion-based 3D printingPetta, D.; D'amora, U.; Ambrosio, L.; Grijpma, D. W.; Eglin, D.; D'este, M.Biofabrication (2020), 12 (3), 032001CODEN: BIOFFN; ISSN:1758-5090. (IOP Publishing Ltd.)A review. Biofabrication is enriching the tissue engineering field with new ways of producing structurally organized complex tissues. Among the numerous bioinks under investigation, hyaluronic acid (HA) and its derivs. stand out for their biol. relevance, cytocompatibility, shear-thinning properties, and potential to fine-tune the desired properties with chem. modification. In this paper, we review the recent advances on bioinks contg. HA. The available literature is presented based on subjects including the rheol. properties in connection with printability, the chem. strategies for endowing HA with the desired properties, the clin. application, the most advanced preclin. studies, the advantages and limitations in comparison with similar biopolymer-based bioinks, and future perspectives.
- 6Qiao, Y.; Xu, S.; Zhu, T.; Tang, N.; Bai, X.; Zheng, C. Preparation of Printable Double-Network Hydrogels with Rapid Self-Healing and High Elasticity Based on Hyaluronic Acid for Controlled Drug Release. Polymer 2020, 186, 121994 DOI: 10.1016/j.polymer.2019.121994Google Scholar6https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXit1CmurzL&md5=183ff4e2d9197f1926fd150df38e3b1aPreparation of printable double-network hydrogels with rapid self-healing and high elasticity based on hyaluronic acid for controlled drug releaseQiao, Yang; Xu, Shichao; Zhu, Tianzhe; Tang, Nan; Bai, Xuejian; Zheng, ChunmingPolymer (2020), 186 (), 121994CODEN: POLMAG; ISSN:0032-3861. (Elsevier Ltd.)On account of the features about computer-accurate control, the technol. of 3D printing has a broad future for developing customized ergonomic biomedical materials. Herein, a printable dual-network hydrogel for drug delivery was designed. The acrylamide-modified hyaluronic acid was mixed with folic acid and Fe3+ at first. With the formation of metal-carboxylate coordination bonds, the first layer of phys. crosslinking network (HFF) was formed, followed by performing the spectral anal. and rheol. measurements to verify the bonding. The second layer of PAM network was then polymd. by UV radiation, with high elasticity and fatigue resistance to satisfy its application as a wound dressing. Furthermore, acetylsalicylic acid (Asa) was used as a drug model resulted in a pH responsiveness of the prepd. hydrogels with the sustained drug releasing behavior over 300 h. These results signified the potential application of this self-healing hydrogel with good mech. properties in regenerative medicine.
- 7Antich, C.; de Vicente, J.; Jiménez, G.; Chocarro, C.; Carrillo, E.; Montañez, E.; Gálvez-Martín, P.; Marchal, J. A. Bio-Inspired Hydrogel Composed of Hyaluronic Acid and Alginate as a Potential Bioink for 3D Bioprinting of Articular Cartilage Engineering Constructs. Acta Biomater. 2020, 106, 114– 123, DOI: 10.1016/j.actbio.2020.01.046Google Scholar7https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjtVanu74%253D&md5=8df42b62707737a82d3a9687849b11c4Bio-inspired hydrogel composed of hyaluronic acid and alginate as a potential bioink for 3D bioprinting of articular cartilage engineering constructsAntich, Cristina; de Vicente, Juan; Jimenez, Gema; Chocarro, Carlos; Carrillo, Esmeralda; Montanez, Elvira; Galvez-Martin, Patricia; Marchal, Juan AntonioActa Biomaterialia (2020), 106 (), 114-123CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Bioprinting is a promising tool to fabricate well-organized cell-laden constructs for repair and regeneration of articular cartilage. In this study, we focused on the use of one of the main cartilage components, hyaluronic acid (HA), to design and formulate a new bioink for cartilage tissue 3D bioprinting. To produce cartilage constructs with optimal mech. properties, HA-based bioink was co-printed with polylactic acid (PLA). HA-based bioink was found to improve cell functionality by an increase in the expression of chondrogenic gene markers and specific matrix deposition and, therefore, tissue formation. It is a promising bioink candidate for cartilage tissue engineering based in 3D bioprinting. The recent appearance of 3D printing technol. has enabled great advances in the treatment of osteochondral disorders by fabrication of cartilage tissue constructs that restore and/or regenerate damaged tissue. In this study, we describe for first time the development of a bioink based on the main component of cartilage, HA, with suitable biol. and mech. properties, without involving toxic procedure, and its application in cartilage tissue bioprinting. Hybrid constructs prepd. by co-printing this bioink and thermoplastic polymer PLA provided an optimal niche for chondrocyte growth and maintenance as well as mech. properties necessary to support load forces exerted in native tissue. We highlight the translation potential of this HA-based bioink in the clin. arena.
- 8Roushangar Zineh, B.; Shabgard, M. R.; Roshangar, L.; Jahani, K. Experimental and Numerical Study on the Performance of Printed Alginate/Hyaluronic Acid/Halloysite Nanotube/Polyvinylidene Fluoride Bio-Scaffolds. J. Biomech. 2020, 104, 109764 DOI: 10.1016/j.jbiomech.2020.109764Google Scholar8https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB38zgvFGksQ%253D%253D&md5=b5e5f522b57a0b2a4029939280f869b7Experimental and numerical study on the performance of printed alginate/hyaluronic acid/halloysite nanotube/polyvinylidene fluoride bio-scaffoldsRoushangar Zineh Babak; Jahani Kamal; Shabgard Mohammad Reza; Roshangar LeilaJournal of biomechanics (2020), 104 (), 109764 ISSN:.The growing usage of printed bio scaffolds in the field of regenerative medicine has made this field very important in biomedical engineering. In this regard, three-dimensional printing (3D) technique needs bio-materials with higher mechanical and biological performance. The biomaterials with high mechanical performance beside its bio compatibility are limited. A novel bio-material made of Alginate, Hyaluronic acid, Halloysite Nanotube and Polyvinylidene Fluoride was used and characterized for printing cartilage bio scaffolds through numerical studies. CaCl2 was used for crosslinking of biomaterial. Scanning Electron Microscopy, mechanical tests (tensile and compressive test), MTT assay were conducted for evaluating this novel biomaterial. Different structures of bio material were simulated for numerical studies. The numerical study was performed in ANSYS 18 using three parameter Mooney-Rivlin model. According to experimental and numerical results, Halloysite Nanotube increases the tensile and compressive strength of biomaterial up to 47%. Results show that biomaterial have good mechanical performance due to mechanical forces required for cartilage bio scaffolds besides its high biological performance. Polyvinylidene fluoride reduces the mechanical performance while increasing the cell viability. MTT assay results performed on day 0, day 2 and day 6 show increase in cell number to be about twice for biomaterial containing 40 mg/ml alginate, 40 mg/ml halloysite nanotube, 10 mg/ml hyaluronic acid and 1 w/v Polyvinylidene fluoride. Numerical simulation shows high mechanical performance of bio material in different scaffolds structure. The best structure of bio scaffolds was achieved with 0.4 mm nozzle diameter and 0.4 space between rows.
- 9Pérez, L. A.; Hernández, R.; Alonso, J. M.; Pérez-González, R.; Sáez-Martínez, V. Hyaluronic Acid Hydrogels Crosslinked in Physiological Conditions: Synthesis and Biomedical Applications. Biomedicines 2021, 9, 1113 DOI: 10.3390/biomedicines9091113Google Scholar9https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXisVyqtb3I&md5=8bc04f98b8ce3998dccf9808ca1149f7Hyaluronic Acid Hydrogels Crosslinked in Physiological Conditions: Synthesis and Biomedical ApplicationsPerez, Luis Andres; Hernandez, Rebeca; Alonso, Jose Maria; Perez-Gonzalez, Raul; Saez-Martinez, VirginiaBiomedicines (2021), 9 (9), 1113CODEN: BIOMID; ISSN:2227-9059. (MDPI AG)Hyaluronic acid (HA) hydrogels display a wide variety of biomedical applications ranging from tissue engineering to drug vehiculization and controlled release. To date, most of the com. available hyaluronic acid hydrogel formulations are produced under conditions that are not compatible with physiol. ones. This review compiles the currently used approaches for the development of hyaluronic acid hydrogels under physiol./mild conditions. These methods include dynamic covalent processes such as boronic ester and Schiff-base formation and click chem. mediated reactions such as thiol chem. processes, azide-alkyne, or Diels Alder cycloaddn. Thermoreversible gelation of HA hydrogels at physiol. temp. is also discussed. Finally, the most outstanding biomedical applications are indicated for each of the HA hydrogel generation approaches.
- 10Yang, R.; Tan, L.; Cen, L.; Zhang, Z. An Injectable Scaffold Based on Crosslinked Hyaluronic Acid Gel for Tissue Regeneration. RSC Adv. 2016, 6, 16838– 16850, DOI: 10.1039/c5ra27870hGoogle Scholar10https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xit1Orsb4%253D&md5=c915a516700130b50ceeae0721fa2660An injectable scaffold based on crosslinked hyaluronic acid gel for tissue regenerationYang, Rui; Tan, Linhua; Cen, Lian; Zhang, ZhibingRSC Advances (2016), 6 (20), 16838-16850CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)Injectable scaffolds have great potential in specialised applications in regenerative medicine. In this study, hyaluronic acid hydrogels (HAGs) were prepd. by crosslinking hyaluronic acid (HA) with 1,4-butanediol diglycidyl ether (BDDE). Applications of HAG as an injectable scaffold for regenerating functional tissues were proposed by matching its viscoelastic properties with those of biol. tissues. The effect of BDDE concn. on different properties of HAGs was explored. Swelling properties, cross-sectional morphol., and BDDE residues of the resulting gels were investigated. Rheol. properties of different HAGs were measured by monitoring their storage modulus (G') and loss modulus (G'') and compared with those of biol. tissues. It was shown that HAGs (BDDE from 0.4 vol% to 1.0 vol%) possess great water absorbing capability with swelling ratios ranging from 99.7 to 78.9. The higher the concn. of the crosslinker used, the more rigid the resulting hydrogel, subsequently the lower the swelling ratio would be and the higher the G' and G'' values as well. Similar viscoelastic behaviors were found between HAGs and biol. tissues, such as epidermis, dermis, articular cartilage and tooth germ. SEM revealed that HAG obtained at 0.4 vol% BDDE had pore diams. ranging from a few microns to around 100 μm with a high degree of interconnectivity. The feasibility of this HAG, as an injectable scaffold, to regenerate cartilage and dentin-pulp complex was then demonstrated using a preliminary s.c. microenvironment. The current study could be a ref. to take account of how a crosslinked HA gel should be chosen for specific tissue regeneration.
- 11Collins, M. N.; Birkinshaw, C. Hyaluronic Acid Based Scaffolds for Tissue Engineering - A Review. Carbohydr. Polym. 2013, 92, 1262– 1279, DOI: 10.1016/j.carbpol.2012.10.028Google Scholar11https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXisVSms7c%253D&md5=842c3465f2505666f3c814430e7539ffHyaluronic acid based scaffolds for tissue engineering-A reviewCollins, Maurice N.; Birkinshaw, ColinCarbohydrate Polymers (2013), 92 (2), 1262-1279CODEN: CAPOD8; ISSN:0144-8617. (Elsevier Ltd.)This review focuses on hyaluronic acid (HA) tissue scaffolding materials. Scaffolds are defined in terms of formation mechanisms and mode of action. Soln. properties are discussed as an understanding of the hydrodynamics of HA is fundamental in optimizing the subsequent modification and the chemistries behind important tissue engineering applications that are emerging from recent research on this increasingly valuable carbohydrate polymer are described. Key scaffold characteristics such as mech., biol. function and degrdn. are discussed. The latest technologies behind scaffold processing are assessed and the applications of HA based scaffolds are discussed.
- 12Khunmanee, S.; Jeong, Y.; Park, H. Crosslinking Method of Hyaluronic-Based Hydrogel for Biomedical Applications. J. Tissue Eng. 2017, 8, 204173141772646 DOI: 10.1177/2041731417726464Google ScholarThere is no corresponding record for this reference.
- 13Seif-Naraghi, S. B.; Horn, D.; Schup-Magoffin, P. J.; Christman, K. L. Injectable Extracellular Matrix Derived Hydrogel Provides a Platform for Enhanced Retention and Delivery of a Heparin-Binding Growth Factor. Acta Biomater. 2012, 8, 3695– 3703, DOI: 10.1016/j.actbio.2012.06.030Google Scholar13https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38Xht1Kktb%252FO&md5=771332ec946c70753bfdd4e98acd2214Injectable extracellular matrix derived hydrogel provides a platform for enhanced retention and delivery of a heparin-binding growth factorSeif-Naraghi, Sonya B.; Horn, Dinah; Schup-Magoffin, Pamela J.; Christman, Karen L.Acta Biomaterialia (2012), 8 (10), 3695-3703CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Injectable hydrogels derived from the extracellular matrix (ECM) of decellularized tissues have recently emerged as scaffolds for tissue-engineering applications. Here, we introduce the potential for using a decellularized ECM-derived hydrogel for the improved delivery of heparin-binding growth factors. Immobilization of growth factors on a scaffold has been shown to increase their stability and activity. This can be done via chem. crosslinking, covalent bonding, or by incorporating natural or synthetic growth factor-binding domains similar to those found in vivo in sulfated glycosaminoglycans (GAGs). Many decellularized ECM-derived hydrogels retain native sulfated GAGs, and these materials may therefore provide an excellent delivery platform for heparin-binding growth factors. In this study, the sulfated GAG content of an ECM hydrogel derived from decellularized pericardial ECM was confirmed by Fourier transform IR spectroscopy and its ability to bind basic fibroblast growth factor (bFGF) was established. Delivery in the pericardial matrix hydrogel increased retention of bFGF both in vitro and in vivo in ischemic myocardium compared to delivery in collagen. In a rodent infarct model, intramyocardial injection of bFGF in pericardial matrix enhanced neovascularization by approx. 112% compared to delivery in collagen. Importantly, the newly formed vasculature was anastomosed with existing vasculature. Thus, the sulfated GAG content of the decellularized ECM hydrogel provides a platform for incorporation of heparin-binding growth factors for prolonged retention and delivery.
- 14Seo, Y.; Jung, Y.; Kim, S. H. Decellularized Heart ECM Hydrogel Using Supercritical Carbon Dioxide for Improved Angiogenesis. Acta Biomater. 2018, 67, 270– 281, DOI: 10.1016/j.actbio.2017.11.046Google Scholar14https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXitVaksbzO&md5=2ab93efb29e88bbe22dbfe56f54503d8Decellularized heart ECM hydrogel using supercritical carbon dioxide for improved angiogenesisSeo, Yoojin; Jung, Youngmee; Kim, Soo HyunActa Biomaterialia (2018), 67 (), 270-281CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Initial angiogenesis within the first 3days is crit. for healing ischemic diseases such as myocardial infarction. Recently, decellularized extracellular matrix (dECM) has been reported to provide tissue-derived ECM components and can be used as a scaffold for cell delivery for angiogenesis in tissue engineering. Decellularization by various detergents such as sodium dodecyl sulfate (SDS) and triton X-100 can remove the cell nuclei in tissue organs. However, this leads to ECM structure denaturation, decreased presence of various ECM proteins and cytokines, and loss of mech. properties. To overcome these limitations, in this study, we developed a supercrit. carbon dioxide and ethanol co-solvent (scCO2-EtOH) decellularization method, which is a detergent-free system that prevents ECM structure disruption and retains various angiogenic proteins in the heart dECM, and tested on rat heart tissues. The heart tissue was placed into the scCO2 reactor and decellularized at 37°C and 350bar. After scCO2-EtOH treatment, the effects were evaluated by DNA, collagen, and glycosaminoglycan (GAG) quantification and hematoxylin and eosin and immunofluorescence staining to det. the absence of nucleic acids and preservation of heart ECM components. Similar to the native group, the scCO2-EtOH group contained more ECM components such as collagen, GAGs, collagen I, laminin, and fibronectin and angiogenic factors including vascular endothelial growth factor, fibroblast growth factor, and platelet-derived growth factor and others in comparison to the detergent group. In addn., to est. angiogenesis of the dECM hydrogels, the neutralized dECM soln. was injected in a rat s.c. layer (n=6 in each group: collagen, scCO2-EOH, and detergent group), after which the soln. naturally formed gelation in the s.c. layer. After 3days, the gels were harvested and estd. by immunofluorescence staining and the ImageJ program for angiogenesis anal. Consequently, blood vessel formation and d. of vWF and α-SMA in the scCO2-EtOH group were significantly greater than that in the collagen group. Here we suggest that heart-derived decellularized extracellular matrix (dECM) with scCO2-EtOH treatment is a highly promising angiogenic material for healing in ischemic disease. Supercrit. carbon dioxide (scCO2) in a supercrit. phase has low viscosity and high diffusivity between gas and liq. properties and is known to be affordable, non-toxic, and eco-friendly. Therefore, scCO2 extn. technol. has been extensively used in com. and industrial fields. Recently, decellularized extracellular matrix (dECM) was applied to tissue engineering and regenerative medicine as a scaffold, therapeutic material, and bio-ink for 3D printing. Moreover, the general decellularization method using detergents has limitations including eliminating tissue-derived ECM components and disrupting their structures after decellularization. To overcome these limitations, heart tissues were treated with scCO2-EtOH for decellularization, resulting in preserving of tissue due to the various ECM and angiogenic factors derived. In addn., initiation of angiogenesis was highly induced even after 3days of injection.
- 15Bordbar, S.; Bakhshaiesh, N. L.; Khanmohammadi, M.; Sayahpour, F. A.; Alini, M.; Baghaban Eslaminejad, M. Production and Evaluation of Decellularized Extracellular Matrix Hydrogel for Cartilage Regeneration Derived from Knee Cartilage. J. Biomed. Mater. Res., Part A 2020, 108, 938– 946, DOI: 10.1002/jbm.a.36871Google Scholar15https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjvVWksg%253D%253D&md5=8350496b436f9924c53b3b0d0f72a200Production and evaluation of decellularized extracellular matrix hydrogel for cartilage regeneration derived from knee cartilageBordbar, Sima; Lotfi Bakhshaiesh, Nasrin; Khanmohammadi, Mehdi; Sayahpour, Forough Azam; Alini, Mauro; Baghaban Eslaminejad, MohamadrezaJournal of Biomedical Materials Research, Part A (2020), 108 (4), 938-946CODEN: JBMRCH; ISSN:1549-3296. (John Wiley & Sons, Inc.)Cartilage tissue engineering is the interdisciplinary science that will help to improve cartilage afflictions, such as arthrosis, arthritis, or following joints traumatic injuries. In the present work, we developed an injectable hydrogel which derived from decellularized extracellular matrix of sheep cartilage. Successful decellularization was evaluated by measuring the DNA, glycosaminoglycans (GAG), collagen contents, and histol. analyses. There was a minor difference in GAG and collagen contents among natural cartilage and decellularized tissue as well as ultimate hydrogel. Rheol. anal. showed that the temp. and gelation time of prepd. hydrogel were 37°C and between 5 and 7 min, resp. Mech. properties evaluation indicated a storage modulus of 20 kPa. The results show that prepd. hydrogel possessed cell-friendly microenvironment as confirmed via calcein staining and MTT assay. Also, cells were able to proliferate which obsd. by H&E and alcian blue staining. Cell attachment and proliferation at the surface of the decellularized hydrogel was apparent by Scanning Electron Microscope (SEM) images and microphotographs. Furthermore, the cells embedded within the hydrogel were able to differentiate into chondrocyte with limited evidence of hypertrophy and osteogenesis in utilized cells which proved by SOX9, CoL2, ACAN, and also CoL1 and CoL10 gene expression levels. In summary, the results suggest that developed novel injectable hydrogel from decellularized cartilage could be utilized as a promising substrate for cartilage tissue engineering applications.
- 16Wolf, M. T.; Daly, K. A.; Brennan-Pierce, E. P.; Johnson, S. A.; Carruthers, C. A.; D’Amore, A.; Nagarkar, S. P.; Velankar, S. S.; Badylak, S. F. A Hydrogel Derived from Decellularized Dermal Extracellular Matrix. Biomaterials 2012, 33, 7028– 7038, DOI: 10.1016/j.biomaterials.2012.06.051Google Scholar16https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XhtVaktrfE&md5=0a4178709ca54cb1df8d5bfdddd1c2d1A hydrogel derived from decellularized dermal extracellular matrixWolf, Matthew T.; Daly, Kerry A.; Brennan-Pierce, Ellen P.; Johnson, Scott A.; Carruthers, Christopher A.; D'Amore, Antonio; Nagarkar, Shailesh P.; Velankar, Sachin S.; Badylak, Stephen F.Biomaterials (2012), 33 (29), 7028-7038CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)The ECM of mammalian tissues has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. Such scaffolds are prepd. in many forms including sheets, powders, and hydrogels. ECM hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the inherent bioactivity of native matrix. However, material properties of ECM hydrogels and the effect of these properties upon cell behavior are neither well understood nor controlled. The objective of this study was to prep. and det. the structure, mechanics, and the cell response in vitro and in vivo of ECM hydrogels prepd. from decellularized porcine dermis and urinary bladder tissues. Dermal ECM hydrogels were characterized by a more dense fiber architecture and greater mech. integrity than urinary bladder ECM hydrogels, and showed a dose dependent increase in mech. properties with ECM concn. In vitro, dermal ECM hydrogels supported greater C2C12 myoblast fusion, and less fibroblast infiltration and less fibroblast mediated hydrogel contraction than urinary bladder ECM hydrogels. Both hydrogels were rapidly infiltrated by host cells, primarily macrophages, when implanted in a rat abdominal wall defect. Both ECM hydrogels degraded by 35 days in vivo, but UBM hydrogels degraded more quickly, and with greater amts. of myogenesis than dermal ECM. These results show that ECM hydrogel properties can be varied and partially controlled by the scaffold tissue source, and that these properties can markedly affect cell behavior.
- 17Badylak, S. F.; Freytes, D. O.; Gilbert, T. W. Extracellular Matrix as a Biological Scaffold Material: Structure and Function. Acta Biomater. 2009, 5, 1– 13, DOI: 10.1016/j.actbio.2008.09.013Google Scholar17https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXhsVamsbc%253D&md5=fd27594c129e4f996c49f69a233dbffcExtracellular matrix as a biological scaffold material: structure and functionBadylak, Stephen F.; Freytes, Donald O.; Gilbert, Thomas W.Acta Biomaterialia (2009), 5 (1), 1-13CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)A review. Biol. scaffold materials derived from the extracellular matrix (ECM) of intact mammalian tissues have been successfully used in a variety of tissue engineering/regenerative, medicine applications both in preclin. studies and in clin. applications. Although it is recognized that the materials have constructive remodeling properties, the mechanisms by which functional tissue restoration is achieved are not well understood. There is evidence to support essential roles for both the structural and functional characteristics of the biol. scaffold materials. This paper provides an overview of the compn. and structure of selected ECM scaffold materials, the effects of manufg. methods upon the structural properties and resulting mech. behavior of the scaffold materials, and the in vivo degrdn. and remodeling of ECM scaffolds with an emphasis on tissue function.
