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Antiviral Polymer Brushes by Visible-Light-Induced, Oxygen-Tolerant Covalent Surface Coating

  • Andriy R. Kuzmyn
    Andriy R. Kuzmyn
    Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands
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    Orcidhttps://orcid.org/0000-0002-1571-2911
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  • Lucas W. Teunissen
    Lucas W. Teunissen
    Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands
    More by Lucas W. Teunissen
    Orcidhttps://orcid.org/0000-0003-0018-1211
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  • Michiel V. Kroese
    Michiel V. Kroese
    Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
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  • , 
  • Jet Kant
    Jet Kant
    Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
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  • , 
  • Sandra Venema
    Sandra Venema
    Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
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  • , and 
  • Han Zuilhof*
    Han Zuilhof
    Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands
    School of Pharmaceutical Sciences and Technology, Tianjin University, 92 Weijin Road, Tianjin 300072, People’s Republic of China
    Department of Chemical and Materials Engineering, Faculty of Engineering, King Abdulaziz University, 21589 Jeddah, Saudi Arabia
    *Email: [email protected]
    More by Han Zuilhof
    Orcidhttps://orcid.org/0000-0001-5773-8506
Cite this: ACS Omega 2022, 7, 43, 38371–38379
Publication Date (Web):October 20, 2022
https://doi.org/10.1021/acsomega.2c03214

Copyright © 2022 The Authors. Published by American Chemical Society. This publication is licensed under

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Abstract

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This work presents a novel route for creating metal-free antiviral coatings based on polymer brushes synthesized by surface-initiated photoinduced electron transfer-reversible addition–fragmentation chain transfer (SI-PET-RAFT) polymerization, applying eosin Y as a photocatalyst, water as a solvent, and visible light as a driving force. The polymer brushes were synthesized using N-[3-(decyldimethyl)-aminopropyl] methacrylamide bromide and carboxybetaine methacrylamide monomers. The chemical composition, thickness, roughness, and wettability of the resulting polymer brush coatings were characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), water contact angle measurements, and ellipsometry. The antiviral properties of coatings were investigated by exposure to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and avian influenza viruses, with further measurement of residual viable viral particles. The best performance was obtained with Cu surfaces, with a ca. 20-fold reduction of SARS-Cov-2 and a 50-fold reduction in avian influenza. On the polymer brush-modified surfaces, the number of viable virus particles decreased by about 5–6 times faster for avian flu and about 2–3 times faster for SARS-CoV-2, all compared to unmodified silicon surfaces. Interestingly, no significant differences were obtained between quaternary ammonium brushes and zwitterionic brushes.

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Introduction

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Viruses and virus-related diseases are one of the main reasons for human mortality. Recently, the world was engulfed by a pandemic induced by a novel human coronavirus 2 (severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)). The SARS-CoV-2 virus particles are roughly spherical with radii of 80–100 nm. (1,2) The exterior of the virus particles exhibits lipid-embedded, protein-based spikes of 10–20 nm, (2) which mediate the receptor binding and membrane fusion between the virus and host cell. SARS-CoV-2, in particular, showed a relatively high stability on different types of surfaces. Viable viruses were detected on plastic and stainless steel up to 72 h after virus exposure. (3) As a result, surface transmission is one of the possible ways of coronavirus transmittal. (4) Thus, innovative and effective approaches for inactivation and limitation of the spread of viruses via surfaces are paramount.
Antiviral properties are present in different types of surfaces, both of natural and artificial origin. Many plants have been demonstrated to contain antiviral compounds that can directly destroy viruses upon contact. (5) Heavy metals such as copper (Cu), gold (Au), and silver (Ag) have been used for centuries to combat the effects of viral infections (albeit, of course, only recognized as such in the last century). (6) For example, Ag nanoparticles have─by creating Ag+ ions─been shown to possess direct antiviral properties against a wide range of viruses, in particular, human immunodeficiency virus, hepatitis B virus, herpes simplex virus, respiratory syncytial virus, and monkeypox virus. (7) Cu-based surfaces have been shown to be highly effective against different classes of viruses, including enveloped and nonenveloped, single- and double-stranded and DNA and RNA viruses. (8,9) In some of these studies, the antiviral activity of such surfaces was demonstrated against SARS, Middle East respiratory syndrome (MERS), and SARS-CoV-2. (3,10) The virucidal activity of copper, similarly to silver, relies on the release of ionic Cu species (CuI or/and CuII) and, in some reports, generation of oxidative oxygen species, (8,10) and these positive results prompted us to include Cu surfaces in this study.
A different metal-free approach to creating virucidal surfaces invokes the use of antiviral polymers. (11) Polymers based on polyethyleneimine (PEI) (12,13) and polyphenol ether, (14) sialic acids, (15) and carbohydrates (16) have been reported to inactivate viruses. One of the approaches in creating antiviral coatings is the immobilization of polyethylenimine (PEI) derivatives containing long-chained hydrophobic quaternary ammonium compounds on surfaces. (13,14) Upon contact, these functional groups may disrupt the virus membrane structure rendering the virus inactive. N,N-Dodecyl,methyl-polyethylenimine (DMPEI) was found to be active against both wild-type and drug-resistant strains of influenza viruses by disrupting their lipid envelopes, resulting in visibly damaged viral particles. (17,18) That lipid envelope, known as the lipid bilayer, is the “scaffold” of the virus particle onto which functional proteins are bound, and within which the RNA is protected. Thus, disruption in the structure of the lipid layer compromises the structure of the virus. Consequently, it is a viable target in the inactivation of the virus.
Polymer brushes are high-density systems that are covalently attached to surfaces. (19,20) Coatings based on polymer brushes are extensively used to create antifouling, bioactive, and antibacterial surfaces. (21−37) The high density of the polymer chains in the coating on the surfaces gives unique properties for the final layer. (19,38,39) The most commonly used method for constructing polymer brushes is surface-initiated atom transfer radical polymerization (SI-ATRP), due to the high control and possibility to initiate growth from a self-assembled monolayer. (20,26) However, SI-ATRP requires the use of heavy metals, typically copper, as catalysts. Moreover, the need for rigorous deoxygenation to achieve polymerization makes this approach difficult to scale and experimentally challenging to apply. The ease of coating was further increased, and the possibility to surface patterning was introduced using techniques such as single-electron transfer living radical polymerization (SET-LRP) (34) and light-triggered living radical polymerization (LT-LRP), (22) but those methods still used heavy-metal catalysts. Thus, controlled polymerization approaches based on thermally initiated reversible addition-fragmentation chain transfer (RAFT) (40,41) polymerization and its surface-initiated (SI-RAFT) (24) analogue were introduced, which allow polymerizations without the need for heavy metals, although they still require an oxygen-free environment.
Recently, a new RAFT-based technique was introduced: photoinduced electron transfer-reversible addition-fragmentation chain transfer (PET-RAFT). (42−45) We and others further developed this method, labeled surface-initiated PET-RAFT, or SI-PET-RAFT, into an efficient and extremely mild (visible-light-driven, oxygen-tolerant, in water) way to covalently coat surfaces and applied this to the construction of polymer brushes. (21,45−47) This polymerization technique can proceed in an aqueous solution containing a suitably chosen dye, here: eosin Y (EY), as a photocatalyst and allows for the visible-light-induced patterned growth of complex polymer brushes in a controlled manner in the presence of oxygen in the water. A mechanism and the living nature of PET-RAFT polymerizations were previously postulated by Xu et al. (48) They proposed that the reaction proceeds according to a reductive quenching cycle of eosin Y in which triethanolamine acts as a sacrificial electron donor to reduce oxygen in the polymerization system. The reduction of oxygen allows the polymerization to proceed in an oxygen-containing environment, which is the main advantage of this technique in contrast to SI-ATRP and SI-RAFT. We, therefore, envisioned SI-PET-RAFT to be a potentially viable route for the construction of quaternary ammonium-based antiviral coatings.
Finally, since surface roughness has also been reported as a likely factor in antiviral activity, (11) while we aimed to study the specific effects of the chemical functionalities involved, only smooth surfaces (i.e., nonstructured and atomic force microscopy (AFM)-measured roughness ≤3.5 nm) were used.
This paper investigates the antiviral properties of copper surfaces and polymer brushes containing quaternary ammonium moieties as constructed with the SI-PET-RAFT technique. A new methacrylic derivative of N,N-dodecyl,methyl-polyethylenimine, N-[3-decyldimethyl-aminopropyl] methacrylamide bromide (A10) was synthesized for such polymer brushes. We envisioned that polymer brushes based on A10 monomers would be able to render viruses inactive by penetrating their membranes (Scheme 1). In addition, carboxybetaine methacrylamide (CBMA)-based brushes previously showed antibacterial and antifouling properties. (21,49,50) To the best of our knowledge, however, no antiviral studies have been reported for these brushes, and these A10- and CBMA-coated surfaces were now tested in parallel for their antiviral properties. The chemical composition, thickness, roughness, and wettability of the resulting polymer brush coatings were characterized extensively by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), water contact angle measurements, and ellipsometry. The antiviral properties of coatings were investigated by exposure to SARS-CoV-2 and avian influenza and measured in terms of the amount of viable virus after a specific time. The economic and humanitarian damages of the annually returning influenza virus pandemics are also well analyzed and studied, (51) yet the structure of avian flu virus is rather different from that of SARS-CoV-2, motivating why this virus type was chosen as the second candidate for this model study to better display the scope of our findings.