- 18Xu, J.; Fang, H.; Zheng, S.; Li, L.; Jiao, Z.; Wang, H.; Nie, Y.; Liu, T.; Song, K. A Biological Functional Hybrid Scaffold Based on Decellularized Extracellular Matrix/Gelatin/Chitosan with High Biocompatibility and Antibacterial Activity for Skin Tissue Engineering. Int. J. Biol. Macromol. 2021, 187, 840– 849, DOI: 10.1016/j.ijbiomac.2021.07.162Google Scholar18https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhslGis7%252FF&md5=bdc31c02e4ec3773c784ee626ce31a9eA biological functional hybrid scaffold based on decellularized extracellular matrix/gelatin/chitosan with high biocompatibility and antibacterial activity for skin tissue engineeringXu, Jie; Fang, Huan; Zheng, Shuangshuang; Li, Liying; Jiao, Zeren; Wang, Hong; Nie, Yi; Liu, Tianqing; Song, KedongInternational Journal of Biological Macromolecules (2021), 187 (), 840-849CODEN: IJBMDR; ISSN:0141-8130. (Elsevier B.V.)Nowadays, decellularized extracellular matrix (dECM) has received widespread attention due to its diversity in providing the unique structural and functional components to support cell growth, and finding material with good biocompatibility and anti-infection capability for skin tissue engineering is still a challenge. In this study, a novel dECM/Gel/CS scaffold with appropriate mech. strength, good antibacterial activity and high biocompatibility was prepd. using a one-pot method. The results showed that the immune components such as cells and DNA (about 98.1%) were successfully removed from the porcine skin tissue. The dECM/Gel/CS scaffolds exhibited an interconnected pore structure and had a high porosity (>90%) to promote cell growth. Moreover, the appropriate elastic modulus (≥482.17 kPa) and degradability (≥80.04% for 15 days) of the scaffolds offered stout "houses" for cell proliferation and suitable degrdn. rate to match the new tissue formation in skin tissue engineering. Furthermore, the addn. of chitosan endowed the scaffold with good antibacterial activity, water and protein absorption capacity to avoid wound infection, and maintain the moisture and nutrition balance. In vitro cytocompatibility studies showed that the presence of dECM effectively enhanced the cell proliferation. Overall, the advanced dECM/Gel/CS scaffold has considerable potential to be applied in skin tissue engineering.
- 19Su, Z.; Ma, H.; Wu, Z.; Zeng, H.; Li, Z.; Wang, Y.; Liu, G.; Xu, B.; Lin, Y.; Zhang, P.; Wei, X. Enhancement of Skin Wound Healing with Decellularized Scaffolds Loaded with Hyaluronic Acid and Epidermal Growth Factor. Mater. Sci. Eng. C 2014, 44, 440– 448, DOI: 10.1016/j.msec.2014.07.039Google Scholar19https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhs1KgtrzI&md5=416018c803a3f51d58443edf981f96bbEnhancement of skin wound healing with decellularized scaffolds loaded with hyaluronic acid and epidermal growth factorSu, Zhongchun; Ma, Huan; Wu, Zhengzheng; Zeng, Huilan; Li, Zhizhong; Wang, Yuechun; Liu, Gexiu; Xu, Bin; Lin, Yongliang; Zhang, Peng; Wei, XingMaterials Science & Engineering, C: Materials for Biological Applications (2014), 44 (), 440-448CODEN: MSCEEE; ISSN:0928-4931. (Elsevier B.V.)Current therapy for skin wound healing still relies on skin transplantation. Many studies were done to try to find out ways to replace skin transplantation, but there is still no effective alternative therapy. In this study, decellularized scaffolds were prepd. from pig peritoneum by a series of phys. and chem. treatments, and scaffolds loaded with hyaluronic acid (HA) and epidermal growth factor (EGF) were tested for their effect on wound healing. MTT assay showed that EGF increased NIH3T3 cell viability and confirmed that EGF used in this study was biol. active in vitro. Scanning electron microscope (SEM) showed that HA stably attached to scaffolds even after soaking in PBS for 48 h. ELISA assay showed that HA increased the adsorption of EGF to scaffolds and sustained the release of EGF from scaffolds. Animal study showed that the wounds covered with scaffolds contg. HA and EGF recovered best among all 4 groups and had wound healing rates of 49.86%, 70.94% and 87.41% resp. for days 10, 15 and 20 post-surgery compared to scaffolds alone with wound healing rates of 29.26%, 42.80% and 70.14%. In addn., the wounds covered with scaffolds contg. EGF alone were smaller than no EGF scaffolds on days 10, 15 and 20 post-surgery. Hematoxylin-Eosin (HE) staining confirmed these results by showing that on days 10, 15 and 20 post-surgery, the thicker epidermis and dermis layers were obsd. in the wounds covered with scaffolds contg. HA and EGF than scaffolds alone. In addn., the thicker epidermis and dermis layers were also obsd. in the wounds covered with scaffolds contg. EGF than scaffolds alone. Skin appendages were obsd. on day 20 only in the wound covered with scaffolds contg. HA and EGF. These results demonstrate that the scaffolds contg. HA and EGF can enhance wound healing.
- 20Chen, W.; Xu, Y.; Li, Y.; Jia, L.; Mo, X.; Jiang, G.; Zhou, G. 3D Printing Electrospinning Fiber-Reinforced Decellularized Extracellular Matrix for Cartilage Regeneration. Chem. Eng. J. 2020, 382, 122986 DOI: 10.1016/j.cej.2019.122986Google Scholar20https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvFejtrzL&md5=1a35b80756b9832a8a258796ab71f0d43D printing electrospinning fiber-reinforced decellularized extracellular matrix for cartilage regenerationChen, Weiming; Xu, Yong; Li, Yaqiang; Jia, Litao; Mo, Xiumei; Jiang, Gening; Zhou, GuangdongChemical Engineering Journal (Amsterdam, Netherlands) (2020), 382 (), 122986CODEN: CMEJAJ; ISSN:1385-8947. (Elsevier B.V.)Cartilage decellularized matrix (CDM) is considered a promising biomaterial for fabricating cartilage tissue engineering scaffolds. An ideal CDM-based scaffold should possess customizable 3D shape for complex tissue regeneration and proper pore size for cell infiltration, as well as provide mech. support for cell growth. 3D printing is an efficiently technique for prepg. customizable 3D scaffolds, however, fabricating CDM-based 3D-printed scaffolds with customizable shapes, proper pore structure and satisfactory mech. properties remains a challenge. In the current study, to achieve customizable CDM-based 3D scaffolds, CDM was successfully processed into inks suitable for 3D printing. Further, the poor mechanics of CDM-based scaffolds were significantly improved by adding electrospinning fiber into the CDM-based inks for 3D printing. Importantly, the 3D-printed electrospinning fiber-reinforced CDM-based scaffold presented good biocompatibility and can enhance repair articular cartilage defects in rabbits. The current study provides a novel strategy for printing electrospinning fiber-reinforced CDM-based scaffolds for tissue regeneration.
- 21Wang, Z.; Li, Z.; Li, Z.; Wu, B.; Liu, Y.; Wu, W. Cartilaginous Extracellular Matrix Derived from Decellularized Chondrocyte Sheets for the Reconstruction of Osteochondral Defects in Rabbits. Acta Biomater. 2018, 81, 129– 145, DOI: 10.1016/j.actbio.2018.10.005Google Scholar21https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhvFSqu7jK&md5=7b2448e86db36a983a1cf4c124800d75Cartilaginous extracellular matrix derived from decellularized chondrocyte sheets for the reconstruction of osteochondral defects in rabbitsWang, Zhifa; Li, Zhiye; Li, Zhijin; Wu, Buling; Liu, Yanpu; Wu, WeiActa Biomaterialia (2018), 81 (), 129-145CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Cartilaginous extracellular matrix (ECM) materials derived from decellularized native articular cartilage are widely used in cartilage regeneration. However, it is difficult for endogenous cells to migrate into ECM derived from native cartilage owing to its nonporous structure and dense nature. We cultured chondrocytes harvested from the auricular cartilage of 4-wk-old New Zealand rabbits and enabled them to form cell sheets. In vitro microstructural examn. and mech. tests demonstrated that 1% SDS not only removed chondrocytes completely but also maintained the native architecture and compn. of ECM, thus avoiding the use of high-concn. SDS. In conclusion, our results suggested that the chondrocyte sheets decellularized with 1% SDS preserved the integrity and bioactivity, which favored cell recruitment and enabled osteochondral regeneration in the knee joints of rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation. Our results suggested that cartilaginous ECM favored cell recruitment and enabled osteochondral regeneration in rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation. SDS 1% adequately decellularized the chondrocytes in cell sheets, whereas it maintained the native architecture and compn. of ECM, thereby avoiding the use of high-concn. SDS and providing a new way to acquire cartilaginous ECM.
- 22Han, T. T. Y.; Toutounji, S.; Amsden, B. G.; Flynn, L. E. Adipose-Derived Stromal Cells Mediate in Vivo Adipogenesis, Angiogenesis and Inflammation in Decellularized Adipose Tissue Bioscaffolds. Biomaterials 2015, 72, 125– 137, DOI: 10.1016/j.biomaterials.2015.08.053Google Scholar22https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXhsVaqsL7N&md5=6a74a992d2da07c5e4a667862c9a6f9dAdipose-derived stromal cells mediate in vivo adipogenesis, angiogenesis and inflammation in decellularized adipose tissue bioscaffoldsHan, Tim Tian Y.; Toutounji, Sandra; Amsden, Brian G.; Flynn, Lauren E.Biomaterials (2015), 72 (), 125-137CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)Decellularized adipose tissue (DAT) has shown promise as an adipogenic bioscaffold for soft tissue augmentation and reconstruction. The objective of the current study was to investigate the effects of allogeneic adipose-derived stem/stromal cells (ASCs) on in vivo fat regeneration in DAT bioscaffolds using an immunocompetent rat model. ASC seeding significantly enhanced angiogenesis and adipogenesis, with cell tracking studies indicating that the newly-forming tissues were host-derived. Incorporating ASCs also mediated the inflammatory response and promoted a more constructive macrophage phenotype. A fraction of the CD163+ macrophages in the implants expressed adipogenic markers, with higher levels of this "adipocyte-like" phenotype in proximity to the developing adipose tissues. Our results indicate that the combination of ASCs and adipose extracellular matrix (ECM) provides an inductive microenvironment for adipose regeneration mediated by infiltrating host cell populations. The DAT scaffolds are a useful tissue-specific model system for investigating the mechanisms of in vivo adipogenesis that may help to develop a better understanding of this complex process in the context of both regeneration and disease. Overall, combining adipose-derived matrixes with ASCs is a highly promising approach for the in situ regeneration of host-derived adipose tissue.
- 23Han, W.; Singh, N. K.; Kim, J. J.; Kim, H.; Kim, B. S.; Park, J. Y.; Jang, J.; Cho, D. W. Directed Differential Behaviors of Multipotent Adult Stem Cells from Decellularized Tissue/Organ Extracellular Matrix Bioinks. Biomaterials 2019, 224, 119496 DOI: 10.1016/j.biomaterials.2019.119496Google Scholar23https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvVamt7fO&md5=a3eb11fd9fa07b402b08429ec61ef19aDirected differential behaviors of multipotent adult stem cells from decellularized tissue/organ extracellular matrix bioinksHan, Wonil; Singh, Narendra K.; Kim, Joeng Ju; Kim, Hyeonji; Kim, Byoung Soo; Park, Ju Young; Jang, Jinah; Cho, Dong-WooBiomaterials (2019), 224 (), 119496CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)The decellularized tissue/organ extracellular matrix (dECM) is a naturally derived biomaterial that inherits various functional components from the native tissue or organ. Recently, various kinds of tissue/organ dECM bioinks capable of encapsulating cells, combined with 3D cell printing, have enabled remarkable progress in tissue engineering and regenerative medicine. However, the way in which the dECM component compns. of each tissue of different origins interact with cells and dictate tissue-specific cell behavior in the 3D microenvironment remains mostly unknown. To address this issue, in-depth differential proteomic analyses of four porcine dECMs were performed. Specifically, the differential variations of matrisome protein compn. in each decellularized tissue type were also uncovered, which can play a significant role by affecting the resident cells in specific tissues. Furthermore, microarray analyses of human bone marrow mesenchymal stem cells (hBMMSCs) printed with various dECM bioinks were conducted to reveal the effect of compositional variations in a tissue-specific manner at the cellular level depending on the multipotency of MSCs. Through whole transcriptome anal., differential expression patterns of genes were obsd. in a tissue-specific manner, and this research provides strong evidence of the tissue-specific functionalities of dECM bioinks.
- 24Zhou, H.; Chen, R.; Wang, J.; Lu, J.; Yu, T.; Wu, X.; Xu, S.; Li, Z.; Jie, C.; Cao, R.; Yang, Y.; Li, Y.; Meng, D. Biphasic Fish Collagen Scaffold for Osteochondral Regeneration. Mater. Des. 2020, 195, 108947 DOI: 10.1016/j.matdes.2020.108947Google Scholar24https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhsF2qsbzM&md5=5107bf9dd64bd9cb3aa0ba8429ecbb88Biphasic fish collagen scaffold for osteochondral regenerationZhou, Haichao; Chen, Ru; Wang, Jinpeng; Lu, Jia; Yu, Tao; Wu, Xinbo; Xu, Shaochen; Li, Zihua; Jie, Chen; Cao, Runfeng; Yang, Yunfeng; Li, Yaqiang; Meng, DepengMaterials & Design (2020), 195 (), 108947CODEN: MADSD2; ISSN:0264-1275. (Elsevier Ltd.)Developing a biphasic scaffold that can concurrently regenerate both cartilage and bone of osteochondral defects (OCDs) is a challenge. Fish collagen (FC) is currently considered an alternative scaffold to mammalian collagen (MC) due to its safety, accessibility, lower price, and similar biol. properties compared to those of MC. Here, we developed bilayer FC-based composite scaffolds with different components and pore sizes to modulate the differentiation fate of bone marrow stem cells (BMSCs): chondroitin sulfate-incorporated FC scaffolds (FC-CS) with small pores (approx. 128μm) as the top layer and hydroxyapatite-incorporated FC scaffolds (FC-HA) with larger pores (approx. 326μm) as the bottom layer. Both the FC-CS and FC-HA scaffolds possessed good cytocompatibility, excellent water absorption, suitable biodegradability and high cell seeding efficiency. The in vitro results indicated that FC-CS and FC-HA promote chondrogenesis and osteogenesis of BMSCs, resp., as validated by gene expression and histol. examn. Furthermore, compared to the empty group in a rabbit OCD model, the bilayer scaffold significantly induced simultaneous regeneration of cartilage and subchondral bone after 6 and 12 wk of implantation, which was confirmed by gross, histol., and microcomputed tomog. images. Our findings demonstrated that the FC-based bilayer scaffold is a promising scaffold for the repair of OCD.
- 25Kirsner, R. S.; Margolis, D. J.; Baldursson, B. T.; Petursdottir, K.; Davidsson, O. B.; Weir, D.; Lantis, J. C. Fish Skin Grafts Compared to Human Amnion/Chorion Membrane Allografts: A Double-Blind, Prospective, Randomized Clinical Trial of Acute Wound Healing. Wound Repair Regener. 2020, 28, 75– 80, DOI: 10.1111/wrr.12761Google Scholar25https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3Mrns1ShsA%253D%253D&md5=90d00ef9004c0d03ed7e30810307702dFish skin grafts compared to human amnion/chorion membrane allografts: A double-blind, prospective, randomized clinical trial of acute wound healingKirsner Robert S; Margolis David J; Margolis David J; Baldursson Baldur T; Petursdottir Kristin; Davidsson Olafur B; Weir Dot; Lantis John C 2ndWound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society (2020), 28 (1), 75-80 ISSN:.Chronic, nonhealing wounds consume a great deal of healthcare resources and are a major public health problem, associated with high morbidity and significant economic costs. Skin grafts are commonly used to facilitate wound closure. The grafts can come from the patient's own skin (autograft), a human donor (allograft), or from a different species (xenograft). A fish skin xenograft from cold-water fish (Atlantic cod, Gadus morhua) is a relatively recent option that shows promising preclinical and clinical results in wound healing. Chronic wounds vary greatly in etiology and nature, requiring large cohorts for effective comparison between therapeutic alternatives. In this study, we attempted to imitate the status of a freshly debrided chronic wound by creating acute full-thickness wounds, 4 mm in diameter, on healthy volunteers to compare two materials frequently used to treat chronic wounds: fish skin and dHACM. The purpose is to give an indication of the efficacy of the two therapeutic alternatives in the treatment of chronic wounds in a simple, standardized, randomized, controlled, double-blind study. All volunteers were given two identical punch biopsy wounds, one of which was treated with a fish skin graft and the other with dehydrated human amnion/chorion membrane allograft (dHACM). In the study, 170 wounds were treated (85 wounds per group). The primary endpoint was defined as time to heal (full epithelialization) by blinded assessment at days 14, 18, 21, 25, and 28. The superiority hypothesis was that the fish skin grafts would heal the wounds faster than the dHACM. To evaluate the superiority hypothesis, a mixed Cox proportional hazard model was used. Wounds treated with fish skin healed significantly faster (hazard ratio 2.37; 95% confidence interval: (1.75-3.22; p = 0.0014) compared with wounds treated with dHACM. The results show that acute biopsy wounds treated with fish skin grafts heal faster than wounds treated with dHACM.
- 26Govindharaj, M.; Roopavath, U. K.; Rath, S. N. Valorization of Discarded Marine Eel Fish Skin for Collagen Extraction as a 3D Printable Blue Biomaterial for Tissue Engineering. J. Cleaner Prod. 2019, 230, 412– 419, DOI: 10.1016/j.jclepro.2019.05.082Google Scholar26https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtVKgurvP&md5=057db7abdea5b3a139fab2790428ea84Valorization of discarded Marine Eel fish skin for collagen extraction as a 3D printable blue biomaterial for tissue engineeringGovindharaj, Mano; Roopavath, Uday Kiran; Rath, Subha NarayanJournal of Cleaner Production (2019), 230 (), 412-419CODEN: JCROE8; ISSN:0959-6526. (Elsevier Ltd.)Discarded marine Eel fish skin has essential properties of biomaterials for potential use in tissue engineering application. Processing and prepn. of eel fish for edible purpose requires the removal of skin due to its thick size, which is treated as a waste. A huge amt. of Eel skin is dumped as a waste material which leads to marine environmental pollution. To overcome this issue, we have isolated collagen from the discarded marine Eel skin as a potential blue biomaterial. Further, the isolated collagen was incorporated into alginate hydrogel to fabricate scaffolds using extrusion-based 3D printing technol. Swelling, degrdn. and biocompatibility were evaluated for lyophilized scaffolds. Biocompatibility studies were performed on hUMSCs (Human Umbilical cord Derived Mesenchymal Stem Cells) by live/dead staining using FDA (fluorescein diacetate)/PI (Propidium Iodide). The quant. evaluation of metabolic activity was performed using Alamar Blue (AB) dye redn. assay. All the hydrogels with collagen show enhanced metabolic activity and cell proliferation compared to alginate hydrogels without collagen. The utilization of Eel skin derived collagen for 3D printing application was not yet reported. Moreover, sustainable utilization of renewable marine Eel skin discard as a novel blue biomaterial is of immense value due to its low cost and has great potential for further tissue engineering applications.
- 27Blanco, M.; Vázquez, J. A.; Pérez-Martín, R. I.; Sotelo, C. G. Collagen Extraction Optimization from the Skin of the Small-Spotted Catshark (S. Canicula) by Response Surface Methodology. Mar. Drugs 2019, 17, 40 DOI: 10.3390/md17010040Google Scholar27https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtFGmtrzJ&md5=76da1a9f390bb5c4fba379151e0dcf2fCollagen extraction optimization from the skin of the small-spotted catshark (S. canicula) by response surface methodologyBlanco, Maria; Vazquez, Jose Antonio; Perez-Martin, Ricardo I.; Sotelo, Carmen G.Marine Drugs (2019), 17 (1), 40CODEN: MDARE6; ISSN:1660-3397. (MDPI AG)The small-spotted catshark is one of the most abundant elasmobranchs in the Northeastern Atlantic Ocean. Although its landings are devoted for human consumption, in general this species has low com. value with high discard rates, reaching 100% in some European fisheries. The eduction of post-harvest losses (discards and byproducts) by promotion of a full use of fishing captures is one of the main goals of EU fishing policies. As marine collagens are increasingly used as alternatives to mammalian collagens for cosmetics, tissue engineering, etc., fish skins represent an excellent and abundant source for obtaining this biomol. The aim of this study was to analyze the influence of chem. treatment concn., temp. and time on the extractability of skin collagen from this species. Two exptl. designs, one for each of the main stages of the process, were performed by means of Response Surface Methodol. (RSM). The combined effect of NaOH concn., time and temp. on the amt. of collagen recovered in the first stage of the collagen extn. procedure was studied. Then, skins treated under optimal NaOH conditions were subjected to a second exptl. design, to study the combined effect of AcOH concn., time and temp. on the collagen recovery by means of yield, amino acid content and SDS-PAGE characterization. Values of independent variables maximizing collagen recovery were 4°C, 2 h and 0.1 M NaOH (pre-treatment) and 25°C, 34 h and 1 M AcOH (collagen extn.).
- 28Lim, Y.; Ok, Y.-J.; Hwang, S.; Kwak, J.; Yoon, S. Marine Collagen as A Promising Biomaterial For. Mar. Drugs 2019, 17, 467 DOI: 10.3390/md17080467Google Scholar28https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXks1Oks7g%253D&md5=e9dd3e6d1288e11100187bd1ec534c35Marine collagen as a promising biomaterial for biomedical applicationsLim, Ye-Seon; Ok, Ye-Jin; Hwang, Seon-Yeong; Kwak, Jong-Young; Yoon, SikMarine Drugs (2019), 17 (8), 467CODEN: MDARE6; ISSN:1660-3397. (MDPI AG)This review focuses on the expanding role of marine collagen (MC)-based scaffolds for biomedical applications. A scaffold-a three-dimensional (3D) structure fabricated from biomaterials-is a key supporting element for cell attachment, growth, and maintenance in 3D cell culture and tissue engineering. The mech. and biol. properties of the scaffolds influence cell morphol., behavior, and function. MC, collagen derived from marine organisms, offers advantages over mammalian collagen due to its biocompatibility, biodegradability, easy extractability, water soly., safety, low immunogenicity, and low prodn. costs. In recent years, the use of MC as an increasingly valuable scaffold biomaterial has drawn considerable attention from biomedical researchers. The characteristics, isolation, phys., and biochem. properties of MC are discussed as an understanding of MC in optimizing the subsequent modification and the chemistries behind important tissue engineering applications. The latest technologies behind scaffold processing are assessed and the biomedical applications of MC and MC-based scaffolds, including tissue engineering and regeneration, wound dressing, drug delivery, and therapeutic approach for diseases, esp. those assocd. with metabolic disturbances such as obesity and diabetes, are discussed. Despite all the challenges, MC holds great promise as a biomaterial for developing medical products and therapeutics.
- 29Girardeau-Hubert, S.; Lynch, B.; Zuttion, F.; Label, R.; Rayee, C.; Brizion, S.; Ricois, S.; Martinez, A.; Park, E.; Kim, C.; Marinho, P. A.; Shim, J. H.; Jin, S.; Rielland, M.; Soeur, J. Impact of Microstructure on Cell Behavior and Tissue Mechanics in Collagen and Dermal Decellularized Extra-Cellular Matrices. Acta Biomater. 2022, 143, 100– 114, DOI: 10.1016/j.actbio.2022.02.035Google Scholar29https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XhsVSgtLnL&md5=3f9072612ae4d39007df8771f8b46986Impact of microstructure on cell behavior and tissue mechanics in collagen and dermal decellularized extra-cellular matricesGirardeau-Hubert, Sarah; Lynch, Barbara; Zuttion, Francesca; Label, Rabab; Rayee, Chrystelle; Brizion, Sebastien; Ricois, Sylvie; Martinez, Anthony; Park, Eunhye; Kim, Changhwan; Marinho, Paulo Andre; Shim, Jin-Hyung; Jin, Songwan; Rielland, Maite; Soeur, JeremieActa Biomaterialia (2022), 143 (), 100-114CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Skin models are used for many applications such as research and development or grafting. Unfortunately, most lack a proper microenvironment producing poor mech. properties and inaccurate extra-cellular matrix compn. and organization. In this report we focused on mech. properties, extra-cellular matrix organization and cell interactions in human skin samples reconstructed with pure collagen or dermal decellularized extra-cellular matrixes (S-dECM) and compared them to native human skin. We found that Full-thickness S-dECM samples presented stiffness two times higher than collagen gel and similar to ex vivo human skin, and proved for the first time that keratinocytes also impact dermal mech. properties. This was correlated with larger fibers in S-dECM matrixes compared to collagen samples and with a differential expression of F-actin, vinculin and tenascin C between S-dECM and collagen samples. This is clear proof of the microenvironments impact on cell behaviors and mech. properties. In vitro skin models have been used for a long time for clin. applications or in vitro knowledge and evaluation studies. However, most lack a proper microenvironment producing a poor combination of mech. properties and appropriate biol. outcomes, partly due to inaccurate extra-cellular matrix (ECM) compn. and organization. This can lead to limited predictivity and weakness of skin substitutes after grafting. This study shows, for the first time, the importance of a complex and rich microenvironment on cell behaviors, matrix macro- and micro-organization and mech. properties. The increased compn. and organization complexity of dermal skin decellularized extra-cellular matrix populated with differentiated cells produces in vitro skin models closer to native human skin physiol.