Scheme 1

Scheme 1. (Top) General Scheme of Polymer Brush-Based Antiviral Surfaces; (Bottom) Structure of Monomers A10 and CBMA Used to Construct Quaternary Ammonium and Zwitterionic Polymer Brushes
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Experimental Section

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Materials

All chemical reagents were used without further purification unless otherwise specified. 4-Cyano-4-(phenylcarbonothioylthio)pentanoic acid N-succinimidyl ester (RAFT-NHS), N-[3-(dimethylamino)propyl]methacrylamide, 1-bromodecane, (3-aminopropyl)triethoxysilane (APTES), triethanolamine (TEOA), eosin Y (EY), triethylamine (TEA), ethanol (EtOH, 99.9%), acetone (99.5%), dry tetrahydrofuran (THF, 99.9%), dry diethyl ether (99.9%), and acetonitrile (99.9%) were purchased from Sigma-Aldrich. Carboxybetaine methacrylamide (CBMA) was synthesized according to a previously described procedure. (31,52,53) Silicon substrates were acquired from Siltronix. Deionized water was produced with a Milli-Q integral 3 system (Millipore, Molsheim, France (Milli-Q water)).

Synthesis of N-[3-(Alkyldimethyl)-aminopropyl]methacrylamide Bromide Monomer (A10)

Monomer A10 was synthesized via a quaternization reaction of N-[3-(dimethylamino)propyl]methacrylamide and 1-bromodecane, according to a slight modification of a previously reported procedure. (54) In a round-bottom flask with a septum, 10 mL of acetonitrile was added to a mixture of N-[3-(dimethylamino)propyl]methacrylamide (0.03 mol, 5.10 g, 4.79 mL), 1-bromodecane (0.03 mol, 6.64 g), and a small amount of an inhibitor (hydroquinone). The flask was sealed with a septum under argon and heated at 50 °C under magnetic stirring for 24 h. Upon addition of 250 mL of dry ethyl ether to the reaction mixture, N-[3-(decyldimethyl)-aminopropyl]methacrylamide bromide (A10) was isolated as white crystals and dried (7.00 g, yield 60%). The monomer was characterized by 1H NMR and electrospray ionization (ESI).
N-[3-(Decyldimethyl)-aminopropyl]methacrylamide bromide (A10) (Figure S1): 1H NMR (dimethyl sulfoxide (DMSO), 400 MHz, δ in ppm): 0.86 (t, 3H, CH3 (22)), 1.26 (m, 14H, CH2 (15, 16, 17, 18, 19, 20, 21)), 1.62 (m, 2H, CH2 (14)), 1.87 (m, 5H, CH2 (6, 9)), 3.01 (s, 6H, N+–CH3 (11, 12)), 3.25 (m, 6H, CH2 (5, 7, 13)), 5.35 (s, 1H, HC═CCH3 CH2(1″)), 5.70 (s, 1H, HC═C(1′)), 8.10 (t, 1H, NH) (Figure S1). Mass ESI: measured: 311.3048. theoretical (M+): 311.3062.

Mass Spectrometry (MS)

MS data were recorded on an Exactive high-resolution MS instrument (Thermo Scientific) equipped with an electrospray ionization (ESI) probe. The MS was calibrated daily using ProteoMass LTQ/FT-hybrid ESI Pos. Mode Cal. Mix and Pierce ESI Neg. Ion Cal. solutions. Thermo Xcalibur Browser software version 2.2 was used for instrument control, data acquisition, and data processing. The ESI mass spectra were obtained over the range of m/z 100–750. The exact mass of compound A10 was determined with a precision of 3 ppm.

Light Source

Light-emitting diodes (LEDs) with a maximum intensity at 410 nm (Intelligent LED Solutions product number: ILH-XO01-S410-SC211-WIR200) were used. The input current was set at 700 mA, corresponding to a total radiometric power of 2.9 W, according to manufacturer specifications.

Formation of RAFT Agent-Functionalized Monolayers

The monolayers were created according to previously published procedures. (21,47)

SI-PET-RAFT Synthesis of Polymer Brushes

The poly(CBMA) brushes were synthesized by a previously published technique. (21,47) The poly(A10) brushes were prepared according to a previously reported protocol with slight modifications. (21,47) A stock solution with photocatalyst was prepared to contain EY (25 mg, 39 μmol) and TEOA (160 mg, 1.6 mmol) in 10 mL of Milli-Q water. The monomer A10 (0.31 mmol, 122 mg) was dissolved in Milli-Q water (1 mL), and subsequently, 10 μL of the stock solution was added. The mixture was vortexed and added to the vials containing surfaces with an immobilized RAFT agent. Immediately after this, the polymerization was conducted by irradiating the vials with visible light from the LED light source for controlled periods of time. The thickness of the polymerization solution on top of the surfaces was 2 mm. In these experiments, the light source was placed 3–4 cm from the substrates, to prevent substantial heating of the samples with the light. Afterward, a copious amount of EtOH was added to the samples to remove unbound polymers from the surface, and further cleaning was obtained by subsequent rinsing with EtOH and Milli-Q water. Finally, the coated samples were blown dry under a stream of Ar.
In addition, the reaction was conducted on a complete 100 mm silicon wafer to provide the necessary amount of surfaces for the antiviral tests. The polymerization solution was scaled up from 1 to 30 mL, and for this purpose, a custom-made reactor 200 × 200 × 2.0 mm3 was designed and used (see setup in Figure S2).

X-ray Photoelectron Spectroscopy (XPS)

XPS measurements were performed using a JPS-9200 photoelectron spectrometer (JEOL Ltd., Japan). All of the samples were prepared and stored under ambient conditions prior to analysis using a focused monochromated Al Kα X-ray source (spot size of 300 μm) radiation at 12 kV and 20 mA, with 10 eV as analyzer pass energy. XPS wide-scan and narrow-scan spectra were obtained under ultrahigh-vacuum (UHV) conditions (base pressure 3 × 10–7 Pa). All narrow-range spectra were corrected with a linear background before fitting. The spectra were fitted with symmetrical Gaussian/Lorentzian (GL(30)) line shapes using CasaXPS. All spectra were referenced to the C 1s peak attributed to C–C and C–H atoms at 285.0 eV. (55)

Static Water Contact Angle (SWCA) Measurements

The wettability of the modified surfaces was determined by automated static water contact angle measurements with the use of a Kruss DSA 100 goniometer. The volume of a drop of demineralized water was 3 μL. Contact angles from sessile drops measured by the tangent method were estimated using a standard error propagation technique involving partial derivatives.

Spectroscopic Ellipsometry

Polymerization kinetics were followed by measuring the dry thickness of the brushes using an Accurion Nanofilm_ep4 Imaging Ellipsometer. The ellipsometric data were acquired in air at room temperature (RT) using light in the wavelength range of λ = 400.6–761.3 nm at an angle of incidence of 50°. The data were fitted with EP4 software using a multilayer model. The model consisted of a silicon (Si) bottom layer covered with a thin SiO2 layer of 0.3 nm. Then, the polymer brush layers were described using a Cauchy model with parameters A = 1.526 and B = 1221 for poly(CBMA) and A = 1.540 and B = 334 for poly(A10).

Atomic Force Microscopy (AFM)

AFM surface topography images were acquired by an Asylum Research MFP-3D Origin AFM (Oxford Instruments, U.K.). The instrument was operated in tapping mode and equipped with a silicon cantilever (AC240TS-R3, k = 1.3 N·m–1) with a nominal tip radius of ∼7 nm. Gwyddion (56) and Mountains 8 (Digital Surf, France) software was used to process and analyze the AFM topography images.

Antiviral Activity

The following low pathogenic avian influenza strain was used: H5N2 A/Swan/Neth/11016034/11. To culture avian influenza and access the amount of viable virus, Madin–Darby Canine Kidney (MDCK) cells were maintained in complete medium: Dulbecco’s modified Eagle’s medium, high glucose, GlutaMAX, pyruvate + 5.0% fetal bovine serum (FBS) + 0.01% penicillin–streptomycin (10,000 U·mL–1; to prevent bacterial and fungal contamination). When MDCK cells were inoculated with avian influenza, the complete medium was substituted with Dulbecco’s modified Eagle’s medium, high glucose, GlutaMAX, pyruvate + 1.5% albumin, bovine, reagent grade + 0.5 μg·mL–1 trypsin from bovine pancreas.
To evaluate the antiviral properties of the polymer brushes against avian influenza, 50 μL of virus stock (dilution 1:1000) was dispersed onto the coated antiviral surfaces. After incubation periods at room temperature (RT) of 1.5, 3, 7, 24, and 72 h, possible viable virus was washed off using 500 μL of complete medium. For each time point, at least three separate surfaces were used. For the determination of viral titers, eight 10-fold serial dilutions were used to inoculate MDCK cells (described in Bergervoet et al. (57)) with the addition of 0.5 μg trypsin·mL–1. After incubation at 37 °C for 4 days, plates were stained with an immunoperoxidase monolayer assay and scored for positive (stained) wells (57) with the substitution of paraformaldehyde by 4% formaldehyde. Titers were calculated using the Spearman–Kärber algorithm and were expressed as TCID50·mL–1. (58,59)