- 30Chameettachal, S.; Sasikumar, S.; Sethi, S.; Sriya, Y.; Pati, F. Tissue/Organ-Derived Bioink Formulation for 3D Bioprinting. J. 3D Print. Med. 2019, 3, 39– 54, DOI: 10.2217/3dp-2018-0024Google Scholar30https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXkslamt7Y%253D&md5=c70ce9cf2f9c97eaf43066f5b77ec00dTissue/organ-derived bioink formulation for 3D bioprintingChameettachal, Shibu; Sasikumar, Shyama; Sethi, Soumya; Sriya, Yeleswarapu; Pati, FalguniJournal of 3D Printing in Medicine (2019), 3 (1), 39-54CODEN: JDPMAT; ISSN:2059-4763. (Future Medicine Ltd.)Tissue/organ-derived bioink formulations open up new avenues in 3D bioprinting research with the potential to create functional tissue or organs. Printing of tissue construct largely depends on material properties, as it needs to be fabricated in an aq. environment while encapsulating living cells. The decellularized extracellular matrix bioinks proved to be a potential option for functional tissue development in vivo and as an alternative to chem. cross-linked bioinks. However, certain limitations such as printability and limited mech. strength need to be addressed for enhancing their widespread applications. By drawing knowledge from the existing literature, emphasis has been given in this review to the development of decellularized extracellular matrix bioinks and their applications in printing functional tissue constructs.
- 31Reing, J. E.; Brown, B. N.; Daly, K. A.; Freund, J. M.; Gilbert, T. W.; Hsiong, S. X.; Huber, A.; Kullas, K. E.; Tottey, S.; Wolf, M. T.; Badylak, S. F. The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds. Biomaterials 2010, 31, 8626– 8633, DOI: 10.1016/j.biomaterials.2010.07.083Google Scholar31https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXht1ehtLnP&md5=5f2f35e650c7a0b64fdfadc67becd2c7The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffoldsReing, Janet E.; Brown, Bryan N.; Daly, Kerry A.; Freund, John M.; Gilbert, Thomas W.; Hsiong, Susan X.; Huber, Alexander; Kullas, Karen E.; Tottey, Stephen; Wolf, Matthew T.; Badylak, Stephen F.Biomaterials (2010), 31 (33), 8626-8633CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)Biol. materials from various species and tissues are commonly used as surgical meshes or scaffolds for tissue reconstruction. Extracellular matrix (ECM) represents the secreted product of the cells comprising each tissue and organ, and therefore provides a unique biol. material for selected regenerative medicine applications. Minimal disruption of ECM ultrastructure and content during tissue processing is typically desirable. The objective of this study was to systematically evaluate effects of commonly used tissue processing steps upon porcine dermal ECM scaffold compn., mech. properties, and cytocompatibility. Processing steps evaluated included liming and hot water sanitation, trypsin/SDS/TritonX-100 decellularization, and trypsin/TritonX-100 decellularization. Liming decreased the growth factor and glycosaminoglycan content, the mech. strength, and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for all). Hot water sanitation treatment decreased only the growth factor content of the ECM (p ≤ 0.05). Trypsin/SDS/TritonX-100 decellularization decreased the growth factor content and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). Trypsin/Triton X-100 decellularization also decreased the growth factor content of the ECM but increased the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). The authors conclude that processing steps evaluated in the present study affect content, mech. strength, and/or cytocompatibility of the resultant porcine dermal ECM, and therefore care must be taken in choosing appropriate processing steps to maintain the beneficial effects of ECM in biol. scaffolds.
- 32Zhang, Q.; Johnson, J. A.; Dunne, L. W.; Chen, Y.; Iyyanki, T.; Wu, Y.; Chang, E. I.; Branch-Brooks, C. D.; Robb, G. L.; Butler, C. E. Decellularized Skin/Adipose Tissue Flap Matrix for Engineering Vascularized Composite Soft Tissue Flaps. Acta Biomater. 2016, 35, 166– 184, DOI: 10.1016/j.actbio.2016.02.017Google Scholar32https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xis1CqtLY%253D&md5=85c5917bc2329157c22a3b88025b0cfdDecellularized skin/adipose tissue flap matrix for engineering vascularized composite soft tissue flapsZhang, Qixu; Johnson, Joshua A.; Dunne, Lina W.; Chen, Youbai; Iyyanki, Tejaswi; Wu, Yewen; Chang, Edward I.; Branch-Brooks, Cynthia D.; Robb, Geoffrey L.; Butler, Charles E.Acta Biomaterialia (2016), 35 (), 166-184CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Using a perfusion decellularization protocol, we developed a decellularized skin/adipose tissue flap (DSAF) comprising extracellular matrix (ECM) and intact vasculature. Our DSAF had a dominant vascular pedicle, microcirculatory vascularity, and a sensory nerve network and retained three-dimensional (3D) nanofibrous structures well. DSAF, which was composed of collagen and laminin with well-preserved growth factors (e.g., vascular endothelial growth factor, basic fibroblast growth factor), was successfully repopulated with human adipose-derived stem cells (hASCs) and human umbilical vein endothelial cells (HUVECs), which integrated with DSAF and formed 3D aggregates and vessel-like structures in vitro. We used microsurgery techniques to re-anastomose the recellularized DSAF into nude rats. In vivo, the engineered flap construct underwent neovascularization and constructive remodeling, which was characterized by the predominant infiltration of M2 macrophages and significant adipose tissue formation at 3 mo postoperatively. Our results indicate that DSAF co-cultured with hASCs and HUVECs is a promising platform for vascularized soft tissue flap engineering. This platform is not limited by the flap size, as the entire construct can be immediately perfused by the recellularized vascular network following simple re-integration into the host using conventional microsurgical techniques. Significant soft tissue loss resulting from traumatic injury or tumor resection often requires surgical reconstruction using autologous soft tissue flaps. However, the limited availability of qual. autologous flaps as well as the donor site morbidity significantly limits this approach. Engineered soft tissue flap grafts may offer a clin. relevant alternative to the autologous flap tissue. In this study, we engineered vascularized soft tissue free flap by using skin/adipose flap extracellular matrix scaffold (DSAF) in combination with multiple types of human cells. Following vascular reanastomosis in the recipient site, the engineered products successful regenerated large-scale fat tissue in vivo. This approach may provide a translatable platform for composite soft tissue free flap engineering for microsurgical reconstruction.
- 33Liu, D.; Wei, G.; Li, T.; Hu, J.; Lu, N.; Regenstein, J. M.; Zhou, P. Effects of Alkaline Pretreatments and Acid Extraction Conditions on the Acid-Soluble Collagen from Grass Carp (Ctenopharyngodon Idella) Skin. Food Chem. 2015, 172, 836– 843, DOI: 10.1016/j.foodchem.2014.09.147Google Scholar33https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhslajtb7L&md5=423b7904b4e4a78a130acb3f00793958Effects of alkaline pretreatments and acid extraction conditions on the acid-soluble collagen from grass carp (Ctenopharyngodon idella) skinLiu, Dasong; Wei, Guanmian; Li, Tiancheng; Hu, Jinhua; Lu, Naiyan; Regenstein, Joe M.; Zhou, PengFood Chemistry (2015), 172 (), 836-843CODEN: FOCHDJ; ISSN:0308-8146. (Elsevier Ltd.)This study investigated the effects of alk. pretreatments and acid extn. conditions on the prodn. of acid-sol. collagen (ASC) from grass carp skin. For alk. pretreatment, 0.05 and 0.1 M NaOH removed non-collagenous proteins without significant loss of ASC at 4, 10, 15 and 20 °C; while 0.2 and 0.5 M NaOH caused significant loss of ASC, and 0.5 M NaOH caused structural modification of ASC at 15 and 20 °C. For acid extn. at 4, 10, 15 and 20 °C, ASC was partly extd. by 0.1 and 0.2 M acetic acid, while 0.5 and 1.0 M acetic acid resulted in almost complete extn. The processing conditions involving 0.05-0.1 M NaOH for pretreatment, 0.5 M acetic acid for extn. and 4-20 °C for both pretreatment and extn., produced ASC with the structural integrity being well maintained and hence were recommended to prep. ASC from grass carp skin in practical application.
- 34Meng, D.; Tanaka, H.; Kobayashi, T.; Hatayama, H.; Zhang, X.; Ura, K.; Yunoki, S.; Takagi, Y. The Effect of Alkaline Pretreatment on the Biochemical Characteristics and Fibril-Forming Abilities of Types I and II Collagen Extracted from Bester Sturgeon by-Products. Int. J. Biol. Macromol. 2019, 131, 572– 580, DOI: 10.1016/j.ijbiomac.2019.03.091Google Scholar34https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXlvVWht7s%253D&md5=41b12c2b9764573c1ddc31372f3922aeThe effect of alkaline pretreatment on the biochemical characteristics and fibril-forming abilities of types I and II collagen extracted from bester sturgeon by-productsMeng, Dawei; Tanaka, Hiroyuki; Kobayashi, Taishi; Hatayama, Hirosuke; Zhang, Xi; Ura, Kazuhiro; Yunoki, Shunji; Takagi, YasuakiInternational Journal of Biological Macromolecules (2019), 131 (), 572-580CODEN: IJBMDR; ISSN:0141-8130. (Elsevier B.V.)Non-mammalian collagens have attracted increasing attention for industrial and biomedical use. We have therefore evaluated extn. conditions and the biochem. properties of collagens from aquacultured sturgeon. Pepsin-sol. type I and type II collagen were resp. extd. from the skin and notochord of bester sturgeon byproducts, with yields of 63.9 ± 0.19% and 35.5 ± 0.68%. Collagen extn. efficiency was improved by an alk. pretreatment of the skin and notochord (fewer extn. cycles were required), but the final yields decreased to 56.2 ± 0.84% for type I and 31.8 ± 1.13% for type II. Alk. pretreatment did not affect the thermal stability or triple-helical structure of both types of collagen. Types I and II collagen formed re-assembled fibril structures in vitro, under different conditions. Alk. pretreatment slowed down the formation of type I collagen fibrils and specifically inhibited the formation of thick fibril-bundle structures. In contrast, alk. pretreatment did not change type II collagen fibril formation. In conclusion, alk. pretreatment of sturgeon skin and notochord is an effective method to accelerate collagen extn. process of types I and II collagen without changing their biochem. properties. However, it decreases the yield of both collagens and specifically changes the fibril-forming ability of type I collagen.
- 35Furtado, M.; Chen, L.; Chen, Z.; Chen, A.; Cui, W. Development of Fish Collagen in Tissue Regeneration and Drug Delivery. Eng. Regen. 2022, 3, 217– 231, DOI: 10.1016/j.engreg.2022.05.002Google ScholarThere is no corresponding record for this reference.
- 36Yoshimura, K.; Terashima, M.; Hozan, D.; Shirai, K. Preparation and Dynamic Viscoelasticity Characterization of Alkali- Solubilized Collagen from Shark Skin. J. Agric. Food Chem. 2000, 48, 685– 690, DOI: 10.1021/jf990389dGoogle Scholar36https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXhtlKjtrw%253D&md5=d138b61787c55e1d13b675ed31412e73Preparation and Dynamic Viscoelasticity Characterization of Alkali-Solubilized Collagen from Shark SkinYoshimura, Keiji; Terashima, Mariko; Hozan, Daiki; Shirai, KunioJournal of Agricultural and Food Chemistry (2000), 48 (3), 685-690CODEN: JAFCAU; ISSN:0021-8561. (American Chemical Society)Alkali-solubilized collagens, prepd. by alkali-acid extn. and alkali direct extn. (abbreviated AASC and ALSC, resp.), were characterized by dynamic viscoelastic measurement of collagen soln. (10 mg/mL). The optimum preparative conditions in terms of yield and polypeptide size are as follows: for the alkali-acid extn., a pretreatment with 0.5 or 1 M NaOH contg. 15% Na2SO4 within 5 days at 20 °C followed by the subsequent acid extn., and for the alk. direct extn., a treatment with 0.5 M NaOH contg. 10% NaCl at 4 °C for 20-30 days. A major portion of the polypeptide sizes of AASC and ALSC is composed of α chains (α1 and α2). Dynamic viscoelasticity of collagen soln. was measured as a function of temp. AASC showed a greater contribution of elastic behavior rather than viscous behavior. On the contrary, ALSC exhibits a stronger viscous behavior than elastic behavior.
- 37Bowes, J. H.; Kenten, R. H. The Effect of Alkalis on Collagen. Biochem. J. 1948, 43, 365– 372, DOI: 10.1042/bj0430365Google Scholar37https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaH1MXitVSqtA%253D%253D&md5=163920312283531adbf4cb3b9cdf492dEffect of alkalies on collagenBowes, Joane H.; Kenten, R. H.Biochemical Journal (1948), 43 (), 365-72CODEN: BIJOAK; ISSN:0264-6021.Carefully prepd. and chemically unaltered collagen treated with alkali at pH 13 hydrolyzes off amide groups. This is the main reaction taking place. A small portion of guanidine groups is converted to ornithine and urea, and even a smaller portion to citrulline and NH3. Probably some peptide links of proline and hydroxyproline are broken. About 5% of the material becomes solubilized. Following alkali treatment (Ca(OH)2 + NaOH) the collagen swells more than before.
- 38Hattori, S.; Adachi, E.; Ebihara, T.; Shirai, T.; Someki, I.; Irie, S. Alkali-Treated the Ligand Collagen Activity Retained the the Adhesion Triple via Conformation Integrin for The Method of Extracting Collagen from Connective Tissue with a Solution of Sodium Hydroxide and Monomethylamine Was Developed about 30 Years Ago (. J. Biochem. 1999, 125, 676– 684, DOI: 10.1093/oxfordjournals.jbchem.a022336Google Scholar38https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXjsVyrtL4%253D&md5=9126263c0e61c88df79b4574d6bba37fAlkali-treated collagen retained the triple helical conformation and the ligand activity for the cell adhesion via α2β1 integrinHattori, Shunji; Adachi, Eijiro; Ebihara, Tetsuya; Shirai, Tomoko; Someki, Iori; Irie, ShinkichiJournal of Biochemistry (1999), 125 (4), 676-684CODEN: JOBIAO; ISSN:0021-924X. (Japanese Biochemical Society)Alk. treatment is a good method for extg. collagen with high recovery even from an aged animal specimen. However, the properties of collagen treated under alk. conditions have not been well established yet. By the treatment with a soln. of 3% sodium hydroxide and 1.9% monomethylamine, the isoelec. point of type I collagen was lowered from 9.3 to 4.8 because of the conversions of Asn and Gln to Asp and Glu. With the acidification of the pI, the denaturation temp. of the collagen was decreased from 42 to 35°C after 20 day treatment, but the collagen-specific triple helical conformation was maintained. Human keratinocytes and fibroblasts adhered to the alkali-treated collagen via the collagen receptor integrin α2β1. This indicates that the alkali-treated collagen maintained its property as a biol. adherent mol. Unlike acid-sol. collagen, alkali-treated collagen lost the ability to form fibrils at neutral pH under physiol. conditions. This ability was lost even after 4 h of alk. treatment, when the denaturation temp. of the collagen did not change. On the other hand, the alkali-treated collagen formed a fibrous ppt. with a uniform diam. of 50-70 nm under acidic conditions at 30°C.
- 39Yan, M.; Li, B.; Zhao, X.; Ren, G.; Zhuang, Y.; Hou, H.; Zhang, X.; Chen, L.; Fan, Y. Characterization of Acid-Soluble Collagen from the Skin of Walleye Pollock (Theragra Chalcogramma). Food Chem. 2008, 107, 1581– 1586, DOI: 10.1016/j.foodchem.2007.10.027Google ScholarThere is no corresponding record for this reference.
- 40Nurubhasha, R.; Kumar, N. S. S.; Thirumalasetti, S. K.; Simhachalam, G.; Dirisala, V. R. Extraction and Characterization of Collagen from the Skin of Pterygoplichthys Pardalis and Its Potential Application in Food Industries. Food Sci. Biotechnol. 2019, 28, 1811– 1817, DOI: 10.1007/s10068-019-00601-zGoogle Scholar40https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtVCjtbbJ&md5=3e76fd31f97de51e50bf705eafa0d3ecExtraction and characterization of collagen from the skin of Pterygoplichthys pardalis and its potential application in food industriesNurubhasha, Ramesh; Sampath Kumar, N. S.; Thirumalasetti, Satish K.; Simhachalam, G.; Dirisala, Vijaya R.Food Science and Biotechnology (2019), 28 (6), 1811-1817CODEN: FSBOBR; ISSN:1226-7708. (Korean Society of Food Science and Technology)The primary objective of this study was to ext. collagen from underutilized fish species owing to its cost effective nature and also its ability to address the demand of type I collagen arising from food and pharmaceutical industries. Acid and pepsin sol. collagen (ppASC and ppPSC) were extd. from the skin of sucker catfish (Pterygoplichthys pardalis) with a yield of 19.6 and 23.8% on wet wt. basis resp. The same were characterized and confirmed as type I collagen by SDS-PAGE, FTIR and UV-Vis spectroscopy, amino acid anal., and Zeta potential. Taking into consideration the application of collagen in food industry, a food product was developed by incorporating with fresh cheese. This fortification was found to be acceptable and had not altered the taste, odor and other sensory properties of the product.
- 41Kamalvand, M.; Biazar, E.; Daliri-Joupari, M.; Montazer, F.; Rezaei-Tavirani, M.; Heidari-Keshel, S. Design of a Decellularized Fish Skin as a Biological Scaffold for Skin Tissue Regeneration. Tissue Cell 2021, 71, 101509 DOI: 10.1016/j.tice.2021.101509Google Scholar41https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXmsFGntr0%253D&md5=b30751ec998dce878cb954d080001676Design of a decellularized fish skin as a biological scaffold for skin tissue regenerationKamalvand, Mahshad; Biazar, Esmaeil; Daliri-Joupari, Morteza; Montazer, Fatemeh; Rezaei-Tavirani, Mostafa; Heidari-Keshel, SaeedTissue & Cell (2021), 71 (), 101509CODEN: TICEBI; ISSN:0040-8166. (Elsevier Ltd.)The use of decellularized natural skin as an extracellular matrix (ECM) may be a great candidate to regenerate damaged tissues. In this study, decellularized scaffolds from fish skin were designed by different techniques (phys., chem., and enzymic methods) and investigated by analyses such as Differential Scanning Calorimetry (DSC), SEM (SEM), Tensile strength, Degradability, Histol. studies, Toxicity test, and Detn. of DNA content. Results showed that the best sample is related to the decel lularized skin by hypertonic & hypotonic technique and Triton X100 solns. Structural and mech. results were demonstrated that samples have similar properties to human skin to regenerate it. The cytotoxicity results showed that decellularized skin by hypertonic & hypotonic method and Triton soln. is non-toxic with minimal amt. of genetic materials. Cellular results with epithelial cells indicated good adhesion on decellularized matrix, so it can be a suitable candidate for skin tissue regeneration.
- 42Lu, H. D.; Soranno, D. E.; Rodell, C. B.; Kim, I. L.; Burdick, J. A. Secondary Photocrosslinking of Injectable Shear-Thinning Dock-and-Lock Hydrogels. Adv. Healthcare Mater. 2013, 2, 1028– 1036, DOI: 10.1002/adhm.201200343Google Scholar42https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhtVOkt73F&md5=9d3f88bde9acfcd51e95a83666d1419eSecondary photocrosslinking of injectable shear-thinning dock-and-lock hydrogelsLu, Hoang D.; Soranno, Danielle E.; Rodell, Christopher B.; Kim, Iris L.; Burdick, Jason A.Advanced Healthcare Materials (2013), 2 (7), 1028-1036CODEN: AHMDBJ; ISSN:2192-2640. (Wiley-VCH Verlag GmbH & Co. KGaA)Shear-thinning hydrogels are useful in numerous applications, including as injectable carriers that act as scaffolds to support cell and drug therapies. Here, we describe the engineering of a self-assembling Dock-and-Lock (DnL) system that forms injectable shear-thinning hydrogels using mol. recognition interactions that also possess photo-triggerable secondary crosslinks. These DnL hydrogels are fabricated from peptide-modified hyaluronic acid (HA) and polypeptide precursors, can self-heal immediately after shear induced flow, are cytocompatible, and can be stabilized through light-initiated radical polymn. of methacrylate functional groups to tune gel mechanics and erosion kinetics. Secondary crosslinked hydrogels retain self-adhesive properties and exhibit cooperative phys. and chem. crosslinks with moduli as high as ∼10 times larger than moduli of gels based on phys. crosslinking alone. The extent of reaction and change in properties are dependent on whether the methacrylate is incorporated either at the terminus of the peptide or directly to the HA backbone. Addnl., the gel erosion can be monitored through an incorporated fluorophore and phys.-chem. gels remain intact in soln. over months, whereas phys. gels that are not covalently crosslinked erode completely within days. Mesenchymal stem cells exhibit increased viability when cultured in phys.- chem. gels, compared with those cultured in gels based on phys. crosslinks alone. The phys. properties of these DnL gels may be addnl. tuned by adjusting component compns., which allows DnL gels with a wide range of phys. properties to be constructed for use.
- 43Guo, S.; DiPietro, L. A. Critical Review in Oral Biology & Medicine: Factors Affecting Wound Healing. J. Dent. Res. 2010, 89, 219– 229, DOI: 10.1177/0022034509359125Google Scholar43https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXjtFOnsLc%253D&md5=a42f3a6d9a5a275ced98dd32f6f7870fFactors affecting wound healingGuo, S.; Di Pietro, L. A.Journal of Dental Research (2010), 89 (3), 219-229CODEN: JDREAF; ISSN:0022-0345. (Sage Publications)A review. Wound healing, as a normal biol. process in the human body, is achieved through four precisely and highly programmed phases: hemostasis, inflammation, proliferation, and remodeling. For a wound to heal successfully, all four phases must occur in the proper sequence and time frame. Many factors can interfere with one or more phases of this process, thus causing improper or impaired wound healing. This article reviews the recent literature on the most significant factors that affect cutaneous wound healing and the potential cellular and/or mol. mechanisms involved. The factors discussed include oxygenation, infection, age and sex hormones, stress, diabetes, obesity, medications, alcoholism, smoking, and nutrition. A better understanding of the influence of these factors on repair may lead to therapeutics that improve wound healing and resolve impaired wounds.
- 44Kara, A.; Distler, T.; Polley, C.; Schneidereit, D.; Seitz, H.; Friedrich, O.; Tihminlioglu, F.; Boccaccini, A. R. 3D Printed Gelatin/Decellularized Bone Composite Scaffolds for Bone Tissue Engineering: Fabrication, Characterization and Cytocompatibility Study. Mater. Today Bio 2022, 15, 100309 DOI: 10.1016/j.mtbio.2022.100309Google Scholar44https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitFOmtbrM&md5=1ae6a6215b3ba1e06cf99b5f149c17733D printed gelatin/decellularized bone composite scaffolds for bone tissue engineering: Fabrication, characterization and cytocompatibility studyKara, Aylin; Distler, Thomas; Polley, Christian; Schneidereit, Dominik; Seitz, Hermann; Friedrich, Oliver; Tihminlioglu, Funda; Boccaccini, Aldo R.Materials Today Bio (2022), 15 (), 100309CODEN: MTBAC2; ISSN:2590-0064. (Elsevier Ltd.)Three-dimensional (3D) printing technol. enables the design of personalized scaffolds with tunable pore size and compn. Combining decellularization and 3D printing techniques provides the opportunity to fabricate scaffolds with high potential to mimic native tissue. The aim of this study is to produce novel decellularized bone extracellular matrix (dbECM)-reinforced composite-scaffold that can be used as a biomaterial for bone tissue engineering. Decellularized bone particles (dbPTs, ∼100 μm diam.) were obtained from rabbit femur and used as a reinforcement agent by mixing with gelatin (GEL) in different concns. 3D scaffolds were fabricated by using an extrusion-based bioprinter and crosslinking with microbial transglutaminase (mTG) enzyme, followed by freeze-drying to obtain porous structures. Fabricated 3D scaffolds were characterized morphol., mech., and chem. Furthermore, MC3T3-E1 mouse pre-osteoblast cells were seeded on the dbPTs reinforced GEL scaffolds (GEL/dbPTs) and cultured for 21 days to assess cytocompatibility and cell attachment. We demonstrate the 3D-printability of dbPTs-reinforced GEL hydrogels and the achievement of homogenous distribution of the dbPTs in the whole scaffold structure, as well as bioactivity and cytocompatibility of GEL/dbPTs scaffolds. It was shown that Young's modulus and degrdn. rate of scaffolds were enhanced with increasing dbPTs content. Multiphoton microscopy imaging displayed the interaction of cells with dbPTs, indicating attachment and proliferation of cells around the particles as well as into the GEL-particle hydrogels. Our results demonstrate that GEL/dbPTs hydrogel formulations have potential for bone tissue engineering.