SARS-CoV-2

The SARS-CoV-2 strain used was described by Gerhards et al. (60) Vero-E6 cells were maintained in complete medium: minimum essential medium, no glutamine + 5% fetal bovine serum (FBS, qualified, Australia) + 1% l-glutamine (200 mM) + 1% minimum essential medium non-essential amino acids solution (100×) + 1% antibiotic–antimycotic solution (penicillin and streptomycin, to prevent bacterial and fungal contamination).
To access the antiviral properties of the polymer brushes against SARS-CoV-2, 50 μL of virus stock (undiluted) was dispersed onto the coated antiviral surfaces. After incubation periods at RT of 1.5, 3, 7, 24, and 72 h, a possible viable virus was washed off using 500 μL of complete medium. For each time point, at least three separate surfaces were used. For the determination of viral titers, six 5-fold serial dilutions were used to inoculate Vero-E6 cells. (60) After incubation at 37 °C for 4 days, plates were stained with an immunoperoxidase monolayer assay and scored for positive (stained) wells. (60) Titers were calculated using the Spearman–Kärber algorithm and were expressed as TCID50·mL–1. (58,59) All SARS-CoV-2 experiments were performed at the BSL3 facilities in Lelystad.

Results and Discussion

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The two monomers selected for the creation of potential polymer brush-based antiviral coatings were N-[3-decyldimethyl-aminopropyl]methacrylamide bromide (A10) and carboxybetaine methacrylate (CBMA). The A10 monomer was synthesized via the quaternization reaction of N-[3-(dimethylamino)propyl]methacrylamide and 1-bromodecane according to a slight modification of a previously reported procedure (see the Experimental Section for details). (54) The antiviral polymer brush-based coatings were created in four steps starting from bare silicon surfaces according to a modified previously published protocol (Scheme 2). (21,47) The silicon surfaces were plasma cleaned for 5 min, and subsequently, (3-aminopropyl)triethoxysilane (APTES) was immobilized. The RAFT-agent monolayer was prepared by reacting the (3-aminopropyl)triethoxysilane (APTES) monolayer, immobilized on the oxidized silicon surface, with 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid N-succinimidyl ester (RAFT-NHS). In this process, 29 ± 4% of the surface-bound amine sites react to hold a RAFT agent moiety. (21) The polymer brushes were then grown employing SI-PET-RAFT in the presence of the corresponding monomer (A10 or CBMA), EY, and TEOA.

Scheme 2

Scheme 2. General Scheme for the Synthesis of Antiviral Polymer Brushes
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Synthesis of Antiviral Polymer Brush-Based Coatings

The thickness of brushes was determined by spectroscopic ellipsometry. The poly(A10) brush-based coatings achieved a thickness of 71 ± 2 nm after 1 h of polymerization, while the poly(CBMA) brush-based coating grew to 46 ± 4 nm after 5 h of polymerization. The AFM topography images of brush-coated surfaces showed homogeneous layers with a roughness of Rq = 3.50 ± 0.55 and 2.03 ± 0.49 nm for poly(A10) and poly(CBMA), respectively (Figures 1d,e and S5). The static water contact angle (SWCA) of coatings for poly(A10) was determined to be 88 ± 4° and for poly(CBMA) 37 ± 2°. The relatively higher SWCA of poly(CBMA) brushes compared to polymer brushes obtained by ATRP (36) may be related not only to the high molecular weight of the resulting polymer but also to other factors like polymer confirmation, surface tension, and surface roughness. (61) In addition, the SWCA values are well aligned with previously reported values for CBMA obtained by nitroxide-mediated free radical polymerization-based coatings. (62) The chemical composition of the resulting polymer brushes was confirmed by XPS (Figure 1a). The wide spectra of poly(CBMA) brushes showed three main peaks for O 1s (531 eV), C 1s (285 eV), and N 1s (400 eV) in a ratio of 3.0:13.5:1.7, in reasonable agreement with the theoretical ratio of poly(CBMA) brushes, 3:12:2. The poly(A10) brushes were characterized using XPS as well, and the wide spectra showed four main peaks, for O 1s (531 eV), C 1s (285 eV), N 1s (400 eV), and Br 3s (255 eV) (secondary peaks: Br 3p (181 eV) and Br 3d (69 eV)) in a ratio of 1.0:23.8:2.2:0.8, while the theoretical ratio was 1:19:2:1. The relatively higher amount of carbon in both polymer brush structures is attributed to atmospheric contamination.

Figure 1

Figure 1. Physicochemical properties of polymer brush-coated surfaces. XPS wide spectra of poly(CBMA)- and poly(A10)-coated surfaces (a). XPS N 1s narrow scan (b) and XPS C 1s narrow scan (c) of poly(CBMA)- and poly(A10)-coated surfaces. Typical AFM topography images of poly(A10)- (d) and poly(CBMA)-coated surfaces (e).