- 45Shirazi, R.; Shirazi-Adl, A.; Hurtig, M. Role of Cartilage Collagen Fibrils Networks in Knee Joint Biomechanics under Compression. J. Biomech. 2008, 41, 3340– 3348, DOI: 10.1016/j.jbiomech.2008.09.033Google Scholar45https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BD1cjmvVGgsw%253D%253D&md5=fc8b4334d9b0ccd8f9954a0c761b51c8Role of cartilage collagen fibrils networks in knee joint biomechanics under compressionShirazi R; Shirazi-Adl A; Hurtig MJournal of biomechanics (2008), 41 (16), 3340-8 ISSN:0021-9290.Collagen fibrils networks in knee cartilage and menisci change in content and structure from a region to another. While resisting tension, they influence global joint response as well as local strains particularly at short-term periods. To investigate the role of fibrils networks in knee joint mechanics and in particular cartilage response, a novel model of the knee joint is developed that incorporates the cartilage and meniscus fibrils networks as well as depth-dependent properties in cartilage. The joint response under up to 2000N compression is investigated for conditions simulating the absence in cartilage of deep fibrils normal to subchondral bone or superficial fibrils parallel to surface as well as localized split of cartilage at subchondral junction or localized damage to superficial fibrils at loaded areas. Deep vertical fibrils network in cartilage play a crucial role in stiffening (by 10%) global response and protecting cartilage by reducing large strains (from maximum of 102% to 38%), in particular at subchondral junction. Superficial horizontal fibrils protect the tissue mainly from excessive strains at superficial layers (from 27% to 8%). Local cartilage split at base disrupts the normal function of vertical fibrils at the affected areas resulting in higher strains. Deep fibrils, and to a lesser extent superficial fibrils, play dominant mechanical roles in cartilage response under transient compression. Any treatment modality attempting to repair or regenerate cartilage defects involving partial or full thickness osteochondral grafts should account for the crucial role of collagen fibrils networks and the demanding mechanical environment of the tissue.
- 46Ying, H.; Zhou, J.; Wang, M.; Su, D.; Ma, Q.; Lv, G.; Chen, J. In Situ Formed Collagen-Hyaluronic Acid Hydrogel as Biomimetic Dressing for Promoting Spontaneous Wound Healing. Mater. Sci. Eng. C 2019, 101, 487– 498, DOI: 10.1016/j.msec.2019.03.093Google Scholar46https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXntFens7c%253D&md5=5c646f9422649322d527fd0d953f4c61In situ formed collagen-hyaluronic acid hydrogel as biomimetic dressing for promoting spontaneous wound healingYing, Huiyan; Zhou, Juan; Wang, Mingyu; Su, Dandan; Ma, Qiaoqiao; Lv, Guozhong; Chen, JinghuaMaterials Science & Engineering, C: Materials for Biological Applications (2019), 101 (), 487-498CODEN: MSCEEE; ISSN:0928-4931. (Elsevier B.V.)Wound dressing is distinctly important to wound healing, because it can not only protect wound from external disturbance, but also provide an ideal environment for wound closure. However, most of wound dressings need addnl. active ingredients to assist the repair process. In order to develop new dressings that can present spontaneous healing activity, herein, an injectable hydrogel consisted of collagen I and hyaluronic acid has been designed to mimic extracellular matrix for vascular cells growing and wound closure. The prepn. of hydrogel (COL-HA) was realized through in situ coupling of phenol moieties of collagen I-hydroxybenzoic acid (COL-P) and hyaluronic-acid-tyramine (HA-Tyr) through horseradish peroxidase (HRP). The phys. structure and properties were characterized, and the biol. performances were analyzed. COL-HA hydrogel presented porous structure that contributed to the exchange of gas, medium and nutrition. Human microvascular endothelial cells (HMEC) and fibroblasts (COS-7) cultured within this hydrogel showed significant proliferation behaviors. More importantly, a certain level of vascular endothelial growth factor (VEGF) was obsd. in HMEC cultured hydrogel, which led to the possibility of vascular regeneration. For the full-thickness wound, the healing ratio and validity of wound treated with COL-HA hydrogel were higher than com. drug and individual COL-P hydrogel, HA-Tyr hydrogel groups, since collagen and hyaluronic acid made joint efforts to improve wound repair.
- 47Uhl, F. E.; Zhang, F.; Pouliot, R. A.; Uriarte, J. J.; Rolandsson Enes, S.; Han, X.; Ouyang, Y.; Xia, K.; Westergren-Thorsson, G.; Malmström, A.; Hallgren, O.; Linhardt, R. J.; Weiss, D. J. Functional Role of Glycosaminoglycans in Decellularized Lung Extracellular Matrix. Acta Biomater. 2020, 102, 231– 246, DOI: 10.1016/j.actbio.2019.11.029Google Scholar47https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXitlehtL%252FO&md5=9b4dff340393b04d139016fd37ddaba6Functional role of glycosaminoglycans in decellularized lung extracellular matrixUhl, Franziska E.; Zhang, Fuming; Pouliot, Robert A.; Uriarte, Juan J.; Rolandsson Enes, Sara; Han, Xiaorui; Ouyang, Yilan; Xia, Ke; Westergren-Thorsson, Gunilla; Malmstrom, Anders; Hallgren, Oskar; Linhardt, Robert J.; Weiss, Daniel J.Acta Biomaterialia (2020), 102 (), 231-246CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Despite progress in use of decellularized lung scaffolds in ex vivo lung bioengineering schemes, including use of gels and other materials derived from the scaffolds, the detailed compn. and functional role of extracellular matrix (ECM) proteoglycans (PGs) and their glycosaminoglycan (GAG) chains remaining in decellularized lungs, is poorly understood. Using a commonly utilized detergent-based decellularization approach in human autopsy lungs resulted in disproportionate losses of GAGs with depletion of chondroitin sulfate/dermatan sulfate (CS/DS) > heparan sulfate (HS) > hyaluronic acid (HA). Specific changes in disaccharide compn. of remaining GAGs were obsd. with disproportionate loss of NS and NS2S for HS groups and of 4S for CS/DS groups. No significant influence of smoking history, sex, time to autopsy, or age was obsd. in native vs. decellularized lungs. Notably, surface plasmon resonance demonstrated that GAGs remaining in decellularized lungs were unable to bind key matrix-assocd. growth factors FGF2, HGF, and TGFβ1. Growth of lung epithelial, pulmonary vascular, and stromal cells cultured on the surface of or embedded within gels derived from decellularized human lungs was differentially and combinatorially enhanced by replenishing specific GAGs and FGF2, HGF, and TGFβ1. In summary, lung decellularization results in loss and/or dysfunction of specific GAGs or side chains significantly affecting matrix-assocd. growth factor binding and lung cell metab. GAG and matrix-assocd. growth factor replenishment thus needs to be incorporated into schemes for investigations utilizing gels and other materials produced from decellularized human lungs. In the current studies, we demonstrate that glycosaminoglycans (GAGs) are significantly depleted during decellularization and those that remain are dysfunctional and unable to bind matrix-assocd. growth factors crit. for cell growth and differentiation. Systematically repleting GAGs and matrix-assocd. growth factors to gels derived from decellularized human lung significantly and differentially affects cell growth. These studies highlight the importance of considering GAGs in decellularized lungs and their derivs.
- 48Linhardt, R. J.; Toida, T. Linhardt.ReviewGAG.121130.Pdf. Acc. Chem. Res. 2004, 37, 431– 438, DOI: 10.1021/ar030138xGoogle Scholar48https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXjvVShtbk%253D&md5=2211c39d916084933178e816466cedeeRole of glycosaminoglycans in cellular communicationLinhardt, Robert J.; Toida, ToshihikoAccounts of Chemical Research (2004), 37 (7), 431-438CODEN: ACHRE4; ISSN:0001-4842. (American Chemical Society)A review. Glycosaminoglycans are of crit. importance in intercellular communication in organisms. This ubiquitous class of linear polyanions interacts with a wide variety of proteins, including growth factors and chemokines, which regulate important physiol. processes. The presence of glycosaminoglycans on cell membranes and in the extracellular matrix also has resulted in their exploitation by infectious pathogens to gain access and entry into animal cells. Here, the authors have examd. the structural and phys. characteristics of these mols. responsible for their interaction with proteins important in cell-cell communication.
- 49West, J. D.; West, J. D.; Stamm, C. E.; Brown, H. A.; Justice, S. L.; Morano, K. A. Enhanced Toxicity of the Protein Cross-Linkers Divinyl Sulfone and Diethyl Acetylenedicarboxylate in Comparison to Related Monofunctional Electrophiles. Chem. Res. Toxicol. 2011, 24, 1457– 1459, DOI: 10.1021/tx200302wGoogle Scholar49https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXpvVygs74%253D&md5=b87c2368fb55d2f4d7de6f0c96f853fbEnhanced Toxicity of the Protein Cross-Linkers Divinyl Sulfone and Diethyl Acetylenedicarboxylate in Comparison to Related Monofunctional ElectrophilesWest, James D.; Stamm, Chelsea E.; Brown, Haley A.; Justice, Samantha L.; Morano, Kevin A.Chemical Research in Toxicology (2011), 24 (9), 1457-1459CODEN: CRTOEC; ISSN:0893-228X. (American Chemical Society)Previously, the authors detd. that di-Et acetylenedicarboxylate (DAD), a protein cross-linker, was significantly more toxic than analogous monofunctional electrophiles. The authors hypothesized that other protein cross-linkers enhance toxicity similarly. In agreement with this hypothesis, the bifunctional electrophile divinyl sulfone (DVSF) was 6-fold more toxic than Et vinyl sulfone (EVSF) in colorectal carcinoma cells and greater than 10-fold more toxic in Saccharomyces cerevisiae. DVSF and DAD caused oligomerization of yeast thioredoxin 2 (Trx2p) in vitro and promoted Trx2p crosslinking to other proteins in yeast at cytotoxic doses. These results suggest that protein crosslinking is considerably more detrimental to cellular homeostasis than simple alkylation.
- 50Fang, Y.; Han, Y.; Wang, S.; Chen, J.; Dai, K.; Xiong, Y.; Sun, B. Three-Dimensional Printing Bilayer Membranous Nanofiber Scaffold for Inhibiting Scar Hyperplasia of Skin. Biomater. Adv. 2022, 138, 212951 DOI: 10.1016/j.bioadv.2022.212951Google Scholar50https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitlOmu73P&md5=7bd7f1f553d62782a322eb27c6e56bc5Three-dimensional printing bilayer membranous nanofiber scaffold for inhibiting scar hyperplasia of skinFang, Yuan; Han, Yu; Wang, Shoubao; Chen, Jingting; Dai, Kerong; Xiong, Yao; Sun, BinbinBiomaterials Advances (2022), 138 (), 212951CODEN: BAIDBT; ISSN:2772-9508. (Elsevier B.V.)Hypertrophic scar (HS) is the manifestation of pathol. wound healing, which affects the beauty of patients, and even affects the normal phys. functions of patients. We aimed to develop a 3D printing layer membranous nanofiber scaffold similar to skin structure. Among them, poly (lactic-co-glycolic acid) copolymer (PLGA) nanofibers were used as the "epidermis" layer above, and a decellular dermis matrix (dECM) nanofiber scaffold was used as the "dermis" layer below. In vitro, exptl. results showed that PLGA and dECM nanofiber scaffolds had good biocompatibility. In vivo expts. showed that BLM nanofiber scaffolds could inhibit collagen fiber deposition and angiogenesis, to inhibit the formation of hypertrophic scars. This study shows a simple and effective method for preventing and inhibiting the formation of hypertrophic scars.
- 51Zhang, T.; Xu, H.; Zhang, Y.; Zhang, S.; Yang, X.; Wei, Y.; Huang, D.; Lian, X. Fabrication and Characterization of Double-Layer Asymmetric Dressing through Electrostatic Spinning and 3D Printing for Skin Wound Repair. Mater. Des. 2022, 218, 110711 DOI: 10.1016/j.matdes.2022.110711Google Scholar51https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38Xhtlams7rK&md5=8cdc5b7b68a3235097081a0f1a0b6395Fabrication and characterization of double-layer asymmetric dressing through electrostatic spinning and 3D printing for skin wound repairZhang, Ting; Xu, Hao; Zhang, Yonggang; Zhang, Siruo; Yang, Xia; Wei, Yan; Huang, Di; Lian, XiaojieMaterials & Design (2022), 218 (), 110711CODEN: MADSD2; ISSN:0264-1275. (Elsevier Ltd.)Ideal wound dressings provide optimal microenvironment for the reconstruction of damaged tissue. In this work, we fabricated a bilayer asym. dressing to mimic gradient structure of epidermis and dermis of skin by combining electrostatic spinning and 3D printing method with properties including surface hydrophilic and hydrophobic, porosity, mech. as well as antibacterial properties. The outer layer was prepd. by optimized PCL/PLA (PP) via electrostatic spinning to mimick epidermis with water repellency and against bacterial penetration, which has a tensile modulus of 19.69 ± 0.66 MPa. While, the inner layer was 3D printed by optimized sodium alginate/polyvinyl alc./chitosan quaternary ammonium salt (SPH). The tensile modulus of SPH with a porosity of 70-90% is 0.82 ± 0.01 MPa, and the water content can be achieved above 85%. The antibacterial efficacy of inner layer was tested against Staphylococcus aureus indicating forming inhibition zone with a diam. of 1.61 ± 0.35 cm. In addn., Cell Counting Kit-8 and Live/Dead assay was used to test the viability of human dermal fibroblasts (HFBS), which showed that PP/SPH with 6% PVA had not significant cytotoxic effects. The double-layer asym. dressing meets the requirements of skin mech. properties and provides an effective repair strategy for clin. skin trauma.
- 52Randall, M. J.; Jüngel, A.; Rimann, M.; Wuertz-Kozak, K. Advances in the Biofabrication of 3D Skin in Vitro: Healthy and Pathological Models. Front. Bioeng. Biotechnol. 2018, 6, 154 DOI: 10.3389/fbioe.2018.00154Google Scholar52https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3crgtFOksg%253D%253D&md5=63c5c7c795fc0b45203ba69b660e754cAdvances in the Biofabrication of 3D Skin in vitro: Healthy and Pathological ModelsRandall Matthew J; Wuertz-Kozak Karin; Jungel Astrid; Rimann Markus; Rimann Markus; Wuertz-Kozak Karin; Wuertz-Kozak KarinFrontiers in bioengineering and biotechnology (2018), 6 (), 154 ISSN:2296-4185.The relevance for in vitro three-dimensional (3D) tissue culture of skin has been present for almost a century. From using skin biopsies in organ culture, to vascularized organotypic full-thickness reconstructed human skin equivalents, in vitro tissue regeneration of 3D skin has reached a golden era. However, the reconstruction of 3D skin still has room to grow and develop. The need for reproducible methodology, physiological structures and tissue architecture, and perfusable vasculature are only recently becoming a reality, though the addition of more complex structures such as glands and tactile corpuscles require advanced technologies. In this review, we will discuss the current methodology for biofabrication of 3D skin models and highlight the advantages and disadvantages of the existing systems as well as emphasize how new techniques can aid in the production of a truly physiologically relevant skin construct for preclinical innovation.
- 53Kim, S.; Lee, H.; Kim, J. A.; Park, T. H. Prevention of Collagen Hydrogel Contraction Using Polydopamine-Coating and Alginate Outer Shell Increases Cell Contractile Force. Biomater. Adv. 2022, 136, 212780 DOI: 10.1016/j.bioadv.2022.212780Google Scholar53https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitlOmurvP&md5=2d788e46a6d51bde1e6116e1f8b80d85Prevention of collagen hydrogel contraction using polydopamine-coating and alginate outer shell increases cell contractile forceKim, Seulha; Lee, Haein; Kim, Jeong Ah.; Park, Tai HyunBiomaterials Advances (2022), 136 (), 212780CODEN: BAIDBT; ISSN:2772-9508. (Elsevier B.V.)Collagen is the most abundant protein in the extracellular matrix of mammals and has a great effect on various cell behaviors including adhesion, differentiation, and migration. However, it is difficult to utilize collagen gel as a phys. scaffold in vitro because of its severe contraction. Decrease in the overall hydrogel vol. induces changes in cell distribution, and mass transfer within the gel. Uncontrolled mech. and physiol. factors in the fibrous matrix result in uncontrolled cell behaviors in the surrounding cells. In this study, two strategies were used to minimize the contraction of collagen gel. A disk-shaped frame made of polydopamine-coated polydimethylsiloxane (PDMS) prevented horizontal contraction at the edge of the hydrogel. The sequentially cross-linked collagen gel with alginate outer shell (CA-shell) structure inhibited the vertical gel contraction. The combined method synergistically prevented the hydrogel from shrinkage in long-term 3D cell culture. We obsd. the shift in balance of differentiation from adipogenesis to osteogenesis in mesenchymal stem cells under the environment where gel contraction was prevented, and confirmed that this phenomenon is closely assocd. with the mechanotransduction based on Yes-assocd. protein (YAP) localization. Development of this contraction inhibition platform made it possible to investigate the influence of regulation of cellular microenvironments. The phys. properties of the hydrogel fabricated in this study were similar to that of pure collagen gel but completely changed the cell behavior within the gel by inhibition of gel contraction. The platform can be used to broaden our understanding of the fundamental mechanism underlying cell-matrix interactions and reproduce extracellular matrix in vivo.
- 54Bacakova, M.; Pajorova, J.; Broz, A.; Hadraba, D.; Lopot, F.; Zavadakova, A.; Vistejnova, L.; Kostic, I.; Jencova, V.; Bacakova, L. Erratum to a Two-Layer Skin Construct Consisting of a Collagen Hydrogel Reinforced by a Fibrin-Coated Polylactide Nanofibrous Membrane (Int J Nanomedicine, (2019), 14, (5033–5050)). Int. J. Nanomed. 2019, 14, 7215– 7216, DOI: 10.2147/IJN.S228110Google ScholarThere is no corresponding record for this reference.
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This article is cited by 6 publications.
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Abstract
Figure 1
Figure 1. Schematic diagram of the preparation of solubilized fish-dECM and 3D hybrid bioink for cocultivation of HDF and HaCaT cells with a 3D-printed model in an air–liquid condition. (a) Solubilization process of fish-dECM in alkaline conditions. (b) Preparation of 3D hybrid bioink (pre-gel) and fabrication of a 3D-printed model via 3D printing and secondary crosslinking of the 3D scaffold.
Figure 2
Figure 2. Characterization of dECM. (a) UV–vis spectra of solubilized fish-dECM before or after dialysis. Absorbance was normalized to total protein determined by a BCA assay. (b) Cytotoxicity of fish-dECM eluant. Fish-dECM (6 cm2) was eluated with DMEM supplemented with 10% of FBS (1 mL), followed by mixing with cell cultivation medium at different mixing ratios (0–100%) and addition to NIH3T3 cells (1 × 104 cells/well (surface area = 0.32 cm2)). After 24 h, an MTT-based cytotoxicity assay was performed and the viability of the treated cells was normalized with respect to those without treatment.
Figure 3
Figure 3. Investigation of injectability and printability of different HA-based bioinks, and characterization of 3D-printed models. (a, b) Optimization of 3D hybrid bioink. Injectability and printability of 3D hybrid bioink crosslinked at various DVS molar ratios (molar ratio of HA:DVS = 1:0.020–0.055) were examined (O, high; Δ, medium; X, low injectability/printability). (c) Images of the custom design model and 3D-printed model (scale bar = 1 cm). (d) Water swelling ratio of the 3D-printed model with 7% (v/v) DVS (n = 3). (e) Mass erosion of 3D-printed model; 3D-printed model was immersed in PBS (pH 7.4) at 37 °C for 7, 14, and 21 d. At the designated time point, the degraded 3D model was washed thrice with DW and freeze-dried; the dried 3D model weight was recorded (n = 3). *indicates a significant difference between HA/0dECM, HA/10dECM, and HA/30dECM; p < 0.05 were considered statistically significant.
Figure 4
Figure 4. In vitro studies on cell attachment and cell viability of HA-based 3D-printed scaffolds. (a) NIH3T3 cells attachment on 3D-printed models. Unbound cells were stained with trypan blue and counted using a hemocytometer. Cell attachment* (%) = ((number of initial cells – counted cells)/number of initial cells × 100). (b) Proliferation of cells cultivated on 3D-printed models for 3, 5, and 7 d (n = 3). Cell proliferation was determined by a WST-1-based colorimetric assay. * indicates a significant difference between HA/0dECM, HA/10dECM, and HA/30dECM; p < 0.05 were considered statistically significant.
Figure 5
Figure 5. HDF and HaCaT cocultivation on 3D-printed scaffolds. (a) Cocultivation of the HDF/HaCaT/3D-printed model. After 5 d of incubation with HDF cells, HaCaT cells were added and incubated for another 5 days in air–liquid conditions to induce keratinization of HaCaT cells. (b) Masson’s trichrome and hematoxylin staining of HDF and HaCaT-cocultivated 3D scaffolds. Collagens (white arrows) and cell nuclei (white circles) are stained in blue and violet. Dashed lines separate the regions of the epidermis (ED) and the dermis (D) (scale bar = 100 μm).
References
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- 1Kiyotake, E. A.; Douglas, A. W.; Thomas, E. E.; Nimmo, S. L.; Detamore, M. S. Development and Quantitative Characterization of the Precursor Rheology of Hyaluronic Acid Hydrogels for Bioprinting. Acta Biomater. 2019, 95, 176– 187, DOI: 10.1016/j.actbio.2019.01.0411https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvFeksrY%253D&md5=fdae8ca8387f8c70ad58c7ddfd713dcbDevelopment and quantitative characterization of the precursor rheology of hyaluronic acid hydrogels for bioprintingKiyotake, Emi A.; Douglas, Alexander W.; Thomas, Emily E.; Detamore, Michael S.Acta Biomaterialia (2019), 95 (), 176-187CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Bioprinting technologies have tremendous potential for advancing regenerative medicine due to the precise spatial control over depositing a printable biomaterial, or bioink. Despite the growing interest in bioprinting, the field is challenged with developing biomaterials for extrusion-based bioprinting. The paradigm of contemporary bioink studies relies on trial-and-error methods for discovering printable biomaterials, which has little practical use for others who endeavor to develop bioinks. There is pressing need to follow the precedent set by a few pioneering studies that have attempted to standardize bioink characterizations for detg. the properties that define printability. Here, we developed a pentenoate-functionalized hyaluronic acid hydrogel (PHA) into a printable bioink and used three recommended, quant. rheol. assessments to characterize the printability: (1) yield stress, (2) viscosity, and (3) storage modulus recovery. The most important characteristic is the yield stress; we found a yield stress upper limit of ∼1000 Pa for PHA. Measuring the viscosity was advantageous for detg. shear-thinning behavior, which aided in extruding highly viscous PHA through a nozzle. Post-printing recovery is required to maintain shape fidelity and we found storage modulus recoveries above ∼85% were sufficient for PHA. Two formulations had superior printability (i.e., 1.5 MDa PHA - 4 wt%, and 1 MDa PHA - 8 wt%), and increasing cell concns. in PHA up to 9 × 106 cells/mL had minimal effects on the printability. Even so, other factors such as sterilization and peptide modifications to enhance bioactivity may influence printability, highlighting the need for investigators to consider such factors when developing new bioinks. Bioprinting has potential for regenerating damaged tissues; however, there are a limited no. of printable biomaterials, and developing new bioinks is challenging because the required material phys. properties for extrusion-based printing are not yet known. Most new bioinks are developed by trial-and-error, which is neither efficient nor comparable across materials. There is a need for the field to begin utilizing std. methods proposed by a few pioneering studies to characterize new bioinks. Therefore, we have developed the printability of a hyaluronic acid based-hydrogel and characterized the material with three quant. rheol. tests. The current work impacts the bioprinting field by demonstrating and encouraging the use of universal bioink characterizations and by providing printability windows to advance new bioink development.