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The XPS C 1s narrow-scan spectrum for poly(A10) shows three main peaks, which we assign as [C–C/H] (285.0 eV)/[C–N] (286.2 eV)/[HN–C═O] (287.8 eV) moieties in a ratio of 13.0:5.1:1.0 (theoretical ratio is: 13:5:1), and for poly(CBMA), these peaks appear in a ratio of 2.7:2.4:1.0 (theoretical ratio: 2.5:2.5:1.0) (Figure 1c). The narrow XPS spectra for the poly(A10) brushes were simulated applying core orbital energy levels as obtained by density functional theory (DFT), (63,64) and the simulated spectra (Figure S3) agreed well with experimental data, supporting our assignment. The narrow N 1s spectra of poly(CBMA) and poly(A10) brushes show two peaks that correspond to [N+] (403 eV) and [NH] (400 eV) in ratios of 0.63:1.00 and 0.83:1.00, respectively (Figure 1b). The relatively lower than theoretically expected value [for both monomers 1:1] can be related to the degradation of [N+] moieties under heat, in vacuo conditions, and exposure to X-ray beams during the XPS measurement, as noted for poly(CBMA) coatings by van Andel et al. (50)

Copper-Coated Surfaces

A custom-produced silicon wafer was used that was homogeneously coated with a copper layer (thickness Cu layer = 200 nm; AFM-determined roughness Rq= 1.74 ± 0.11 nm, SWCA 94 ± 3°; see Figure S4).

Control Surfaces

Neutral unmodified plasma-cleaned silicon surfaces, with a measured roughness of Rq= 0.68 ± 0.10 nm, were used as a negative control (Figure S4b). The wide-scan XPS spectra of these silicon surfaces showed the predominance of silicon oxide (Figure S4a).

Antiviral Studies

We exposed our surfaces to SARS-CoV-2 and avian influenza. In a typical experiment, a 50 μL drop of virus stock containing SARS-CoV-2 and avian influenza was deposited on the different types of surfaces. These solutions contained for SARS-CoV-2 ca. 69,000 viable viral particles·mL–1 and for avian influenza ca. 54,800 viable viral particles·mL–1. The drop was left on the surface for different periods of time, ranging from 1.5 to 72 h (Figure 2 and Tables S1 and S2). Subsequently, surfaces were washed, and the exact number of viable viral particles was determined by TCID50·mL–1 titration (see the Experimental Section for details).

Figure 2

Figure 2. Antiviral activity of copper-coated surfaces, and poly(CBMA) and poly(A10) brush surfaces against SARS-CoV-2 and avian influenza. Plasma-cleaned silicon surfaces were used as a negative control.

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First, the copper-coated surfaces showed the best antiviral activity toward both SARS-CoV-2 and avian influenza. Compared to our negative control (SiO2), copper inactivates SARS-CoV-2 at least 20 times faster, while for avian influenza, the number after 1.5 h is about 50 times smaller than on silicon oxide. These results indicate that, in cases where the toxicity of copper ions does not play a significant role, infusion of copper might be a viable manner to turn a surface antiviral. This high activity of copper is in line with related studies of SARS-CoV-2 and other viruses. (3,8−10) Likely, part of the virus-killing mechanism also affects cells, making copper both a good antiviral agent and toxic for humans. Therefore, metal-free solutions are also badly needed.
Second, the amount of SARS-CoV-2 viable virus on the poly(CBMA) and poly(A10) after 1.5 h of incubation was two to three times lower compared to the bare silicon surface. The polymer brush coatings incubated with avian influenza virus decreased the amount of viable virus in comparison to bare silicon surfaces even up to 5–6 times. After 3 h of incubation of viable SARS-CoV-2 and avian influenza virus on poly(CBMA) and poly(A10) surfaces decreased 4–5 times and 3–4 times correspondingly in relation to the amount of virus detected after 1.5 h of incubation. Furthermore, brush-based coatings inactivated most of the deposited virus on the surfaces within 7 h of incubation. Although difficult to compare quantitatively, this compares positively to SAR-CoV-2 surviving even up to 72 h on steel and plastic, (3) and these findings might imply that our “negative control” displays some antiviral activity as well. The poly(CBMA) and poly(A10) brushes thus overall show comparable antiviral activity toward SARS-CoV-2 and avian influenza. While not as effective as copper, the poly(CBMA) and poly(A10) coatings do not contain heavy metals. Moreover, poly(CBMA)-based coatings and polymers previously demonstrated good antifouling and biocompatible properties. (49,50,65) A probable antiviral mechanism for poly(A10) brushes, as previously reported for DMPEI compounds mechanics, (17,18) is based on penetration of the virus membrane and rendering it inactive (Scheme 3). Such a route would likely not apply for poly(CBMA), which does not have large aliphatic tails in its structure, yet shows a comparable antiviral activity. Previously, zwitterionic and cationic PEI derivatives (N-(15-carboxypentadecyl)-PEI HCl salt) containing long aliphatic chains in the comparable study showed similar antiviral activity. (13) It should be noted that CBMA is overall a much smaller moiety with more closely spaced charge positions and significantly higher hydrophilicity than previously investigated PEI zwitterionic derivatives. Thus, the pathway toward virus dismantling may be also related to the presence of quaternary ammonium cation in poly(CBMA) and poly(A10) brushes. The mechanism of antiviral activity of both brush coatings can also be absolutely different. While the positive charge and hydrophobic part play a key role in dismantling the virus for A10 brushes, the strong affinity to water and previously reported antifouling properties of the CBMA brush might explain the antiviral activity of those brushes. It should be noted that at this stage, the mechanism of antiviral activity of the brushes is unknown and may be related to many factors including the combination superstructure of the brush itself and positively charged moieties along the chain.

Scheme 3

Scheme 3. General Scheme of the Suggested Antiviral Mechanism of Lipid Bilayer-Disrupting Polymer Brushes (1–3); Monomer Chemical Structure and Monomer Schematic Representation Are Shown Below
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The effect of the porosity on the micron scale was previously reported as an important factor in virus survival on the surface. (66) In our study, both the synthesized surfaces and control surfaces are all basically flat (AFM topography displays roughnesses only in the range of a few nanometers; see Figures 1d,e and S3). As a result, differences in porosity or roughness likely play no significant role in determining the differences observed in our measurements.
Finally, the facile way to apply SI-PET-RAFT for creating polymer brush coatings, requiring only visible light to supply the driving force, might open up a range of applications where alternatives─from ventilating rooms to frequent cleaning─do not suffice. One can, for example, envision such coatings on common frequent-contact surfaces, such as plastic cards, touch screens, door handles, and walls. Here, the minimal toxicity of bioinert polymer brushes, in particular, poly(CBMA) brushes, would allow application in areas that would be unthinkable for toxic coatings such as copper. Eventually, such applications could also be applied to plastics such as PPE or rubbery materials to help diminish the risk of sexually transmitted infections or those transmittable by touch. Finally, it is likely that the combination of chemical and physical effects together (chemically well-chosen functional groups on a surface with optimal micro/nanostructuring, for example) will be needed for optimal results. Such work is currently ongoing in our laboratories.