- 2Ge, B.; Wang, H.; Li, J.; Liu, H.; Yin, Y.; Zhang, N.; Qin, S. Comprehensive Assessment of Nile Tilapia Skin. Mar. Drugs 2021, 18, 178– 194, DOI: 10.3390/md18040178There is no corresponding record for this reference.
- 3Ouyang, L.; Highley, C. B.; Rodell, C. B.; Sun, W.; Burdick, J. A. 3D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-Linking. ACS Biomater. Sci. Eng. 2016, 2, 1743– 1751, DOI: 10.1021/acsbiomaterials.6b001583https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xos1Skt7c%253D&md5=870810033017a9e057780d8e4f861e963D Printing of Shear-Thinning Hyaluronic Acid Hydrogels with Secondary Cross-LinkingOuyang, Liliang; Highley, Christopher B.; Rodell, Christopher B.; Sun, Wei; Burdick, Jason A.ACS Biomaterials Science & Engineering (2016), 2 (10), 1743-1751CODEN: ABSEBA; ISSN:2373-9878. (American Chemical Society)The development of printable biomaterial inks is crit. to the application of 3D printing in biomedicine. To print high-resoln. structures with fidelity to a computer-aided design, materials used in 3D printing must be capable of being deposited on a surface and maintaining a printed structure. A dual-crosslinking hyaluronic acid system was studied here as a printable hydrogel ink, which encompassed both shear-thinning and self-healing behaviors via guest-host bonding, as well as covalent crosslinking for stabilization using photopolymn. When either guest-host assembly or covalent crosslinking was used alone, long-term stable structures were not formed, because of network relaxation after printing or dispersion of the ink filaments prior to stabilization, resp. The dual-crosslinking hydrogel filaments formed structures with greater than 16 layers that were stable over a month with no loss in mech. properties and the printed filament size ranged from 100 to 500 μm, depending on printing parameters (needle size, speed, and extrusion flux). Printed structures were further functionalized (i.e., RGD peptide) to support cell adhesion. This work highlights the importance of ink formulation and crosslinking on the printing of stable hydrogel structures.
- 4Petta, D.; Armiento, A. R.; Grijpma, D.; Alini, M.; Eglin, D.; D’Este, M. 3D Bioprinting of a Hyaluronan Bioink through Enzymatic-and Visible Light-Crosslinking. Biofabrication 2018, 10, 044104 DOI: 10.1088/1758-5090/aadf584https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXmtFSitr0%253D&md5=c99806172bc4676bb93f6b30d6b3ed6cThree dimensional bioprinting of a hyaluronan bioink through enzymatic-and visible light-crosslinkingPetta, D.; Armiento, A. R.; Grijpma, D.; Alini, M.; Eglin, D.; D'Este, M.Biofabrication (2018), 10 (4), 044104/1-044104/11CODEN: BIOFFN; ISSN:1758-5090. (IOP Publishing Ltd.)Extrusion-based three-dimensional bioprinting relies on bioinks engineered to combine viscoelastic properties for extrusion and shape retention, and biol. properties for cytocompatibility and tissue regeneration. To satisfy these conflicting requirements, bioinks often utilize either complex mixts. or complex modifications of biopolymers. In this paper we introduce and characterize a bioink exploiting a dual crosslinking mechanism, where an enzymic reaction forms a soft gel suitable for cell encapsulation and extrusion, while a visible light photo-crosslinking allows shape retention of the printed construct. The influence of cell d. and cell type on the rheol. and printability properties was assessed correlating the printing outcomes with the damping factor, a rheol. characteristic independent of the printing system. Stem cells, chondrocytes and fibroblasts were encapsulated, and their viability was assessed up to 14 days with live/dead, alamar blue and trypan blue assays. Addnl., the impact of the printing parameters on cell viability was investigated. Owing to its straightforward prepn., low modification, presence of two independent crosslinking mechanisms for tuning shear-thinning independently of the final shape fixation, the use of visible green instead of UV light, the possibility of encapsulating and sustaining the viability of different cell types, the hyaluronan bioink here presented is a valid biofabrication tool for producing 3D printed tissue-engineered constructs.
- 5Petta, D.; D’Amora, U.; Ambrosio, L.; Grijpma, D. W.; Eglin, D.; D’Este, M. Hyaluronic Acid as a Bioink for Extrusion-Based 3D Printing. Biofabrication 2020, 12, 032001 DOI: 10.1088/1758-5090/ab87525https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXitFCisr%252FN&md5=afa69e4f9418987ed3741ae1665a74aeHyaluronic acid as a bioink for extrusion-based 3D printingPetta, D.; D'amora, U.; Ambrosio, L.; Grijpma, D. W.; Eglin, D.; D'este, M.Biofabrication (2020), 12 (3), 032001CODEN: BIOFFN; ISSN:1758-5090. (IOP Publishing Ltd.)A review. Biofabrication is enriching the tissue engineering field with new ways of producing structurally organized complex tissues. Among the numerous bioinks under investigation, hyaluronic acid (HA) and its derivs. stand out for their biol. relevance, cytocompatibility, shear-thinning properties, and potential to fine-tune the desired properties with chem. modification. In this paper, we review the recent advances on bioinks contg. HA. The available literature is presented based on subjects including the rheol. properties in connection with printability, the chem. strategies for endowing HA with the desired properties, the clin. application, the most advanced preclin. studies, the advantages and limitations in comparison with similar biopolymer-based bioinks, and future perspectives.
- 6Qiao, Y.; Xu, S.; Zhu, T.; Tang, N.; Bai, X.; Zheng, C. Preparation of Printable Double-Network Hydrogels with Rapid Self-Healing and High Elasticity Based on Hyaluronic Acid for Controlled Drug Release. Polymer 2020, 186, 121994 DOI: 10.1016/j.polymer.2019.1219946https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXit1CmurzL&md5=183ff4e2d9197f1926fd150df38e3b1aPreparation of printable double-network hydrogels with rapid self-healing and high elasticity based on hyaluronic acid for controlled drug releaseQiao, Yang; Xu, Shichao; Zhu, Tianzhe; Tang, Nan; Bai, Xuejian; Zheng, ChunmingPolymer (2020), 186 (), 121994CODEN: POLMAG; ISSN:0032-3861. (Elsevier Ltd.)On account of the features about computer-accurate control, the technol. of 3D printing has a broad future for developing customized ergonomic biomedical materials. Herein, a printable dual-network hydrogel for drug delivery was designed. The acrylamide-modified hyaluronic acid was mixed with folic acid and Fe3+ at first. With the formation of metal-carboxylate coordination bonds, the first layer of phys. crosslinking network (HFF) was formed, followed by performing the spectral anal. and rheol. measurements to verify the bonding. The second layer of PAM network was then polymd. by UV radiation, with high elasticity and fatigue resistance to satisfy its application as a wound dressing. Furthermore, acetylsalicylic acid (Asa) was used as a drug model resulted in a pH responsiveness of the prepd. hydrogels with the sustained drug releasing behavior over 300 h. These results signified the potential application of this self-healing hydrogel with good mech. properties in regenerative medicine.
- 7Antich, C.; de Vicente, J.; Jiménez, G.; Chocarro, C.; Carrillo, E.; Montañez, E.; Gálvez-Martín, P.; Marchal, J. A. Bio-Inspired Hydrogel Composed of Hyaluronic Acid and Alginate as a Potential Bioink for 3D Bioprinting of Articular Cartilage Engineering Constructs. Acta Biomater. 2020, 106, 114– 123, DOI: 10.1016/j.actbio.2020.01.0467https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjtVanu74%253D&md5=8df42b62707737a82d3a9687849b11c4Bio-inspired hydrogel composed of hyaluronic acid and alginate as a potential bioink for 3D bioprinting of articular cartilage engineering constructsAntich, Cristina; de Vicente, Juan; Jimenez, Gema; Chocarro, Carlos; Carrillo, Esmeralda; Montanez, Elvira; Galvez-Martin, Patricia; Marchal, Juan AntonioActa Biomaterialia (2020), 106 (), 114-123CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Bioprinting is a promising tool to fabricate well-organized cell-laden constructs for repair and regeneration of articular cartilage. In this study, we focused on the use of one of the main cartilage components, hyaluronic acid (HA), to design and formulate a new bioink for cartilage tissue 3D bioprinting. To produce cartilage constructs with optimal mech. properties, HA-based bioink was co-printed with polylactic acid (PLA). HA-based bioink was found to improve cell functionality by an increase in the expression of chondrogenic gene markers and specific matrix deposition and, therefore, tissue formation. It is a promising bioink candidate for cartilage tissue engineering based in 3D bioprinting. The recent appearance of 3D printing technol. has enabled great advances in the treatment of osteochondral disorders by fabrication of cartilage tissue constructs that restore and/or regenerate damaged tissue. In this study, we describe for first time the development of a bioink based on the main component of cartilage, HA, with suitable biol. and mech. properties, without involving toxic procedure, and its application in cartilage tissue bioprinting. Hybrid constructs prepd. by co-printing this bioink and thermoplastic polymer PLA provided an optimal niche for chondrocyte growth and maintenance as well as mech. properties necessary to support load forces exerted in native tissue. We highlight the translation potential of this HA-based bioink in the clin. arena.
- 8Roushangar Zineh, B.; Shabgard, M. R.; Roshangar, L.; Jahani, K. Experimental and Numerical Study on the Performance of Printed Alginate/Hyaluronic Acid/Halloysite Nanotube/Polyvinylidene Fluoride Bio-Scaffolds. J. Biomech. 2020, 104, 109764 DOI: 10.1016/j.jbiomech.2020.1097648https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB38zgvFGksQ%253D%253D&md5=b5e5f522b57a0b2a4029939280f869b7Experimental and numerical study on the performance of printed alginate/hyaluronic acid/halloysite nanotube/polyvinylidene fluoride bio-scaffoldsRoushangar Zineh Babak; Jahani Kamal; Shabgard Mohammad Reza; Roshangar LeilaJournal of biomechanics (2020), 104 (), 109764 ISSN:.The growing usage of printed bio scaffolds in the field of regenerative medicine has made this field very important in biomedical engineering. In this regard, three-dimensional printing (3D) technique needs bio-materials with higher mechanical and biological performance. The biomaterials with high mechanical performance beside its bio compatibility are limited. A novel bio-material made of Alginate, Hyaluronic acid, Halloysite Nanotube and Polyvinylidene Fluoride was used and characterized for printing cartilage bio scaffolds through numerical studies. CaCl2 was used for crosslinking of biomaterial. Scanning Electron Microscopy, mechanical tests (tensile and compressive test), MTT assay were conducted for evaluating this novel biomaterial. Different structures of bio material were simulated for numerical studies. The numerical study was performed in ANSYS 18 using three parameter Mooney-Rivlin model. According to experimental and numerical results, Halloysite Nanotube increases the tensile and compressive strength of biomaterial up to 47%. Results show that biomaterial have good mechanical performance due to mechanical forces required for cartilage bio scaffolds besides its high biological performance. Polyvinylidene fluoride reduces the mechanical performance while increasing the cell viability. MTT assay results performed on day 0, day 2 and day 6 show increase in cell number to be about twice for biomaterial containing 40 mg/ml alginate, 40 mg/ml halloysite nanotube, 10 mg/ml hyaluronic acid and 1 w/v Polyvinylidene fluoride. Numerical simulation shows high mechanical performance of bio material in different scaffolds structure. The best structure of bio scaffolds was achieved with 0.4 mm nozzle diameter and 0.4 space between rows.
- 9Pérez, L. A.; Hernández, R.; Alonso, J. M.; Pérez-González, R.; Sáez-Martínez, V. Hyaluronic Acid Hydrogels Crosslinked in Physiological Conditions: Synthesis and Biomedical Applications. Biomedicines 2021, 9, 1113 DOI: 10.3390/biomedicines90911139https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXisVyqtb3I&md5=8bc04f98b8ce3998dccf9808ca1149f7Hyaluronic Acid Hydrogels Crosslinked in Physiological Conditions: Synthesis and Biomedical ApplicationsPerez, Luis Andres; Hernandez, Rebeca; Alonso, Jose Maria; Perez-Gonzalez, Raul; Saez-Martinez, VirginiaBiomedicines (2021), 9 (9), 1113CODEN: BIOMID; ISSN:2227-9059. (MDPI AG)Hyaluronic acid (HA) hydrogels display a wide variety of biomedical applications ranging from tissue engineering to drug vehiculization and controlled release. To date, most of the com. available hyaluronic acid hydrogel formulations are produced under conditions that are not compatible with physiol. ones. This review compiles the currently used approaches for the development of hyaluronic acid hydrogels under physiol./mild conditions. These methods include dynamic covalent processes such as boronic ester and Schiff-base formation and click chem. mediated reactions such as thiol chem. processes, azide-alkyne, or Diels Alder cycloaddn. Thermoreversible gelation of HA hydrogels at physiol. temp. is also discussed. Finally, the most outstanding biomedical applications are indicated for each of the HA hydrogel generation approaches.
- 10Yang, R.; Tan, L.; Cen, L.; Zhang, Z. An Injectable Scaffold Based on Crosslinked Hyaluronic Acid Gel for Tissue Regeneration. RSC Adv. 2016, 6, 16838– 16850, DOI: 10.1039/c5ra27870h10https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xit1Orsb4%253D&md5=c915a516700130b50ceeae0721fa2660An injectable scaffold based on crosslinked hyaluronic acid gel for tissue regenerationYang, Rui; Tan, Linhua; Cen, Lian; Zhang, ZhibingRSC Advances (2016), 6 (20), 16838-16850CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)Injectable scaffolds have great potential in specialised applications in regenerative medicine. In this study, hyaluronic acid hydrogels (HAGs) were prepd. by crosslinking hyaluronic acid (HA) with 1,4-butanediol diglycidyl ether (BDDE). Applications of HAG as an injectable scaffold for regenerating functional tissues were proposed by matching its viscoelastic properties with those of biol. tissues. The effect of BDDE concn. on different properties of HAGs was explored. Swelling properties, cross-sectional morphol., and BDDE residues of the resulting gels were investigated. Rheol. properties of different HAGs were measured by monitoring their storage modulus (G') and loss modulus (G'') and compared with those of biol. tissues. It was shown that HAGs (BDDE from 0.4 vol% to 1.0 vol%) possess great water absorbing capability with swelling ratios ranging from 99.7 to 78.9. The higher the concn. of the crosslinker used, the more rigid the resulting hydrogel, subsequently the lower the swelling ratio would be and the higher the G' and G'' values as well. Similar viscoelastic behaviors were found between HAGs and biol. tissues, such as epidermis, dermis, articular cartilage and tooth germ. SEM revealed that HAG obtained at 0.4 vol% BDDE had pore diams. ranging from a few microns to around 100 μm with a high degree of interconnectivity. The feasibility of this HAG, as an injectable scaffold, to regenerate cartilage and dentin-pulp complex was then demonstrated using a preliminary s.c. microenvironment. The current study could be a ref. to take account of how a crosslinked HA gel should be chosen for specific tissue regeneration.
- 11Collins, M. N.; Birkinshaw, C. Hyaluronic Acid Based Scaffolds for Tissue Engineering - A Review. Carbohydr. Polym. 2013, 92, 1262– 1279, DOI: 10.1016/j.carbpol.2012.10.02811https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXisVSms7c%253D&md5=842c3465f2505666f3c814430e7539ffHyaluronic acid based scaffolds for tissue engineering-A reviewCollins, Maurice N.; Birkinshaw, ColinCarbohydrate Polymers (2013), 92 (2), 1262-1279CODEN: CAPOD8; ISSN:0144-8617. (Elsevier Ltd.)This review focuses on hyaluronic acid (HA) tissue scaffolding materials. Scaffolds are defined in terms of formation mechanisms and mode of action. Soln. properties are discussed as an understanding of the hydrodynamics of HA is fundamental in optimizing the subsequent modification and the chemistries behind important tissue engineering applications that are emerging from recent research on this increasingly valuable carbohydrate polymer are described. Key scaffold characteristics such as mech., biol. function and degrdn. are discussed. The latest technologies behind scaffold processing are assessed and the applications of HA based scaffolds are discussed.
- 12Khunmanee, S.; Jeong, Y.; Park, H. Crosslinking Method of Hyaluronic-Based Hydrogel for Biomedical Applications. J. Tissue Eng. 2017, 8, 204173141772646 DOI: 10.1177/2041731417726464There is no corresponding record for this reference.
- 13Seif-Naraghi, S. B.; Horn, D.; Schup-Magoffin, P. J.; Christman, K. L. Injectable Extracellular Matrix Derived Hydrogel Provides a Platform for Enhanced Retention and Delivery of a Heparin-Binding Growth Factor. Acta Biomater. 2012, 8, 3695– 3703, DOI: 10.1016/j.actbio.2012.06.03013https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38Xht1Kktb%252FO&md5=771332ec946c70753bfdd4e98acd2214Injectable extracellular matrix derived hydrogel provides a platform for enhanced retention and delivery of a heparin-binding growth factorSeif-Naraghi, Sonya B.; Horn, Dinah; Schup-Magoffin, Pamela J.; Christman, Karen L.Acta Biomaterialia (2012), 8 (10), 3695-3703CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Injectable hydrogels derived from the extracellular matrix (ECM) of decellularized tissues have recently emerged as scaffolds for tissue-engineering applications. Here, we introduce the potential for using a decellularized ECM-derived hydrogel for the improved delivery of heparin-binding growth factors. Immobilization of growth factors on a scaffold has been shown to increase their stability and activity. This can be done via chem. crosslinking, covalent bonding, or by incorporating natural or synthetic growth factor-binding domains similar to those found in vivo in sulfated glycosaminoglycans (GAGs). Many decellularized ECM-derived hydrogels retain native sulfated GAGs, and these materials may therefore provide an excellent delivery platform for heparin-binding growth factors. In this study, the sulfated GAG content of an ECM hydrogel derived from decellularized pericardial ECM was confirmed by Fourier transform IR spectroscopy and its ability to bind basic fibroblast growth factor (bFGF) was established. Delivery in the pericardial matrix hydrogel increased retention of bFGF both in vitro and in vivo in ischemic myocardium compared to delivery in collagen. In a rodent infarct model, intramyocardial injection of bFGF in pericardial matrix enhanced neovascularization by approx. 112% compared to delivery in collagen. Importantly, the newly formed vasculature was anastomosed with existing vasculature. Thus, the sulfated GAG content of the decellularized ECM hydrogel provides a platform for incorporation of heparin-binding growth factors for prolonged retention and delivery.
- 14Seo, Y.; Jung, Y.; Kim, S. H. Decellularized Heart ECM Hydrogel Using Supercritical Carbon Dioxide for Improved Angiogenesis. Acta Biomater. 2018, 67, 270– 281, DOI: 10.1016/j.actbio.2017.11.04614https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXitVaksbzO&md5=2ab93efb29e88bbe22dbfe56f54503d8Decellularized heart ECM hydrogel using supercritical carbon dioxide for improved angiogenesisSeo, Yoojin; Jung, Youngmee; Kim, Soo HyunActa Biomaterialia (2018), 67 (), 270-281CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Initial angiogenesis within the first 3days is crit. for healing ischemic diseases such as myocardial infarction. Recently, decellularized extracellular matrix (dECM) has been reported to provide tissue-derived ECM components and can be used as a scaffold for cell delivery for angiogenesis in tissue engineering. Decellularization by various detergents such as sodium dodecyl sulfate (SDS) and triton X-100 can remove the cell nuclei in tissue organs. However, this leads to ECM structure denaturation, decreased presence of various ECM proteins and cytokines, and loss of mech. properties. To overcome these limitations, in this study, we developed a supercrit. carbon dioxide and ethanol co-solvent (scCO2-EtOH) decellularization method, which is a detergent-free system that prevents ECM structure disruption and retains various angiogenic proteins in the heart dECM, and tested on rat heart tissues. The heart tissue was placed into the scCO2 reactor and decellularized at 37°C and 350bar. After scCO2-EtOH treatment, the effects were evaluated by DNA, collagen, and glycosaminoglycan (GAG) quantification and hematoxylin and eosin and immunofluorescence staining to det. the absence of nucleic acids and preservation of heart ECM components. Similar to the native group, the scCO2-EtOH group contained more ECM components such as collagen, GAGs, collagen I, laminin, and fibronectin and angiogenic factors including vascular endothelial growth factor, fibroblast growth factor, and platelet-derived growth factor and others in comparison to the detergent group. In addn., to est. angiogenesis of the dECM hydrogels, the neutralized dECM soln. was injected in a rat s.c. layer (n=6 in each group: collagen, scCO2-EOH, and detergent group), after which the soln. naturally formed gelation in the s.c. layer. After 3days, the gels were harvested and estd. by immunofluorescence staining and the ImageJ program for angiogenesis anal. Consequently, blood vessel formation and d. of vWF and α-SMA in the scCO2-EtOH group were significantly greater than that in the collagen group. Here we suggest that heart-derived decellularized extracellular matrix (dECM) with scCO2-EtOH treatment is a highly promising angiogenic material for healing in ischemic disease. Supercrit. carbon dioxide (scCO2) in a supercrit. phase has low viscosity and high diffusivity between gas and liq. properties and is known to be affordable, non-toxic, and eco-friendly. Therefore, scCO2 extn. technol. has been extensively used in com. and industrial fields. Recently, decellularized extracellular matrix (dECM) was applied to tissue engineering and regenerative medicine as a scaffold, therapeutic material, and bio-ink for 3D printing. Moreover, the general decellularization method using detergents has limitations including eliminating tissue-derived ECM components and disrupting their structures after decellularization. To overcome these limitations, heart tissues were treated with scCO2-EtOH for decellularization, resulting in preserving of tissue due to the various ECM and angiogenic factors derived. In addn., initiation of angiogenesis was highly induced even after 3days of injection.
- 15Bordbar, S.; Bakhshaiesh, N. L.; Khanmohammadi, M.; Sayahpour, F. A.; Alini, M.; Baghaban Eslaminejad, M. Production and Evaluation of Decellularized Extracellular Matrix Hydrogel for Cartilage Regeneration Derived from Knee Cartilage. J. Biomed. Mater. Res., Part A 2020, 108, 938– 946, DOI: 10.1002/jbm.a.3687115https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXjvVWksg%253D%253D&md5=8350496b436f9924c53b3b0d0f72a200Production and evaluation of decellularized extracellular matrix hydrogel for cartilage regeneration derived from knee cartilageBordbar, Sima; Lotfi Bakhshaiesh, Nasrin; Khanmohammadi, Mehdi; Sayahpour, Forough Azam; Alini, Mauro; Baghaban Eslaminejad, MohamadrezaJournal of Biomedical Materials Research, Part A (2020), 108 (4), 938-946CODEN: JBMRCH; ISSN:1549-3296. (John Wiley & Sons, Inc.)Cartilage tissue engineering is the interdisciplinary science that will help to improve cartilage afflictions, such as arthrosis, arthritis, or following joints traumatic injuries. In the present work, we developed an injectable hydrogel which derived from decellularized extracellular matrix of sheep cartilage. Successful decellularization was evaluated by measuring the DNA, glycosaminoglycans (GAG), collagen contents, and histol. analyses. There was a minor difference in GAG and collagen contents among natural cartilage and decellularized tissue as well as ultimate hydrogel. Rheol. anal. showed that the temp. and gelation time of prepd. hydrogel were 37°C and between 5 and 7 min, resp. Mech. properties evaluation indicated a storage modulus of 20 kPa. The results show that prepd. hydrogel possessed cell-friendly microenvironment as confirmed via calcein staining and MTT assay. Also, cells were able to proliferate which obsd. by H&E and alcian blue staining. Cell attachment and proliferation at the surface of the decellularized hydrogel was apparent by Scanning Electron Microscope (SEM) images and microphotographs. Furthermore, the cells embedded within the hydrogel were able to differentiate into chondrocyte with limited evidence of hypertrophy and osteogenesis in utilized cells which proved by SOX9, CoL2, ACAN, and also CoL1 and CoL10 gene expression levels. In summary, the results suggest that developed novel injectable hydrogel from decellularized cartilage could be utilized as a promising substrate for cartilage tissue engineering applications.