Conclusions

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A new route was developed for creating antiviral coatings based on copper coating and on polymer brushes synthesized by surface-initiated photoinduced electron transfer-reversible addition-fragmentation chain transfer (SI-PET-RAFT). Copper-coated surfaces are very effective, speeding up the inactivation of the virus about 25–50 times compared to a silicon oxide negative control. Both quaternary ammonium polymer brushes and zwitterionic polymer brushes strongly, and more or less equally, sped up the process of inactivating viruses, with a reduction of factor 5–6 times for avian flu and a factor 2–3 times for SARS-CoV-2. In both cases, nearly all virus activity reached down to detection levels after about 7 h. Since SI-PET-RAFT is scalable, oxygen-tolerant, driven by visible light, can take place in water, and is applicable to almost any surface, we envision the significant potential of our approach in the further study and development of antiviral surfaces, likely in combination with the optimization of surface structuring.

Supporting Information

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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsomega.2c03214.

  • 1H NMR spectrum of N-[3-(decyldimethyl)-aminopropyl]methacrylamide bromide (A10); scale-up photoreactor scheme; simulated C 1s XPS spectra; wide-scan XPS spectra of the silicon and copper surfaces used in antiviral tests and AFM topography; viable viral particles of SARS-CoV-2 on different surfaces at different periods of time; viable viral particles of avian influenza on different surfaces at different periods of time (PDF)

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Author Information

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  • Corresponding Author
    • Han Zuilhof - Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands;  School of Pharmaceutical Sciences and Technology, Tianjin University, 92 Weijin Road, Tianjin 300072, People’s Republic of China;  Department of Chemical and Materials Engineering, Faculty of Engineering, King Abdulaziz University, 21589 Jeddah, Saudi Arabia;  Orcidhttps://orcid.org/0000-0001-5773-8506;  Email: [email protected]
  • Authors
    • Andriy R. Kuzmyn - Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands;  Orcidhttps://orcid.org/0000-0002-1571-2911
    • Lucas W. Teunissen - Laboratory of Organic Chemistry, Wageningen University, Stippeneng 4, 6708 WE Wageningen, The Netherlands;  Orcidhttps://orcid.org/0000-0003-0018-1211
    • Michiel V. Kroese - Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
    • Jet Kant - Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
    • Sandra Venema - Wageningen Bioveterinary Research, Houtribweg 39, 8221 RA Lelystad, The Netherlands
  • Author Contributions

    The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

  • Funding

    This research was carried out under project number T20001 in the framework of the Research Program of the Materials innovation institute (M2i) (www.m2i.nl) supported by the Dutch government.

  • Notes
    The authors declare no competing financial interest.

Acknowledgments

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Dr. Maarten M. J. Smulders is acknowledged for insightful discussions.