- 16Wolf, M. T.; Daly, K. A.; Brennan-Pierce, E. P.; Johnson, S. A.; Carruthers, C. A.; D’Amore, A.; Nagarkar, S. P.; Velankar, S. S.; Badylak, S. F. A Hydrogel Derived from Decellularized Dermal Extracellular Matrix. Biomaterials 2012, 33, 7028– 7038, DOI: 10.1016/j.biomaterials.2012.06.05116https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XhtVaktrfE&md5=0a4178709ca54cb1df8d5bfdddd1c2d1A hydrogel derived from decellularized dermal extracellular matrixWolf, Matthew T.; Daly, Kerry A.; Brennan-Pierce, Ellen P.; Johnson, Scott A.; Carruthers, Christopher A.; D'Amore, Antonio; Nagarkar, Shailesh P.; Velankar, Sachin S.; Badylak, Stephen F.Biomaterials (2012), 33 (29), 7028-7038CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)The ECM of mammalian tissues has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. Such scaffolds are prepd. in many forms including sheets, powders, and hydrogels. ECM hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the inherent bioactivity of native matrix. However, material properties of ECM hydrogels and the effect of these properties upon cell behavior are neither well understood nor controlled. The objective of this study was to prep. and det. the structure, mechanics, and the cell response in vitro and in vivo of ECM hydrogels prepd. from decellularized porcine dermis and urinary bladder tissues. Dermal ECM hydrogels were characterized by a more dense fiber architecture and greater mech. integrity than urinary bladder ECM hydrogels, and showed a dose dependent increase in mech. properties with ECM concn. In vitro, dermal ECM hydrogels supported greater C2C12 myoblast fusion, and less fibroblast infiltration and less fibroblast mediated hydrogel contraction than urinary bladder ECM hydrogels. Both hydrogels were rapidly infiltrated by host cells, primarily macrophages, when implanted in a rat abdominal wall defect. Both ECM hydrogels degraded by 35 days in vivo, but UBM hydrogels degraded more quickly, and with greater amts. of myogenesis than dermal ECM. These results show that ECM hydrogel properties can be varied and partially controlled by the scaffold tissue source, and that these properties can markedly affect cell behavior.
- 17Badylak, S. F.; Freytes, D. O.; Gilbert, T. W. Extracellular Matrix as a Biological Scaffold Material: Structure and Function. Acta Biomater. 2009, 5, 1– 13, DOI: 10.1016/j.actbio.2008.09.01317https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXhsVamsbc%253D&md5=fd27594c129e4f996c49f69a233dbffcExtracellular matrix as a biological scaffold material: structure and functionBadylak, Stephen F.; Freytes, Donald O.; Gilbert, Thomas W.Acta Biomaterialia (2009), 5 (1), 1-13CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)A review. Biol. scaffold materials derived from the extracellular matrix (ECM) of intact mammalian tissues have been successfully used in a variety of tissue engineering/regenerative, medicine applications both in preclin. studies and in clin. applications. Although it is recognized that the materials have constructive remodeling properties, the mechanisms by which functional tissue restoration is achieved are not well understood. There is evidence to support essential roles for both the structural and functional characteristics of the biol. scaffold materials. This paper provides an overview of the compn. and structure of selected ECM scaffold materials, the effects of manufg. methods upon the structural properties and resulting mech. behavior of the scaffold materials, and the in vivo degrdn. and remodeling of ECM scaffolds with an emphasis on tissue function.
- 18Xu, J.; Fang, H.; Zheng, S.; Li, L.; Jiao, Z.; Wang, H.; Nie, Y.; Liu, T.; Song, K. A Biological Functional Hybrid Scaffold Based on Decellularized Extracellular Matrix/Gelatin/Chitosan with High Biocompatibility and Antibacterial Activity for Skin Tissue Engineering. Int. J. Biol. Macromol. 2021, 187, 840– 849, DOI: 10.1016/j.ijbiomac.2021.07.16218https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhslGis7%252FF&md5=bdc31c02e4ec3773c784ee626ce31a9eA biological functional hybrid scaffold based on decellularized extracellular matrix/gelatin/chitosan with high biocompatibility and antibacterial activity for skin tissue engineeringXu, Jie; Fang, Huan; Zheng, Shuangshuang; Li, Liying; Jiao, Zeren; Wang, Hong; Nie, Yi; Liu, Tianqing; Song, KedongInternational Journal of Biological Macromolecules (2021), 187 (), 840-849CODEN: IJBMDR; ISSN:0141-8130. (Elsevier B.V.)Nowadays, decellularized extracellular matrix (dECM) has received widespread attention due to its diversity in providing the unique structural and functional components to support cell growth, and finding material with good biocompatibility and anti-infection capability for skin tissue engineering is still a challenge. In this study, a novel dECM/Gel/CS scaffold with appropriate mech. strength, good antibacterial activity and high biocompatibility was prepd. using a one-pot method. The results showed that the immune components such as cells and DNA (about 98.1%) were successfully removed from the porcine skin tissue. The dECM/Gel/CS scaffolds exhibited an interconnected pore structure and had a high porosity (>90%) to promote cell growth. Moreover, the appropriate elastic modulus (≥482.17 kPa) and degradability (≥80.04% for 15 days) of the scaffolds offered stout "houses" for cell proliferation and suitable degrdn. rate to match the new tissue formation in skin tissue engineering. Furthermore, the addn. of chitosan endowed the scaffold with good antibacterial activity, water and protein absorption capacity to avoid wound infection, and maintain the moisture and nutrition balance. In vitro cytocompatibility studies showed that the presence of dECM effectively enhanced the cell proliferation. Overall, the advanced dECM/Gel/CS scaffold has considerable potential to be applied in skin tissue engineering.
- 19Su, Z.; Ma, H.; Wu, Z.; Zeng, H.; Li, Z.; Wang, Y.; Liu, G.; Xu, B.; Lin, Y.; Zhang, P.; Wei, X. Enhancement of Skin Wound Healing with Decellularized Scaffolds Loaded with Hyaluronic Acid and Epidermal Growth Factor. Mater. Sci. Eng. C 2014, 44, 440– 448, DOI: 10.1016/j.msec.2014.07.03919https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhs1KgtrzI&md5=416018c803a3f51d58443edf981f96bbEnhancement of skin wound healing with decellularized scaffolds loaded with hyaluronic acid and epidermal growth factorSu, Zhongchun; Ma, Huan; Wu, Zhengzheng; Zeng, Huilan; Li, Zhizhong; Wang, Yuechun; Liu, Gexiu; Xu, Bin; Lin, Yongliang; Zhang, Peng; Wei, XingMaterials Science & Engineering, C: Materials for Biological Applications (2014), 44 (), 440-448CODEN: MSCEEE; ISSN:0928-4931. (Elsevier B.V.)Current therapy for skin wound healing still relies on skin transplantation. Many studies were done to try to find out ways to replace skin transplantation, but there is still no effective alternative therapy. In this study, decellularized scaffolds were prepd. from pig peritoneum by a series of phys. and chem. treatments, and scaffolds loaded with hyaluronic acid (HA) and epidermal growth factor (EGF) were tested for their effect on wound healing. MTT assay showed that EGF increased NIH3T3 cell viability and confirmed that EGF used in this study was biol. active in vitro. Scanning electron microscope (SEM) showed that HA stably attached to scaffolds even after soaking in PBS for 48 h. ELISA assay showed that HA increased the adsorption of EGF to scaffolds and sustained the release of EGF from scaffolds. Animal study showed that the wounds covered with scaffolds contg. HA and EGF recovered best among all 4 groups and had wound healing rates of 49.86%, 70.94% and 87.41% resp. for days 10, 15 and 20 post-surgery compared to scaffolds alone with wound healing rates of 29.26%, 42.80% and 70.14%. In addn., the wounds covered with scaffolds contg. EGF alone were smaller than no EGF scaffolds on days 10, 15 and 20 post-surgery. Hematoxylin-Eosin (HE) staining confirmed these results by showing that on days 10, 15 and 20 post-surgery, the thicker epidermis and dermis layers were obsd. in the wounds covered with scaffolds contg. HA and EGF than scaffolds alone. In addn., the thicker epidermis and dermis layers were also obsd. in the wounds covered with scaffolds contg. EGF than scaffolds alone. Skin appendages were obsd. on day 20 only in the wound covered with scaffolds contg. HA and EGF. These results demonstrate that the scaffolds contg. HA and EGF can enhance wound healing.
- 20Chen, W.; Xu, Y.; Li, Y.; Jia, L.; Mo, X.; Jiang, G.; Zhou, G. 3D Printing Electrospinning Fiber-Reinforced Decellularized Extracellular Matrix for Cartilage Regeneration. Chem. Eng. J. 2020, 382, 122986 DOI: 10.1016/j.cej.2019.12298620https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvFejtrzL&md5=1a35b80756b9832a8a258796ab71f0d43D printing electrospinning fiber-reinforced decellularized extracellular matrix for cartilage regenerationChen, Weiming; Xu, Yong; Li, Yaqiang; Jia, Litao; Mo, Xiumei; Jiang, Gening; Zhou, GuangdongChemical Engineering Journal (Amsterdam, Netherlands) (2020), 382 (), 122986CODEN: CMEJAJ; ISSN:1385-8947. (Elsevier B.V.)Cartilage decellularized matrix (CDM) is considered a promising biomaterial for fabricating cartilage tissue engineering scaffolds. An ideal CDM-based scaffold should possess customizable 3D shape for complex tissue regeneration and proper pore size for cell infiltration, as well as provide mech. support for cell growth. 3D printing is an efficiently technique for prepg. customizable 3D scaffolds, however, fabricating CDM-based 3D-printed scaffolds with customizable shapes, proper pore structure and satisfactory mech. properties remains a challenge. In the current study, to achieve customizable CDM-based 3D scaffolds, CDM was successfully processed into inks suitable for 3D printing. Further, the poor mechanics of CDM-based scaffolds were significantly improved by adding electrospinning fiber into the CDM-based inks for 3D printing. Importantly, the 3D-printed electrospinning fiber-reinforced CDM-based scaffold presented good biocompatibility and can enhance repair articular cartilage defects in rabbits. The current study provides a novel strategy for printing electrospinning fiber-reinforced CDM-based scaffolds for tissue regeneration.
- 21Wang, Z.; Li, Z.; Li, Z.; Wu, B.; Liu, Y.; Wu, W. Cartilaginous Extracellular Matrix Derived from Decellularized Chondrocyte Sheets for the Reconstruction of Osteochondral Defects in Rabbits. Acta Biomater. 2018, 81, 129– 145, DOI: 10.1016/j.actbio.2018.10.00521https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhvFSqu7jK&md5=7b2448e86db36a983a1cf4c124800d75Cartilaginous extracellular matrix derived from decellularized chondrocyte sheets for the reconstruction of osteochondral defects in rabbitsWang, Zhifa; Li, Zhiye; Li, Zhijin; Wu, Buling; Liu, Yanpu; Wu, WeiActa Biomaterialia (2018), 81 (), 129-145CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Cartilaginous extracellular matrix (ECM) materials derived from decellularized native articular cartilage are widely used in cartilage regeneration. However, it is difficult for endogenous cells to migrate into ECM derived from native cartilage owing to its nonporous structure and dense nature. We cultured chondrocytes harvested from the auricular cartilage of 4-wk-old New Zealand rabbits and enabled them to form cell sheets. In vitro microstructural examn. and mech. tests demonstrated that 1% SDS not only removed chondrocytes completely but also maintained the native architecture and compn. of ECM, thus avoiding the use of high-concn. SDS. In conclusion, our results suggested that the chondrocyte sheets decellularized with 1% SDS preserved the integrity and bioactivity, which favored cell recruitment and enabled osteochondral regeneration in the knee joints of rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation. Our results suggested that cartilaginous ECM favored cell recruitment and enabled osteochondral regeneration in rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation. SDS 1% adequately decellularized the chondrocytes in cell sheets, whereas it maintained the native architecture and compn. of ECM, thereby avoiding the use of high-concn. SDS and providing a new way to acquire cartilaginous ECM.
- 22Han, T. T. Y.; Toutounji, S.; Amsden, B. G.; Flynn, L. E. Adipose-Derived Stromal Cells Mediate in Vivo Adipogenesis, Angiogenesis and Inflammation in Decellularized Adipose Tissue Bioscaffolds. Biomaterials 2015, 72, 125– 137, DOI: 10.1016/j.biomaterials.2015.08.05322https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXhsVaqsL7N&md5=6a74a992d2da07c5e4a667862c9a6f9dAdipose-derived stromal cells mediate in vivo adipogenesis, angiogenesis and inflammation in decellularized adipose tissue bioscaffoldsHan, Tim Tian Y.; Toutounji, Sandra; Amsden, Brian G.; Flynn, Lauren E.Biomaterials (2015), 72 (), 125-137CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)Decellularized adipose tissue (DAT) has shown promise as an adipogenic bioscaffold for soft tissue augmentation and reconstruction. The objective of the current study was to investigate the effects of allogeneic adipose-derived stem/stromal cells (ASCs) on in vivo fat regeneration in DAT bioscaffolds using an immunocompetent rat model. ASC seeding significantly enhanced angiogenesis and adipogenesis, with cell tracking studies indicating that the newly-forming tissues were host-derived. Incorporating ASCs also mediated the inflammatory response and promoted a more constructive macrophage phenotype. A fraction of the CD163+ macrophages in the implants expressed adipogenic markers, with higher levels of this "adipocyte-like" phenotype in proximity to the developing adipose tissues. Our results indicate that the combination of ASCs and adipose extracellular matrix (ECM) provides an inductive microenvironment for adipose regeneration mediated by infiltrating host cell populations. The DAT scaffolds are a useful tissue-specific model system for investigating the mechanisms of in vivo adipogenesis that may help to develop a better understanding of this complex process in the context of both regeneration and disease. Overall, combining adipose-derived matrixes with ASCs is a highly promising approach for the in situ regeneration of host-derived adipose tissue.
- 23Han, W.; Singh, N. K.; Kim, J. J.; Kim, H.; Kim, B. S.; Park, J. Y.; Jang, J.; Cho, D. W. Directed Differential Behaviors of Multipotent Adult Stem Cells from Decellularized Tissue/Organ Extracellular Matrix Bioinks. Biomaterials 2019, 224, 119496 DOI: 10.1016/j.biomaterials.2019.11949623https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhvVamt7fO&md5=a3eb11fd9fa07b402b08429ec61ef19aDirected differential behaviors of multipotent adult stem cells from decellularized tissue/organ extracellular matrix bioinksHan, Wonil; Singh, Narendra K.; Kim, Joeng Ju; Kim, Hyeonji; Kim, Byoung Soo; Park, Ju Young; Jang, Jinah; Cho, Dong-WooBiomaterials (2019), 224 (), 119496CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)The decellularized tissue/organ extracellular matrix (dECM) is a naturally derived biomaterial that inherits various functional components from the native tissue or organ. Recently, various kinds of tissue/organ dECM bioinks capable of encapsulating cells, combined with 3D cell printing, have enabled remarkable progress in tissue engineering and regenerative medicine. However, the way in which the dECM component compns. of each tissue of different origins interact with cells and dictate tissue-specific cell behavior in the 3D microenvironment remains mostly unknown. To address this issue, in-depth differential proteomic analyses of four porcine dECMs were performed. Specifically, the differential variations of matrisome protein compn. in each decellularized tissue type were also uncovered, which can play a significant role by affecting the resident cells in specific tissues. Furthermore, microarray analyses of human bone marrow mesenchymal stem cells (hBMMSCs) printed with various dECM bioinks were conducted to reveal the effect of compositional variations in a tissue-specific manner at the cellular level depending on the multipotency of MSCs. Through whole transcriptome anal., differential expression patterns of genes were obsd. in a tissue-specific manner, and this research provides strong evidence of the tissue-specific functionalities of dECM bioinks.
- 24Zhou, H.; Chen, R.; Wang, J.; Lu, J.; Yu, T.; Wu, X.; Xu, S.; Li, Z.; Jie, C.; Cao, R.; Yang, Y.; Li, Y.; Meng, D. Biphasic Fish Collagen Scaffold for Osteochondral Regeneration. Mater. Des. 2020, 195, 108947 DOI: 10.1016/j.matdes.2020.10894724https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXhsF2qsbzM&md5=5107bf9dd64bd9cb3aa0ba8429ecbb88Biphasic fish collagen scaffold for osteochondral regenerationZhou, Haichao; Chen, Ru; Wang, Jinpeng; Lu, Jia; Yu, Tao; Wu, Xinbo; Xu, Shaochen; Li, Zihua; Jie, Chen; Cao, Runfeng; Yang, Yunfeng; Li, Yaqiang; Meng, DepengMaterials & Design (2020), 195 (), 108947CODEN: MADSD2; ISSN:0264-1275. (Elsevier Ltd.)Developing a biphasic scaffold that can concurrently regenerate both cartilage and bone of osteochondral defects (OCDs) is a challenge. Fish collagen (FC) is currently considered an alternative scaffold to mammalian collagen (MC) due to its safety, accessibility, lower price, and similar biol. properties compared to those of MC. Here, we developed bilayer FC-based composite scaffolds with different components and pore sizes to modulate the differentiation fate of bone marrow stem cells (BMSCs): chondroitin sulfate-incorporated FC scaffolds (FC-CS) with small pores (approx. 128μm) as the top layer and hydroxyapatite-incorporated FC scaffolds (FC-HA) with larger pores (approx. 326μm) as the bottom layer. Both the FC-CS and FC-HA scaffolds possessed good cytocompatibility, excellent water absorption, suitable biodegradability and high cell seeding efficiency. The in vitro results indicated that FC-CS and FC-HA promote chondrogenesis and osteogenesis of BMSCs, resp., as validated by gene expression and histol. examn. Furthermore, compared to the empty group in a rabbit OCD model, the bilayer scaffold significantly induced simultaneous regeneration of cartilage and subchondral bone after 6 and 12 wk of implantation, which was confirmed by gross, histol., and microcomputed tomog. images. Our findings demonstrated that the FC-based bilayer scaffold is a promising scaffold for the repair of OCD.
- 25Kirsner, R. S.; Margolis, D. J.; Baldursson, B. T.; Petursdottir, K.; Davidsson, O. B.; Weir, D.; Lantis, J. C. Fish Skin Grafts Compared to Human Amnion/Chorion Membrane Allografts: A Double-Blind, Prospective, Randomized Clinical Trial of Acute Wound Healing. Wound Repair Regener. 2020, 28, 75– 80, DOI: 10.1111/wrr.1276125https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3Mrns1ShsA%253D%253D&md5=90d00ef9004c0d03ed7e30810307702dFish skin grafts compared to human amnion/chorion membrane allografts: A double-blind, prospective, randomized clinical trial of acute wound healingKirsner Robert S; Margolis David J; Margolis David J; Baldursson Baldur T; Petursdottir Kristin; Davidsson Olafur B; Weir Dot; Lantis John C 2ndWound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society (2020), 28 (1), 75-80 ISSN:.Chronic, nonhealing wounds consume a great deal of healthcare resources and are a major public health problem, associated with high morbidity and significant economic costs. Skin grafts are commonly used to facilitate wound closure. The grafts can come from the patient's own skin (autograft), a human donor (allograft), or from a different species (xenograft). A fish skin xenograft from cold-water fish (Atlantic cod, Gadus morhua) is a relatively recent option that shows promising preclinical and clinical results in wound healing. Chronic wounds vary greatly in etiology and nature, requiring large cohorts for effective comparison between therapeutic alternatives. In this study, we attempted to imitate the status of a freshly debrided chronic wound by creating acute full-thickness wounds, 4 mm in diameter, on healthy volunteers to compare two materials frequently used to treat chronic wounds: fish skin and dHACM. The purpose is to give an indication of the efficacy of the two therapeutic alternatives in the treatment of chronic wounds in a simple, standardized, randomized, controlled, double-blind study. All volunteers were given two identical punch biopsy wounds, one of which was treated with a fish skin graft and the other with dehydrated human amnion/chorion membrane allograft (dHACM). In the study, 170 wounds were treated (85 wounds per group). The primary endpoint was defined as time to heal (full epithelialization) by blinded assessment at days 14, 18, 21, 25, and 28. The superiority hypothesis was that the fish skin grafts would heal the wounds faster than the dHACM. To evaluate the superiority hypothesis, a mixed Cox proportional hazard model was used. Wounds treated with fish skin healed significantly faster (hazard ratio 2.37; 95% confidence interval: (1.75-3.22; p = 0.0014) compared with wounds treated with dHACM. The results show that acute biopsy wounds treated with fish skin grafts heal faster than wounds treated with dHACM.
- 26Govindharaj, M.; Roopavath, U. K.; Rath, S. N. Valorization of Discarded Marine Eel Fish Skin for Collagen Extraction as a 3D Printable Blue Biomaterial for Tissue Engineering. J. Cleaner Prod. 2019, 230, 412– 419, DOI: 10.1016/j.jclepro.2019.05.08226https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtVKgurvP&md5=057db7abdea5b3a139fab2790428ea84Valorization of discarded Marine Eel fish skin for collagen extraction as a 3D printable blue biomaterial for tissue engineeringGovindharaj, Mano; Roopavath, Uday Kiran; Rath, Subha NarayanJournal of Cleaner Production (2019), 230 (), 412-419CODEN: JCROE8; ISSN:0959-6526. (Elsevier Ltd.)Discarded marine Eel fish skin has essential properties of biomaterials for potential use in tissue engineering application. Processing and prepn. of eel fish for edible purpose requires the removal of skin due to its thick size, which is treated as a waste. A huge amt. of Eel skin is dumped as a waste material which leads to marine environmental pollution. To overcome this issue, we have isolated collagen from the discarded marine Eel skin as a potential blue biomaterial. Further, the isolated collagen was incorporated into alginate hydrogel to fabricate scaffolds using extrusion-based 3D printing technol. Swelling, degrdn. and biocompatibility were evaluated for lyophilized scaffolds. Biocompatibility studies were performed on hUMSCs (Human Umbilical cord Derived Mesenchymal Stem Cells) by live/dead staining using FDA (fluorescein diacetate)/PI (Propidium Iodide). The quant. evaluation of metabolic activity was performed using Alamar Blue (AB) dye redn. assay. All the hydrogels with collagen show enhanced metabolic activity and cell proliferation compared to alginate hydrogels without collagen. The utilization of Eel skin derived collagen for 3D printing application was not yet reported. Moreover, sustainable utilization of renewable marine Eel skin discard as a novel blue biomaterial is of immense value due to its low cost and has great potential for further tissue engineering applications.
- 27Blanco, M.; Vázquez, J. A.; Pérez-Martín, R. I.; Sotelo, C. G. Collagen Extraction Optimization from the Skin of the Small-Spotted Catshark (S. Canicula) by Response Surface Methodology. Mar. Drugs 2019, 17, 40 DOI: 10.3390/md1701004027https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtFGmtrzJ&md5=76da1a9f390bb5c4fba379151e0dcf2fCollagen extraction optimization from the skin of the small-spotted catshark (S. canicula) by response surface methodologyBlanco, Maria; Vazquez, Jose Antonio; Perez-Martin, Ricardo I.; Sotelo, Carmen G.Marine Drugs (2019), 17 (1), 40CODEN: MDARE6; ISSN:1660-3397. (MDPI AG)The small-spotted catshark is one of the most abundant elasmobranchs in the Northeastern Atlantic Ocean. Although its landings are devoted for human consumption, in general this species has low com. value with high discard rates, reaching 100% in some European fisheries. The eduction of post-harvest losses (discards and byproducts) by promotion of a full use of fishing captures is one of the main goals of EU fishing policies. As marine collagens are increasingly used as alternatives to mammalian collagens for cosmetics, tissue engineering, etc., fish skins represent an excellent and abundant source for obtaining this biomol. The aim of this study was to analyze the influence of chem. treatment concn., temp. and time on the extractability of skin collagen from this species. Two exptl. designs, one for each of the main stages of the process, were performed by means of Response Surface Methodol. (RSM). The combined effect of NaOH concn., time and temp. on the amt. of collagen recovered in the first stage of the collagen extn. procedure was studied. Then, skins treated under optimal NaOH conditions were subjected to a second exptl. design, to study the combined effect of AcOH concn., time and temp. on the collagen recovery by means of yield, amino acid content and SDS-PAGE characterization. Values of independent variables maximizing collagen recovery were 4°C, 2 h and 0.1 M NaOH (pre-treatment) and 25°C, 34 h and 1 M AcOH (collagen extn.).
- 28Lim, Y.; Ok, Y.-J.; Hwang, S.; Kwak, J.; Yoon, S. Marine Collagen as A Promising Biomaterial For. Mar. Drugs 2019, 17, 467 DOI: 10.3390/md1708046728https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXks1Oks7g%253D&md5=e9dd3e6d1288e11100187bd1ec534c35Marine collagen as a promising biomaterial for biomedical applicationsLim, Ye-Seon; Ok, Ye-Jin; Hwang, Seon-Yeong; Kwak, Jong-Young; Yoon, SikMarine Drugs (2019), 17 (8), 467CODEN: MDARE6; ISSN:1660-3397. (MDPI AG)This review focuses on the expanding role of marine collagen (MC)-based scaffolds for biomedical applications. A scaffold-a three-dimensional (3D) structure fabricated from biomaterials-is a key supporting element for cell attachment, growth, and maintenance in 3D cell culture and tissue engineering. The mech. and biol. properties of the scaffolds influence cell morphol., behavior, and function. MC, collagen derived from marine organisms, offers advantages over mammalian collagen due to its biocompatibility, biodegradability, easy extractability, water soly., safety, low immunogenicity, and low prodn. costs. In recent years, the use of MC as an increasingly valuable scaffold biomaterial has drawn considerable attention from biomedical researchers. The characteristics, isolation, phys., and biochem. properties of MC are discussed as an understanding of MC in optimizing the subsequent modification and the chemistries behind important tissue engineering applications. The latest technologies behind scaffold processing are assessed and the biomedical applications of MC and MC-based scaffolds, including tissue engineering and regeneration, wound dressing, drug delivery, and therapeutic approach for diseases, esp. those assocd. with metabolic disturbances such as obesity and diabetes, are discussed. Despite all the challenges, MC holds great promise as a biomaterial for developing medical products and therapeutics.
- 29Girardeau-Hubert, S.; Lynch, B.; Zuttion, F.; Label, R.; Rayee, C.; Brizion, S.; Ricois, S.; Martinez, A.; Park, E.; Kim, C.; Marinho, P. A.; Shim, J. H.; Jin, S.; Rielland, M.; Soeur, J. Impact of Microstructure on Cell Behavior and Tissue Mechanics in Collagen and Dermal Decellularized Extra-Cellular Matrices. Acta Biomater. 2022, 143, 100– 114, DOI: 10.1016/j.actbio.2022.02.03529https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XhsVSgtLnL&md5=3f9072612ae4d39007df8771f8b46986Impact of microstructure on cell behavior and tissue mechanics in collagen and dermal decellularized extra-cellular matricesGirardeau-Hubert, Sarah; Lynch, Barbara; Zuttion, Francesca; Label, Rabab; Rayee, Chrystelle; Brizion, Sebastien; Ricois, Sylvie; Martinez, Anthony; Park, Eunhye; Kim, Changhwan; Marinho, Paulo Andre; Shim, Jin-Hyung; Jin, Songwan; Rielland, Maite; Soeur, JeremieActa Biomaterialia (2022), 143 (), 100-114CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Skin models are used for many applications such as research and development or grafting. Unfortunately, most lack a proper microenvironment producing poor mech. properties and inaccurate extra-cellular matrix compn. and organization. In this report we focused on mech. properties, extra-cellular matrix organization and cell interactions in human skin samples reconstructed with pure collagen or dermal decellularized extra-cellular matrixes (S-dECM) and compared them to native human skin. We found that Full-thickness S-dECM samples presented stiffness two times higher than collagen gel and similar to ex vivo human skin, and proved for the first time that keratinocytes also impact dermal mech. properties. This was correlated with larger fibers in S-dECM matrixes compared to collagen samples and with a differential expression of F-actin, vinculin and tenascin C between S-dECM and collagen samples. This is clear proof of the microenvironments impact on cell behaviors and mech. properties. In vitro skin models have been used for a long time for clin. applications or in vitro knowledge and evaluation studies. However, most lack a proper microenvironment producing a poor combination of mech. properties and appropriate biol. outcomes, partly due to inaccurate extra-cellular matrix (ECM) compn. and organization. This can lead to limited predictivity and weakness of skin substitutes after grafting. This study shows, for the first time, the importance of a complex and rich microenvironment on cell behaviors, matrix macro- and micro-organization and mech. properties. The increased compn. and organization complexity of dermal skin decellularized extra-cellular matrix populated with differentiated cells produces in vitro skin models closer to native human skin physiol.
- 30Chameettachal, S.; Sasikumar, S.; Sethi, S.; Sriya, Y.; Pati, F. Tissue/Organ-Derived Bioink Formulation for 3D Bioprinting. J. 3D Print. Med. 2019, 3, 39– 54, DOI: 10.2217/3dp-2018-002430https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXkslamt7Y%253D&md5=c70ce9cf2f9c97eaf43066f5b77ec00dTissue/organ-derived bioink formulation for 3D bioprintingChameettachal, Shibu; Sasikumar, Shyama; Sethi, Soumya; Sriya, Yeleswarapu; Pati, FalguniJournal of 3D Printing in Medicine (2019), 3 (1), 39-54CODEN: JDPMAT; ISSN:2059-4763. (Future Medicine Ltd.)Tissue/organ-derived bioink formulations open up new avenues in 3D bioprinting research with the potential to create functional tissue or organs. Printing of tissue construct largely depends on material properties, as it needs to be fabricated in an aq. environment while encapsulating living cells. The decellularized extracellular matrix bioinks proved to be a potential option for functional tissue development in vivo and as an alternative to chem. cross-linked bioinks. However, certain limitations such as printability and limited mech. strength need to be addressed for enhancing their widespread applications. By drawing knowledge from the existing literature, emphasis has been given in this review to the development of decellularized extracellular matrix bioinks and their applications in printing functional tissue constructs.
- 31Reing, J. E.; Brown, B. N.; Daly, K. A.; Freund, J. M.; Gilbert, T. W.; Hsiong, S. X.; Huber, A.; Kullas, K. E.; Tottey, S.; Wolf, M. T.; Badylak, S. F. The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds. Biomaterials 2010, 31, 8626– 8633, DOI: 10.1016/j.biomaterials.2010.07.08331https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXht1ehtLnP&md5=5f2f35e650c7a0b64fdfadc67becd2c7The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffoldsReing, Janet E.; Brown, Bryan N.; Daly, Kerry A.; Freund, John M.; Gilbert, Thomas W.; Hsiong, Susan X.; Huber, Alexander; Kullas, Karen E.; Tottey, Stephen; Wolf, Matthew T.; Badylak, Stephen F.Biomaterials (2010), 31 (33), 8626-8633CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)Biol. materials from various species and tissues are commonly used as surgical meshes or scaffolds for tissue reconstruction. Extracellular matrix (ECM) represents the secreted product of the cells comprising each tissue and organ, and therefore provides a unique biol. material for selected regenerative medicine applications. Minimal disruption of ECM ultrastructure and content during tissue processing is typically desirable. The objective of this study was to systematically evaluate effects of commonly used tissue processing steps upon porcine dermal ECM scaffold compn., mech. properties, and cytocompatibility. Processing steps evaluated included liming and hot water sanitation, trypsin/SDS/TritonX-100 decellularization, and trypsin/TritonX-100 decellularization. Liming decreased the growth factor and glycosaminoglycan content, the mech. strength, and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for all). Hot water sanitation treatment decreased only the growth factor content of the ECM (p ≤ 0.05). Trypsin/SDS/TritonX-100 decellularization decreased the growth factor content and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). Trypsin/Triton X-100 decellularization also decreased the growth factor content of the ECM but increased the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). The authors conclude that processing steps evaluated in the present study affect content, mech. strength, and/or cytocompatibility of the resultant porcine dermal ECM, and therefore care must be taken in choosing appropriate processing steps to maintain the beneficial effects of ECM in biol. scaffolds.
- 32Zhang, Q.; Johnson, J. A.; Dunne, L. W.; Chen, Y.; Iyyanki, T.; Wu, Y.; Chang, E. I.; Branch-Brooks, C. D.; Robb, G. L.; Butler, C. E. Decellularized Skin/Adipose Tissue Flap Matrix for Engineering Vascularized Composite Soft Tissue Flaps. Acta Biomater. 2016, 35, 166– 184, DOI: 10.1016/j.actbio.2016.02.01732https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28Xis1CqtLY%253D&md5=85c5917bc2329157c22a3b88025b0cfdDecellularized skin/adipose tissue flap matrix for engineering vascularized composite soft tissue flapsZhang, Qixu; Johnson, Joshua A.; Dunne, Lina W.; Chen, Youbai; Iyyanki, Tejaswi; Wu, Yewen; Chang, Edward I.; Branch-Brooks, Cynthia D.; Robb, Geoffrey L.; Butler, Charles E.Acta Biomaterialia (2016), 35 (), 166-184CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Using a perfusion decellularization protocol, we developed a decellularized skin/adipose tissue flap (DSAF) comprising extracellular matrix (ECM) and intact vasculature. Our DSAF had a dominant vascular pedicle, microcirculatory vascularity, and a sensory nerve network and retained three-dimensional (3D) nanofibrous structures well. DSAF, which was composed of collagen and laminin with well-preserved growth factors (e.g., vascular endothelial growth factor, basic fibroblast growth factor), was successfully repopulated with human adipose-derived stem cells (hASCs) and human umbilical vein endothelial cells (HUVECs), which integrated with DSAF and formed 3D aggregates and vessel-like structures in vitro. We used microsurgery techniques to re-anastomose the recellularized DSAF into nude rats. In vivo, the engineered flap construct underwent neovascularization and constructive remodeling, which was characterized by the predominant infiltration of M2 macrophages and significant adipose tissue formation at 3 mo postoperatively. Our results indicate that DSAF co-cultured with hASCs and HUVECs is a promising platform for vascularized soft tissue flap engineering. This platform is not limited by the flap size, as the entire construct can be immediately perfused by the recellularized vascular network following simple re-integration into the host using conventional microsurgical techniques. Significant soft tissue loss resulting from traumatic injury or tumor resection often requires surgical reconstruction using autologous soft tissue flaps. However, the limited availability of qual. autologous flaps as well as the donor site morbidity significantly limits this approach. Engineered soft tissue flap grafts may offer a clin. relevant alternative to the autologous flap tissue. In this study, we engineered vascularized soft tissue free flap by using skin/adipose flap extracellular matrix scaffold (DSAF) in combination with multiple types of human cells. Following vascular reanastomosis in the recipient site, the engineered products successful regenerated large-scale fat tissue in vivo. This approach may provide a translatable platform for composite soft tissue free flap engineering for microsurgical reconstruction.
- 33Liu, D.; Wei, G.; Li, T.; Hu, J.; Lu, N.; Regenstein, J. M.; Zhou, P. Effects of Alkaline Pretreatments and Acid Extraction Conditions on the Acid-Soluble Collagen from Grass Carp (Ctenopharyngodon Idella) Skin. Food Chem. 2015, 172, 836– 843, DOI: 10.1016/j.foodchem.2014.09.14733https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhslajtb7L&md5=423b7904b4e4a78a130acb3f00793958Effects of alkaline pretreatments and acid extraction conditions on the acid-soluble collagen from grass carp (Ctenopharyngodon idella) skinLiu, Dasong; Wei, Guanmian; Li, Tiancheng; Hu, Jinhua; Lu, Naiyan; Regenstein, Joe M.; Zhou, PengFood Chemistry (2015), 172 (), 836-843CODEN: FOCHDJ; ISSN:0308-8146. (Elsevier Ltd.)This study investigated the effects of alk. pretreatments and acid extn. conditions on the prodn. of acid-sol. collagen (ASC) from grass carp skin. For alk. pretreatment, 0.05 and 0.1 M NaOH removed non-collagenous proteins without significant loss of ASC at 4, 10, 15 and 20 °C; while 0.2 and 0.5 M NaOH caused significant loss of ASC, and 0.5 M NaOH caused structural modification of ASC at 15 and 20 °C. For acid extn. at 4, 10, 15 and 20 °C, ASC was partly extd. by 0.1 and 0.2 M acetic acid, while 0.5 and 1.0 M acetic acid resulted in almost complete extn. The processing conditions involving 0.05-0.1 M NaOH for pretreatment, 0.5 M acetic acid for extn. and 4-20 °C for both pretreatment and extn., produced ASC with the structural integrity being well maintained and hence were recommended to prep. ASC from grass carp skin in practical application.
- 34Meng, D.; Tanaka, H.; Kobayashi, T.; Hatayama, H.; Zhang, X.; Ura, K.; Yunoki, S.; Takagi, Y. The Effect of Alkaline Pretreatment on the Biochemical Characteristics and Fibril-Forming Abilities of Types I and II Collagen Extracted from Bester Sturgeon by-Products. Int. J. Biol. Macromol. 2019, 131, 572– 580, DOI: 10.1016/j.ijbiomac.2019.03.09134https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXlvVWht7s%253D&md5=41b12c2b9764573c1ddc31372f3922aeThe effect of alkaline pretreatment on the biochemical characteristics and fibril-forming abilities of types I and II collagen extracted from bester sturgeon by-productsMeng, Dawei; Tanaka, Hiroyuki; Kobayashi, Taishi; Hatayama, Hirosuke; Zhang, Xi; Ura, Kazuhiro; Yunoki, Shunji; Takagi, YasuakiInternational Journal of Biological Macromolecules (2019), 131 (), 572-580CODEN: IJBMDR; ISSN:0141-8130. (Elsevier B.V.)Non-mammalian collagens have attracted increasing attention for industrial and biomedical use. We have therefore evaluated extn. conditions and the biochem. properties of collagens from aquacultured sturgeon. Pepsin-sol. type I and type II collagen were resp. extd. from the skin and notochord of bester sturgeon byproducts, with yields of 63.9 ± 0.19% and 35.5 ± 0.68%. Collagen extn. efficiency was improved by an alk. pretreatment of the skin and notochord (fewer extn. cycles were required), but the final yields decreased to 56.2 ± 0.84% for type I and 31.8 ± 1.13% for type II. Alk. pretreatment did not affect the thermal stability or triple-helical structure of both types of collagen. Types I and II collagen formed re-assembled fibril structures in vitro, under different conditions. Alk. pretreatment slowed down the formation of type I collagen fibrils and specifically inhibited the formation of thick fibril-bundle structures. In contrast, alk. pretreatment did not change type II collagen fibril formation. In conclusion, alk. pretreatment of sturgeon skin and notochord is an effective method to accelerate collagen extn. process of types I and II collagen without changing their biochem. properties. However, it decreases the yield of both collagens and specifically changes the fibril-forming ability of type I collagen.
- 35Furtado, M.; Chen, L.; Chen, Z.; Chen, A.; Cui, W. Development of Fish Collagen in Tissue Regeneration and Drug Delivery. Eng. Regen. 2022, 3, 217– 231, DOI: 10.1016/j.engreg.2022.05.002There is no corresponding record for this reference.
- 36Yoshimura, K.; Terashima, M.; Hozan, D.; Shirai, K. Preparation and Dynamic Viscoelasticity Characterization of Alkali- Solubilized Collagen from Shark Skin. J. Agric. Food Chem. 2000, 48, 685– 690, DOI: 10.1021/jf990389d36https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXhtlKjtrw%253D&md5=d138b61787c55e1d13b675ed31412e73Preparation and Dynamic Viscoelasticity Characterization of Alkali-Solubilized Collagen from Shark SkinYoshimura, Keiji; Terashima, Mariko; Hozan, Daiki; Shirai, KunioJournal of Agricultural and Food Chemistry (2000), 48 (3), 685-690CODEN: JAFCAU; ISSN:0021-8561. (American Chemical Society)Alkali-solubilized collagens, prepd. by alkali-acid extn. and alkali direct extn. (abbreviated AASC and ALSC, resp.), were characterized by dynamic viscoelastic measurement of collagen soln. (10 mg/mL). The optimum preparative conditions in terms of yield and polypeptide size are as follows: for the alkali-acid extn., a pretreatment with 0.5 or 1 M NaOH contg. 15% Na2SO4 within 5 days at 20 °C followed by the subsequent acid extn., and for the alk. direct extn., a treatment with 0.5 M NaOH contg. 10% NaCl at 4 °C for 20-30 days. A major portion of the polypeptide sizes of AASC and ALSC is composed of α chains (α1 and α2). Dynamic viscoelasticity of collagen soln. was measured as a function of temp. AASC showed a greater contribution of elastic behavior rather than viscous behavior. On the contrary, ALSC exhibits a stronger viscous behavior than elastic behavior.
- 37Bowes, J. H.; Kenten, R. H. The Effect of Alkalis on Collagen. Biochem. J. 1948, 43, 365– 372, DOI: 10.1042/bj043036537https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaH1MXitVSqtA%253D%253D&md5=163920312283531adbf4cb3b9cdf492dEffect of alkalies on collagenBowes, Joane H.; Kenten, R. H.Biochemical Journal (1948), 43 (), 365-72CODEN: BIJOAK; ISSN:0264-6021.Carefully prepd. and chemically unaltered collagen treated with alkali at pH 13 hydrolyzes off amide groups. This is the main reaction taking place. A small portion of guanidine groups is converted to ornithine and urea, and even a smaller portion to citrulline and NH3. Probably some peptide links of proline and hydroxyproline are broken. About 5% of the material becomes solubilized. Following alkali treatment (Ca(OH)2 + NaOH) the collagen swells more than before.
- 38Hattori, S.; Adachi, E.; Ebihara, T.; Shirai, T.; Someki, I.; Irie, S. Alkali-Treated the Ligand Collagen Activity Retained the the Adhesion Triple via Conformation Integrin for The Method of Extracting Collagen from Connective Tissue with a Solution of Sodium Hydroxide and Monomethylamine Was Developed about 30 Years Ago (. J. Biochem. 1999, 125, 676– 684, DOI: 10.1093/oxfordjournals.jbchem.a02233638https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXjsVyrtL4%253D&md5=9126263c0e61c88df79b4574d6bba37fAlkali-treated collagen retained the triple helical conformation and the ligand activity for the cell adhesion via α2β1 integrinHattori, Shunji; Adachi, Eijiro; Ebihara, Tetsuya; Shirai, Tomoko; Someki, Iori; Irie, ShinkichiJournal of Biochemistry (1999), 125 (4), 676-684CODEN: JOBIAO; ISSN:0021-924X. (Japanese Biochemical Society)Alk. treatment is a good method for extg. collagen with high recovery even from an aged animal specimen. However, the properties of collagen treated under alk. conditions have not been well established yet. By the treatment with a soln. of 3% sodium hydroxide and 1.9% monomethylamine, the isoelec. point of type I collagen was lowered from 9.3 to 4.8 because of the conversions of Asn and Gln to Asp and Glu. With the acidification of the pI, the denaturation temp. of the collagen was decreased from 42 to 35°C after 20 day treatment, but the collagen-specific triple helical conformation was maintained. Human keratinocytes and fibroblasts adhered to the alkali-treated collagen via the collagen receptor integrin α2β1. This indicates that the alkali-treated collagen maintained its property as a biol. adherent mol. Unlike acid-sol. collagen, alkali-treated collagen lost the ability to form fibrils at neutral pH under physiol. conditions. This ability was lost even after 4 h of alk. treatment, when the denaturation temp. of the collagen did not change. On the other hand, the alkali-treated collagen formed a fibrous ppt. with a uniform diam. of 50-70 nm under acidic conditions at 30°C.
- 39Yan, M.; Li, B.; Zhao, X.; Ren, G.; Zhuang, Y.; Hou, H.; Zhang, X.; Chen, L.; Fan, Y. Characterization of Acid-Soluble Collagen from the Skin of Walleye Pollock (Theragra Chalcogramma). Food Chem. 2008, 107, 1581– 1586, DOI: 10.1016/j.foodchem.2007.10.027There is no corresponding record for this reference.
- 40Nurubhasha, R.; Kumar, N. S. S.; Thirumalasetti, S. K.; Simhachalam, G.; Dirisala, V. R. Extraction and Characterization of Collagen from the Skin of Pterygoplichthys Pardalis and Its Potential Application in Food Industries. Food Sci. Biotechnol. 2019, 28, 1811– 1817, DOI: 10.1007/s10068-019-00601-z40https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtVCjtbbJ&md5=3e76fd31f97de51e50bf705eafa0d3ecExtraction and characterization of collagen from the skin of Pterygoplichthys pardalis and its potential application in food industriesNurubhasha, Ramesh; Sampath Kumar, N. S.; Thirumalasetti, Satish K.; Simhachalam, G.; Dirisala, Vijaya R.Food Science and Biotechnology (2019), 28 (6), 1811-1817CODEN: FSBOBR; ISSN:1226-7708. (Korean Society of Food Science and Technology)The primary objective of this study was to ext. collagen from underutilized fish species owing to its cost effective nature and also its ability to address the demand of type I collagen arising from food and pharmaceutical industries. Acid and pepsin sol. collagen (ppASC and ppPSC) were extd. from the skin of sucker catfish (Pterygoplichthys pardalis) with a yield of 19.6 and 23.8% on wet wt. basis resp. The same were characterized and confirmed as type I collagen by SDS-PAGE, FTIR and UV-Vis spectroscopy, amino acid anal., and Zeta potential. Taking into consideration the application of collagen in food industry, a food product was developed by incorporating with fresh cheese. This fortification was found to be acceptable and had not altered the taste, odor and other sensory properties of the product.
- 41Kamalvand, M.; Biazar, E.; Daliri-Joupari, M.; Montazer, F.; Rezaei-Tavirani, M.; Heidari-Keshel, S. Design of a Decellularized Fish Skin as a Biological Scaffold for Skin Tissue Regeneration. Tissue Cell 2021, 71, 101509 DOI: 10.1016/j.tice.2021.10150941https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXmsFGntr0%253D&md5=b30751ec998dce878cb954d080001676Design of a decellularized fish skin as a biological scaffold for skin tissue regenerationKamalvand, Mahshad; Biazar, Esmaeil; Daliri-Joupari, Morteza; Montazer, Fatemeh; Rezaei-Tavirani, Mostafa; Heidari-Keshel, SaeedTissue & Cell (2021), 71 (), 101509CODEN: TICEBI; ISSN:0040-8166. (Elsevier Ltd.)The use of decellularized natural skin as an extracellular matrix (ECM) may be a great candidate to regenerate damaged tissues. In this study, decellularized scaffolds from fish skin were designed by different techniques (phys., chem., and enzymic methods) and investigated by analyses such as Differential Scanning Calorimetry (DSC), SEM (SEM), Tensile strength, Degradability, Histol. studies, Toxicity test, and Detn. of DNA content. Results showed that the best sample is related to the decel lularized skin by hypertonic & hypotonic technique and Triton X100 solns. Structural and mech. results were demonstrated that samples have similar properties to human skin to regenerate it. The cytotoxicity results showed that decellularized skin by hypertonic & hypotonic method and Triton soln. is non-toxic with minimal amt. of genetic materials. Cellular results with epithelial cells indicated good adhesion on decellularized matrix, so it can be a suitable candidate for skin tissue regeneration.