References

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    Protocols for the synthesis of the microbicidal polycation N,N-dodecyl,methyl-polyethylenimine and coating (painting) of glass slides with this polycation's butanol soln. are described. Subsequently detailed are the procedures for validating that the resultant coated slides are essentially 100% lethal to the human bacterial pathogens, Staphylococcus aureus and Escherichia coli, as well as to two common strains of influenza virus. The time required to prep. and apply the cationic polymer and to test its microbicidal efficiency is conservatively estd. to be <4 wk.
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  19. 19
    Michalek, L.; Barner, L.; Barner-Kowollik, C. Polymer on Top: Current Limits and Future Perspectives of Quantitatively Evaluating Surface Grafting. Adv. Mater. 2018, 30, 1706321  DOI: 10.1002/adma.201706321
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    Zoppe, J. O.; Ataman, N. C.; Mocny, P.; Wang, J.; Moraes, J.; Klok, H.-A. Surface-Initiated Controlled Radical Polymerization: State-of-the-Art, Opportunities, and Challenges in Surface and Interface Engineering with Polymer Brushes. Chem. Rev. 2017, 117, 1105– 1318,  DOI: 10.1021/acs.chemrev.6b00314
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    20
    Surface-Initiated Controlled Radical Polymerization: State-of-the-Art, Opportunities, and Challenges in Surface and Interface Engineering with Polymer Brushes
    Zoppe, Justin O.; Ataman, Nariye Cavusoglu; Mocny, Piotr; Wang, Jian; Moraes, John; Klok, Harm-Anton
    Chemical Reviews (Washington, DC, United States) (2017), 117 (3), 1105-1318CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)
    A review. The generation of polymer brushes by surface-initiated controlled radical polymn. (SI-CRP) techniques has become a powerful approach to tailor the chem. and phys. properties of interfaces and has given rise to great advances in surface and interface engineering. Polymer brushes are defined as thin polymer films in which the individual polymer chains are tethered by one chain end to a solid interface. Significant advances have been made over the past years in the field of polymer brushes. This includes novel developments in SI-CRP, as well as the emergence of novel applications such as catalysis, electronics, nanomaterial synthesis and biosensing. Addnl., polymer brushes prepd. via SI-CRP have been utilized to modify the surface of novel substrates such as natural fibers, polymer nanofibers, mesoporous materials, graphene, viruses and protein nanoparticles. The last years have also seen exciting advances in the chem. and phys. characterization of polymer brushes, as well as an ever increasing set of computational and simulation tools that allow understanding and predictions of these surface-grafted polymer architectures. The aim of this contribution is to provide a comprehensive review that critically assesses recent advances in the field and highlights the opportunities and challenges for future work.
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  21. 21
    Kuzmyn, A. R.; Nguyen, A. T.; Teunissen, L. W.; Zuilhof, H.; Baggerman, J. Antifouling Polymer Brushes via Oxygen-Tolerant Surface-Initiated PET-RAFT. Langmuir 2020, 36, 4439– 4446,  DOI: 10.1021/acs.langmuir.9b03536
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    21
    Antifouling Polymer Brushes via Oxygen-Tolerant Surface-Initiated PET-RAFT
    Kuzmyn, Andriy R.; Nguyen, Ai T.; Teunissen, Lucas W.; Zuilhof, Han; Baggerman, Jacob
    Langmuir (2020), 36 (16), 4439-4446CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    This work presents a new method for the synthesis of antifouling polymer brushes using surface-initiated photoinduced electron transfer-reversible addn.-fragmentation chain-transfer polymn. with eosin Y and triethanolamine as catalysts. This method proceeds in an aq. environment under atm. conditions without any prior degassing and without the use of heavy metal catalysts. The versatility of the method is shown by using three chem. different monomers: oligo(ethylene glycol) methacrylate, N-(2-hydroxypropyl)methacrylamide, and carboxybetaine methacrylamide. In addn., the light-triggered nature of the polymn. allows the creation of complex three-dimensional structures. The compn. and topol. structuring of the brushes are confirmed by XPS and at. force microscopy. The kinetics of the polymns. are followed by measuring the layer thickness with ellipsometry. The polymer brushes demonstrate excellent antifouling properties when exposed to single-protein solns. and complex biol. matrixes such as dild. bovine serum. This method thus presents a new simple approach for the manufg. of antifouling coatings for biomedical and biotechnol. applications.
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  22. 22
    Kuzmyn, A. R.; Nguyen, A. T.; Zuilhof, H.; Baggerman, J. Bioactive Antifouling Surfaces by Visible-Light-Triggered Polymerization. Adv. Mater. Interfaces 2019, 6, 1900351  DOI: 10.1002/admi.201900351
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    Pop-Georgievski, O.; Rodriguez-Emmenegger, C.; Pereira, A. d. l. S.; Proks, V.; Brynda, E.; Rypáček, F. Biomimetic Non-fouling Surfaces: Extending the Concepts. J. Mater. Chem. B 2013, 1, 2859– 2867,  DOI: 10.1039/c3tb20346h
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    23
    Biomimetic non-fouling surfaces: extending the concepts
    Pop-Georgievski, Ognen; Rodriguez-Emmenegger, Cesar; de los Santos Pereira, Andres; Proks, Vladimir; Brynda, Eduard; Rypacek, Frantisek
    Journal of Materials Chemistry B: Materials for Biology and Medicine (2013), 1 (22), 2859-2867CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)
    In this study, we propose a substrate-independent biomimetic modification route for the creation of antifouling polymer brushes. This modification route consists of the formation/deposition of a biomimetic polydopamine anchor layer followed by a well-controlled surface-initiated atom transfer radical polymn. of antifouling polymer brushes initiated by 2-bromo-2-methylpropanoyl groups covalently attached to the hydroxyl and amine groups present in the anchor layer. In this way, we synthesized polymer brushes of methoxy- and hydroxy-capped oligoethylene glycol methacrylate, 2-hydroxyethyl methacrylate and carboxybetaine acrylamide. Spectroscopic ellipsometry (SE) indicated well-controlled polymn. kinetics of the brushes, thus the thickness of the ultra-thin films could be precisely tuned at a nanometer scale. The covalent structure and organization of the brushes grown from the polydopamine anchor layer were accessed by IR reflection-adsorption spectroscopy (IRRAS) while the change in hydrophilicity caused by the presence of the brush was detd. by dynamic water contact angle measurements. Surface plasmon resonance as well as ex situ IRRAS and SE measurements were applied to investigate the adsorption of model protein solns. and undiluted human blood plasma to the brushes. The biomimetic brushes completely suppressed the fouling from single protein solns. and reduced the fouling from plasma to less than 3% from the fouling measured on bare gold surfaces. The proposed modification procedure is non-destructive and does not require any chem. pre-activation or the presence of reactive groups on the substrate surface. Contrary to other antifouling modifications the coating can be performed on various classes of substrates and preserves its properties even in undiluted blood plasma. This work offers a promising technol. for the facile fabrication of different surface-based biotechnol. and biomedical devices able to perform tailor-made functions while resisting the fouling from the complex biol. media where they operate.
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  24. 24
    Zamfir, M.; Rodriguez-Emmenegger, C.; Bauer, S.; Barner, L.; Rosenhahn, A.; Barner-Kowollik, C. Controlled Growth of Protein Resistant PHEMA Brushes via S-RAFT Polymerization. J. Mater. Chem. B 2013, 1, 6027– 6034,  DOI: 10.1039/c3tb20880j
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    24
    Controlled growth of protein resistant PHEMA brushes via S-RAFT polymerization
    Zamfir, Mirela; Rodriguez-Emmenegger, Cesar; Bauer, Stella; Barner, Leonie; Rosenhahn, Axel; Barner-Kowollik, Christopher