- 42Lu, H. D.; Soranno, D. E.; Rodell, C. B.; Kim, I. L.; Burdick, J. A. Secondary Photocrosslinking of Injectable Shear-Thinning Dock-and-Lock Hydrogels. Adv. Healthcare Mater. 2013, 2, 1028– 1036, DOI: 10.1002/adhm.20120034342https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhtVOkt73F&md5=9d3f88bde9acfcd51e95a83666d1419eSecondary photocrosslinking of injectable shear-thinning dock-and-lock hydrogelsLu, Hoang D.; Soranno, Danielle E.; Rodell, Christopher B.; Kim, Iris L.; Burdick, Jason A.Advanced Healthcare Materials (2013), 2 (7), 1028-1036CODEN: AHMDBJ; ISSN:2192-2640. (Wiley-VCH Verlag GmbH & Co. KGaA)Shear-thinning hydrogels are useful in numerous applications, including as injectable carriers that act as scaffolds to support cell and drug therapies. Here, we describe the engineering of a self-assembling Dock-and-Lock (DnL) system that forms injectable shear-thinning hydrogels using mol. recognition interactions that also possess photo-triggerable secondary crosslinks. These DnL hydrogels are fabricated from peptide-modified hyaluronic acid (HA) and polypeptide precursors, can self-heal immediately after shear induced flow, are cytocompatible, and can be stabilized through light-initiated radical polymn. of methacrylate functional groups to tune gel mechanics and erosion kinetics. Secondary crosslinked hydrogels retain self-adhesive properties and exhibit cooperative phys. and chem. crosslinks with moduli as high as ∼10 times larger than moduli of gels based on phys. crosslinking alone. The extent of reaction and change in properties are dependent on whether the methacrylate is incorporated either at the terminus of the peptide or directly to the HA backbone. Addnl., the gel erosion can be monitored through an incorporated fluorophore and phys.-chem. gels remain intact in soln. over months, whereas phys. gels that are not covalently crosslinked erode completely within days. Mesenchymal stem cells exhibit increased viability when cultured in phys.- chem. gels, compared with those cultured in gels based on phys. crosslinks alone. The phys. properties of these DnL gels may be addnl. tuned by adjusting component compns., which allows DnL gels with a wide range of phys. properties to be constructed for use.
- 43Guo, S.; DiPietro, L. A. Critical Review in Oral Biology & Medicine: Factors Affecting Wound Healing. J. Dent. Res. 2010, 89, 219– 229, DOI: 10.1177/002203450935912543https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXjtFOnsLc%253D&md5=a42f3a6d9a5a275ced98dd32f6f7870fFactors affecting wound healingGuo, S.; Di Pietro, L. A.Journal of Dental Research (2010), 89 (3), 219-229CODEN: JDREAF; ISSN:0022-0345. (Sage Publications)A review. Wound healing, as a normal biol. process in the human body, is achieved through four precisely and highly programmed phases: hemostasis, inflammation, proliferation, and remodeling. For a wound to heal successfully, all four phases must occur in the proper sequence and time frame. Many factors can interfere with one or more phases of this process, thus causing improper or impaired wound healing. This article reviews the recent literature on the most significant factors that affect cutaneous wound healing and the potential cellular and/or mol. mechanisms involved. The factors discussed include oxygenation, infection, age and sex hormones, stress, diabetes, obesity, medications, alcoholism, smoking, and nutrition. A better understanding of the influence of these factors on repair may lead to therapeutics that improve wound healing and resolve impaired wounds.
- 44Kara, A.; Distler, T.; Polley, C.; Schneidereit, D.; Seitz, H.; Friedrich, O.; Tihminlioglu, F.; Boccaccini, A. R. 3D Printed Gelatin/Decellularized Bone Composite Scaffolds for Bone Tissue Engineering: Fabrication, Characterization and Cytocompatibility Study. Mater. Today Bio 2022, 15, 100309 DOI: 10.1016/j.mtbio.2022.10030944https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitFOmtbrM&md5=1ae6a6215b3ba1e06cf99b5f149c17733D printed gelatin/decellularized bone composite scaffolds for bone tissue engineering: Fabrication, characterization and cytocompatibility studyKara, Aylin; Distler, Thomas; Polley, Christian; Schneidereit, Dominik; Seitz, Hermann; Friedrich, Oliver; Tihminlioglu, Funda; Boccaccini, Aldo R.Materials Today Bio (2022), 15 (), 100309CODEN: MTBAC2; ISSN:2590-0064. (Elsevier Ltd.)Three-dimensional (3D) printing technol. enables the design of personalized scaffolds with tunable pore size and compn. Combining decellularization and 3D printing techniques provides the opportunity to fabricate scaffolds with high potential to mimic native tissue. The aim of this study is to produce novel decellularized bone extracellular matrix (dbECM)-reinforced composite-scaffold that can be used as a biomaterial for bone tissue engineering. Decellularized bone particles (dbPTs, ∼100 μm diam.) were obtained from rabbit femur and used as a reinforcement agent by mixing with gelatin (GEL) in different concns. 3D scaffolds were fabricated by using an extrusion-based bioprinter and crosslinking with microbial transglutaminase (mTG) enzyme, followed by freeze-drying to obtain porous structures. Fabricated 3D scaffolds were characterized morphol., mech., and chem. Furthermore, MC3T3-E1 mouse pre-osteoblast cells were seeded on the dbPTs reinforced GEL scaffolds (GEL/dbPTs) and cultured for 21 days to assess cytocompatibility and cell attachment. We demonstrate the 3D-printability of dbPTs-reinforced GEL hydrogels and the achievement of homogenous distribution of the dbPTs in the whole scaffold structure, as well as bioactivity and cytocompatibility of GEL/dbPTs scaffolds. It was shown that Young's modulus and degrdn. rate of scaffolds were enhanced with increasing dbPTs content. Multiphoton microscopy imaging displayed the interaction of cells with dbPTs, indicating attachment and proliferation of cells around the particles as well as into the GEL-particle hydrogels. Our results demonstrate that GEL/dbPTs hydrogel formulations have potential for bone tissue engineering.
- 45Shirazi, R.; Shirazi-Adl, A.; Hurtig, M. Role of Cartilage Collagen Fibrils Networks in Knee Joint Biomechanics under Compression. J. Biomech. 2008, 41, 3340– 3348, DOI: 10.1016/j.jbiomech.2008.09.03345https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BD1cjmvVGgsw%253D%253D&md5=fc8b4334d9b0ccd8f9954a0c761b51c8Role of cartilage collagen fibrils networks in knee joint biomechanics under compressionShirazi R; Shirazi-Adl A; Hurtig MJournal of biomechanics (2008), 41 (16), 3340-8 ISSN:0021-9290.Collagen fibrils networks in knee cartilage and menisci change in content and structure from a region to another. While resisting tension, they influence global joint response as well as local strains particularly at short-term periods. To investigate the role of fibrils networks in knee joint mechanics and in particular cartilage response, a novel model of the knee joint is developed that incorporates the cartilage and meniscus fibrils networks as well as depth-dependent properties in cartilage. The joint response under up to 2000N compression is investigated for conditions simulating the absence in cartilage of deep fibrils normal to subchondral bone or superficial fibrils parallel to surface as well as localized split of cartilage at subchondral junction or localized damage to superficial fibrils at loaded areas. Deep vertical fibrils network in cartilage play a crucial role in stiffening (by 10%) global response and protecting cartilage by reducing large strains (from maximum of 102% to 38%), in particular at subchondral junction. Superficial horizontal fibrils protect the tissue mainly from excessive strains at superficial layers (from 27% to 8%). Local cartilage split at base disrupts the normal function of vertical fibrils at the affected areas resulting in higher strains. Deep fibrils, and to a lesser extent superficial fibrils, play dominant mechanical roles in cartilage response under transient compression. Any treatment modality attempting to repair or regenerate cartilage defects involving partial or full thickness osteochondral grafts should account for the crucial role of collagen fibrils networks and the demanding mechanical environment of the tissue.
- 46Ying, H.; Zhou, J.; Wang, M.; Su, D.; Ma, Q.; Lv, G.; Chen, J. In Situ Formed Collagen-Hyaluronic Acid Hydrogel as Biomimetic Dressing for Promoting Spontaneous Wound Healing. Mater. Sci. Eng. C 2019, 101, 487– 498, DOI: 10.1016/j.msec.2019.03.09346https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXntFens7c%253D&md5=5c646f9422649322d527fd0d953f4c61In situ formed collagen-hyaluronic acid hydrogel as biomimetic dressing for promoting spontaneous wound healingYing, Huiyan; Zhou, Juan; Wang, Mingyu; Su, Dandan; Ma, Qiaoqiao; Lv, Guozhong; Chen, JinghuaMaterials Science & Engineering, C: Materials for Biological Applications (2019), 101 (), 487-498CODEN: MSCEEE; ISSN:0928-4931. (Elsevier B.V.)Wound dressing is distinctly important to wound healing, because it can not only protect wound from external disturbance, but also provide an ideal environment for wound closure. However, most of wound dressings need addnl. active ingredients to assist the repair process. In order to develop new dressings that can present spontaneous healing activity, herein, an injectable hydrogel consisted of collagen I and hyaluronic acid has been designed to mimic extracellular matrix for vascular cells growing and wound closure. The prepn. of hydrogel (COL-HA) was realized through in situ coupling of phenol moieties of collagen I-hydroxybenzoic acid (COL-P) and hyaluronic-acid-tyramine (HA-Tyr) through horseradish peroxidase (HRP). The phys. structure and properties were characterized, and the biol. performances were analyzed. COL-HA hydrogel presented porous structure that contributed to the exchange of gas, medium and nutrition. Human microvascular endothelial cells (HMEC) and fibroblasts (COS-7) cultured within this hydrogel showed significant proliferation behaviors. More importantly, a certain level of vascular endothelial growth factor (VEGF) was obsd. in HMEC cultured hydrogel, which led to the possibility of vascular regeneration. For the full-thickness wound, the healing ratio and validity of wound treated with COL-HA hydrogel were higher than com. drug and individual COL-P hydrogel, HA-Tyr hydrogel groups, since collagen and hyaluronic acid made joint efforts to improve wound repair.
- 47Uhl, F. E.; Zhang, F.; Pouliot, R. A.; Uriarte, J. J.; Rolandsson Enes, S.; Han, X.; Ouyang, Y.; Xia, K.; Westergren-Thorsson, G.; Malmström, A.; Hallgren, O.; Linhardt, R. J.; Weiss, D. J. Functional Role of Glycosaminoglycans in Decellularized Lung Extracellular Matrix. Acta Biomater. 2020, 102, 231– 246, DOI: 10.1016/j.actbio.2019.11.02947https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXitlehtL%252FO&md5=9b4dff340393b04d139016fd37ddaba6Functional role of glycosaminoglycans in decellularized lung extracellular matrixUhl, Franziska E.; Zhang, Fuming; Pouliot, Robert A.; Uriarte, Juan J.; Rolandsson Enes, Sara; Han, Xiaorui; Ouyang, Yilan; Xia, Ke; Westergren-Thorsson, Gunilla; Malmstrom, Anders; Hallgren, Oskar; Linhardt, Robert J.; Weiss, Daniel J.Acta Biomaterialia (2020), 102 (), 231-246CODEN: ABCICB; ISSN:1742-7061. (Elsevier Ltd.)Despite progress in use of decellularized lung scaffolds in ex vivo lung bioengineering schemes, including use of gels and other materials derived from the scaffolds, the detailed compn. and functional role of extracellular matrix (ECM) proteoglycans (PGs) and their glycosaminoglycan (GAG) chains remaining in decellularized lungs, is poorly understood. Using a commonly utilized detergent-based decellularization approach in human autopsy lungs resulted in disproportionate losses of GAGs with depletion of chondroitin sulfate/dermatan sulfate (CS/DS) > heparan sulfate (HS) > hyaluronic acid (HA). Specific changes in disaccharide compn. of remaining GAGs were obsd. with disproportionate loss of NS and NS2S for HS groups and of 4S for CS/DS groups. No significant influence of smoking history, sex, time to autopsy, or age was obsd. in native vs. decellularized lungs. Notably, surface plasmon resonance demonstrated that GAGs remaining in decellularized lungs were unable to bind key matrix-assocd. growth factors FGF2, HGF, and TGFβ1. Growth of lung epithelial, pulmonary vascular, and stromal cells cultured on the surface of or embedded within gels derived from decellularized human lungs was differentially and combinatorially enhanced by replenishing specific GAGs and FGF2, HGF, and TGFβ1. In summary, lung decellularization results in loss and/or dysfunction of specific GAGs or side chains significantly affecting matrix-assocd. growth factor binding and lung cell metab. GAG and matrix-assocd. growth factor replenishment thus needs to be incorporated into schemes for investigations utilizing gels and other materials produced from decellularized human lungs. In the current studies, we demonstrate that glycosaminoglycans (GAGs) are significantly depleted during decellularization and those that remain are dysfunctional and unable to bind matrix-assocd. growth factors crit. for cell growth and differentiation. Systematically repleting GAGs and matrix-assocd. growth factors to gels derived from decellularized human lung significantly and differentially affects cell growth. These studies highlight the importance of considering GAGs in decellularized lungs and their derivs.
- 48Linhardt, R. J.; Toida, T. Linhardt.ReviewGAG.121130.Pdf. Acc. Chem. Res. 2004, 37, 431– 438, DOI: 10.1021/ar030138x48https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXjvVShtbk%253D&md5=2211c39d916084933178e816466cedeeRole of glycosaminoglycans in cellular communicationLinhardt, Robert J.; Toida, ToshihikoAccounts of Chemical Research (2004), 37 (7), 431-438CODEN: ACHRE4; ISSN:0001-4842. (American Chemical Society)A review. Glycosaminoglycans are of crit. importance in intercellular communication in organisms. This ubiquitous class of linear polyanions interacts with a wide variety of proteins, including growth factors and chemokines, which regulate important physiol. processes. The presence of glycosaminoglycans on cell membranes and in the extracellular matrix also has resulted in their exploitation by infectious pathogens to gain access and entry into animal cells. Here, the authors have examd. the structural and phys. characteristics of these mols. responsible for their interaction with proteins important in cell-cell communication.
- 49West, J. D.; West, J. D.; Stamm, C. E.; Brown, H. A.; Justice, S. L.; Morano, K. A. Enhanced Toxicity of the Protein Cross-Linkers Divinyl Sulfone and Diethyl Acetylenedicarboxylate in Comparison to Related Monofunctional Electrophiles. Chem. Res. Toxicol. 2011, 24, 1457– 1459, DOI: 10.1021/tx200302w49https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXpvVygs74%253D&md5=b87c2368fb55d2f4d7de6f0c96f853fbEnhanced Toxicity of the Protein Cross-Linkers Divinyl Sulfone and Diethyl Acetylenedicarboxylate in Comparison to Related Monofunctional ElectrophilesWest, James D.; Stamm, Chelsea E.; Brown, Haley A.; Justice, Samantha L.; Morano, Kevin A.Chemical Research in Toxicology (2011), 24 (9), 1457-1459CODEN: CRTOEC; ISSN:0893-228X. (American Chemical Society)Previously, the authors detd. that di-Et acetylenedicarboxylate (DAD), a protein cross-linker, was significantly more toxic than analogous monofunctional electrophiles. The authors hypothesized that other protein cross-linkers enhance toxicity similarly. In agreement with this hypothesis, the bifunctional electrophile divinyl sulfone (DVSF) was 6-fold more toxic than Et vinyl sulfone (EVSF) in colorectal carcinoma cells and greater than 10-fold more toxic in Saccharomyces cerevisiae. DVSF and DAD caused oligomerization of yeast thioredoxin 2 (Trx2p) in vitro and promoted Trx2p crosslinking to other proteins in yeast at cytotoxic doses. These results suggest that protein crosslinking is considerably more detrimental to cellular homeostasis than simple alkylation.
- 50Fang, Y.; Han, Y.; Wang, S.; Chen, J.; Dai, K.; Xiong, Y.; Sun, B. Three-Dimensional Printing Bilayer Membranous Nanofiber Scaffold for Inhibiting Scar Hyperplasia of Skin. Biomater. Adv. 2022, 138, 212951 DOI: 10.1016/j.bioadv.2022.21295150https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitlOmu73P&md5=7bd7f1f553d62782a322eb27c6e56bc5Three-dimensional printing bilayer membranous nanofiber scaffold for inhibiting scar hyperplasia of skinFang, Yuan; Han, Yu; Wang, Shoubao; Chen, Jingting; Dai, Kerong; Xiong, Yao; Sun, BinbinBiomaterials Advances (2022), 138 (), 212951CODEN: BAIDBT; ISSN:2772-9508. (Elsevier B.V.)Hypertrophic scar (HS) is the manifestation of pathol. wound healing, which affects the beauty of patients, and even affects the normal phys. functions of patients. We aimed to develop a 3D printing layer membranous nanofiber scaffold similar to skin structure. Among them, poly (lactic-co-glycolic acid) copolymer (PLGA) nanofibers were used as the "epidermis" layer above, and a decellular dermis matrix (dECM) nanofiber scaffold was used as the "dermis" layer below. In vitro, exptl. results showed that PLGA and dECM nanofiber scaffolds had good biocompatibility. In vivo expts. showed that BLM nanofiber scaffolds could inhibit collagen fiber deposition and angiogenesis, to inhibit the formation of hypertrophic scars. This study shows a simple and effective method for preventing and inhibiting the formation of hypertrophic scars.
- 51Zhang, T.; Xu, H.; Zhang, Y.; Zhang, S.; Yang, X.; Wei, Y.; Huang, D.; Lian, X. Fabrication and Characterization of Double-Layer Asymmetric Dressing through Electrostatic Spinning and 3D Printing for Skin Wound Repair. Mater. Des. 2022, 218, 110711 DOI: 10.1016/j.matdes.2022.11071151https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38Xhtlams7rK&md5=8cdc5b7b68a3235097081a0f1a0b6395Fabrication and characterization of double-layer asymmetric dressing through electrostatic spinning and 3D printing for skin wound repairZhang, Ting; Xu, Hao; Zhang, Yonggang; Zhang, Siruo; Yang, Xia; Wei, Yan; Huang, Di; Lian, XiaojieMaterials & Design (2022), 218 (), 110711CODEN: MADSD2; ISSN:0264-1275. (Elsevier Ltd.)Ideal wound dressings provide optimal microenvironment for the reconstruction of damaged tissue. In this work, we fabricated a bilayer asym. dressing to mimic gradient structure of epidermis and dermis of skin by combining electrostatic spinning and 3D printing method with properties including surface hydrophilic and hydrophobic, porosity, mech. as well as antibacterial properties. The outer layer was prepd. by optimized PCL/PLA (PP) via electrostatic spinning to mimick epidermis with water repellency and against bacterial penetration, which has a tensile modulus of 19.69 ± 0.66 MPa. While, the inner layer was 3D printed by optimized sodium alginate/polyvinyl alc./chitosan quaternary ammonium salt (SPH). The tensile modulus of SPH with a porosity of 70-90% is 0.82 ± 0.01 MPa, and the water content can be achieved above 85%. The antibacterial efficacy of inner layer was tested against Staphylococcus aureus indicating forming inhibition zone with a diam. of 1.61 ± 0.35 cm. In addn., Cell Counting Kit-8 and Live/Dead assay was used to test the viability of human dermal fibroblasts (HFBS), which showed that PP/SPH with 6% PVA had not significant cytotoxic effects. The double-layer asym. dressing meets the requirements of skin mech. properties and provides an effective repair strategy for clin. skin trauma.
- 52Randall, M. J.; Jüngel, A.; Rimann, M.; Wuertz-Kozak, K. Advances in the Biofabrication of 3D Skin in Vitro: Healthy and Pathological Models. Front. Bioeng. Biotechnol. 2018, 6, 154 DOI: 10.3389/fbioe.2018.0015452https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BB3crgtFOksg%253D%253D&md5=63c5c7c795fc0b45203ba69b660e754cAdvances in the Biofabrication of 3D Skin in vitro: Healthy and Pathological ModelsRandall Matthew J; Wuertz-Kozak Karin; Jungel Astrid; Rimann Markus; Rimann Markus; Wuertz-Kozak Karin; Wuertz-Kozak KarinFrontiers in bioengineering and biotechnology (2018), 6 (), 154 ISSN:2296-4185.The relevance for in vitro three-dimensional (3D) tissue culture of skin has been present for almost a century. From using skin biopsies in organ culture, to vascularized organotypic full-thickness reconstructed human skin equivalents, in vitro tissue regeneration of 3D skin has reached a golden era. However, the reconstruction of 3D skin still has room to grow and develop. The need for reproducible methodology, physiological structures and tissue architecture, and perfusable vasculature are only recently becoming a reality, though the addition of more complex structures such as glands and tactile corpuscles require advanced technologies. In this review, we will discuss the current methodology for biofabrication of 3D skin models and highlight the advantages and disadvantages of the existing systems as well as emphasize how new techniques can aid in the production of a truly physiologically relevant skin construct for preclinical innovation.
- 53Kim, S.; Lee, H.; Kim, J. A.; Park, T. H. Prevention of Collagen Hydrogel Contraction Using Polydopamine-Coating and Alginate Outer Shell Increases Cell Contractile Force. Biomater. Adv. 2022, 136, 212780 DOI: 10.1016/j.bioadv.2022.21278053https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XitlOmurvP&md5=2d788e46a6d51bde1e6116e1f8b80d85Prevention of collagen hydrogel contraction using polydopamine-coating and alginate outer shell increases cell contractile forceKim, Seulha; Lee, Haein; Kim, Jeong Ah.; Park, Tai HyunBiomaterials Advances (2022), 136 (), 212780CODEN: BAIDBT; ISSN:2772-9508. (Elsevier B.V.)Collagen is the most abundant protein in the extracellular matrix of mammals and has a great effect on various cell behaviors including adhesion, differentiation, and migration. However, it is difficult to utilize collagen gel as a phys. scaffold in vitro because of its severe contraction. Decrease in the overall hydrogel vol. induces changes in cell distribution, and mass transfer within the gel. Uncontrolled mech. and physiol. factors in the fibrous matrix result in uncontrolled cell behaviors in the surrounding cells. In this study, two strategies were used to minimize the contraction of collagen gel. A disk-shaped frame made of polydopamine-coated polydimethylsiloxane (PDMS) prevented horizontal contraction at the edge of the hydrogel. The sequentially cross-linked collagen gel with alginate outer shell (CA-shell) structure inhibited the vertical gel contraction. The combined method synergistically prevented the hydrogel from shrinkage in long-term 3D cell culture. We obsd. the shift in balance of differentiation from adipogenesis to osteogenesis in mesenchymal stem cells under the environment where gel contraction was prevented, and confirmed that this phenomenon is closely assocd. with the mechanotransduction based on Yes-assocd. protein (YAP) localization. Development of this contraction inhibition platform made it possible to investigate the influence of regulation of cellular microenvironments. The phys. properties of the hydrogel fabricated in this study were similar to that of pure collagen gel but completely changed the cell behavior within the gel by inhibition of gel contraction. The platform can be used to broaden our understanding of the fundamental mechanism underlying cell-matrix interactions and reproduce extracellular matrix in vivo.
- 54Bacakova, M.; Pajorova, J.; Broz, A.; Hadraba, D.; Lopot, F.; Zavadakova, A.; Vistejnova, L.; Kostic, I.; Jencova, V.; Bacakova, L. Erratum to a Two-Layer Skin Construct Consisting of a Collagen Hydrogel Reinforced by a Fibrin-Coated Polylactide Nanofibrous Membrane (Int J Nanomedicine, (2019), 14, (5033–5050)). Int. J. Nanomed. 2019, 14, 7215– 7216, DOI: 10.2147/IJN.S228110There is no corresponding record for this reference.
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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.3c01642.
UV–vis spectra of solubilized fish-dECM before or after dialysis (Figure S1) and the water swelling ratio of 3D-printed models with 5 and 10% DVS (Figure S2) (PDF)
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