    Journal of Materials Chemistry B: Materials for Biology and Medicine (2013), 1 (44), 6027-6034CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)
    The reversible addn.-fragmentation chain transfer polymn. of 2-hydroxyethyl methacrylate (HEMA) from surfaces (S-RAFT) using an R-group-attached chain transfer agent (CTA) is presented. The approach was exploited for the efficient prepn. of well-defined PHEMA brushes of up to 50 nm thickness in a controlled fashion without using any cytotoxic catalyst. The chem. compn., morphol. and wettability of the samples were assessed by XPS, at. force microscopy and water contact angle measurements, while the growth kinetics were studied by monitoring the dry thickness via spectroscopic ellipsometry. The mechanism and kinetics of the RAFT polymn. on the surface - in the presence of a sacrificial CTA and of solvent mixts. with different polarities - were investigated. A marked effect of the concn. of the sacrificial CTA on the kinetics was obsd. Importantly - and for the first time - the living PHEMA brushes were exploited as macroRAFT agents for chain extension, and thicknesses up to 70 nm were achieved. The prepd. PHEMA brushes were challenged with protein solns. demonstrating their resistance to fouling.
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  25. 25
    Lange, S. C.; van Andel, E.; Smulders, M. M. J.; Zuilhof, H. Efficient and Tunable Three-Dimensional Functionalization of Fully Zwitterionic Antifouling Surface Coatings. Langmuir 2016, 32, 10199– 10205,  DOI: 10.1021/acs.langmuir.6b02622
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    25
    Efficient and Tunable Three-Dimensional Functionalization of Fully Zwitterionic Antifouling Surface Coatings
    Lange, Stefanie C.; van Andel, Esther; Smulders, Maarten M. J.; Zuilhof, Han
    Langmuir (2016), 32 (40), 10199-10205CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    In order to enhance the sensitivity and selectivity of surface-based (bio)sensors, it is of crucial importance to diminish background signals that arise from the nonspecific binding of biomols., so-called biofouling. Zwitterionic polymer brushes have been shown to be excellent antifouling materials. However, for sensing purposes, antifouling does no suffice, but need to be combined with the possibility to efficiently modify the brush with recognition units. So far this has only been achieved at the expense of either antifouling properties or binding capacity. Herein the authors present a conceptually new approach by integrating both characteristics into a single, tailor-made monomer: a novel sulfobetaine-based zwitterionic monomer equipped with a clickable azide moiety. Copolymn. of this monomer with a well-established std. sulfobetaine monomer, results in highly antifouling surface coatings with a high, yet tunable amt. of clickable groups present throughout the entire brush. Subsequent functionalization of the azido-brushes, via e.g. widely used strain-promoted alkyne azide click reactions yields fully zwitterionic 3D-functionalized coatings with a recognition unit of choice that can be tailored for any specific application. Here the authors show a proof-of-principle with biotin-functionalized brushes on Si3N4 that combine excellent antifouling properties with specific avidin binding from a protein mixt. The signal-to-noise ratio is significantly improved over traditional chain end modification of sulfobetaine polymer brushes, even if the azide content is lowered to 1%, thus offering a viable approach for the development of significantly improved performance of biosensors on any surface.
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  26. 26
    Kuzmyn, A. R.; de los Santos Pereira, A.; Pop-Georgievski, O.; Bruns, M.; Brynda, E.; Rodriguez-Emmenegger, C. Exploiting End Group Functionalization for the Design of Antifouling Bioactive Brushes. Polym. Chem. 2014, 5, 4124– 4131,  DOI: 10.1039/C4PY00281D
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    26
    Exploiting end group functionalization for the design of antifouling bioactive brushes
    Kuzmyn, A. R.; de los Santos Pereira, A.; Pop-Georgievski, O.; Bruns, M.; Brynda, E.; Rodriguez-Emmenegger, C.
    Polymer Chemistry (2014), 5 (13), 4124-4131CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
    Biol. active surfaces are essential in many applications in the fields of biosensing, bioimplants, and tissue engineering. However, the introduction of bioactive motifs without impairment of their ability to resist non-specific interactions with biol. media remains a challenge. Herein, we present a straightforward, facile strategy for the creation of bioactive surfaces based on the end-group biofunctionalization of state-of-the-art polymer brushes via an ultra-fast Diels-Alder "click" reaction. Surface-initiated atom transfer radical polymn. is employed to grow antifouling polymers preserving the end groups. These groups are then further converted to a reactive cyclopentadienyl moiety and exploited for the immobilization of biomols. on the topmost layer of the brush. The minimal chem. modification of the antifouling polymer brush accounts for the full preservation of the fouling resistance of the surface even after biofunctionalization, which is crit. for the aforementioned applications.
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  27. 27
    Rodriguez Emmenegger, C.; Brynda, E.; Riedel, T.; Sedlakova, Z.; Houska, M.; Alles, A. B. Interaction of Blood Plasma with Antifouling Surfaces. Langmuir 2009, 25, 6328– 6333,  DOI: 10.1021/la900083s
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    27
    Interaction of blood plasma with antifouling surfaces
    Rodriguez Emmenegger C; Brynda E; Riedel T; Sedlakova Z; Houska M; Alles A Bologna
    Langmuir : the ACS journal of surfaces and colloids (2009), 25 (11), 6328-33 ISSN:0743-7463.
    Nonspecific adsorption of proteins is a crucial problem in the detection of analytes in complex biological media by affinity sensors operating with label-free detection. We modified the gold surface of surface plasmon resonance (SPR) sensors with three types of promising antifouling coatings: self-assembled monolayers (SAM)s of alkanethiolates terminated with diethylene glycol and carboxylic groups, poly(ethylene glycol) (PEG) grafted onto the SAMs, and zwitterionic polymer brushes of poly(carboxybetaine methacrylate), poly(sulfobetaine methacrylate), and poly(phosphorylcholine methacrylate). Using SPR, we compared the efficacy of the coatings to reduce nonspecific adsorption from human blood plasma and from single-protein solutions of human serum albumin, immunoglobulin G, fibrinogen, and lysozyme. There was no direct relationship between values of water contact angles and plasma deposition on the coated surfaces. A rather high plasma deposition on SAMs was decreased by grafting PEG chains. Fouling on PEG was observed only from plasma fractions containing proteins with molecular mass higher than 350 000 Da. The adsorption kinetics from plasma collected from different healthy donors differed. Poly(carboxybetaine methacrylate) completely prevented the deposition from plasma, but the other more hydrophilic zwitterionic polymers prevented single-protein adsorption but did not prevent plasma deposition. The results suggest that neither wettability nor adsorption of the main plasma proteins was the main indicator of deposition from blood plasma.
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    Williams, D. F. On the Nature of Biomaterials. Biomaterials 2009, 30, 5897– 5909,  DOI: 10.1016/j.biomaterials.2009.07.027
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    28
    On the nature of biomaterials
    Williams, David F.
    Biomaterials (2009), 30 (30), 5897-5909CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
    A review. The situations in which biomaterials are currently used are vastly different to those of just a decade ago. Although implantable medical devices are still immensely important, medical technologies now encompass a range of drug and gene delivery systems, tissue engineering and cell therapies, organ printing and cell patterning, nanotechnol. based imaging and diagnostic systems and microelectronic devices. These technologies still encompass metals, ceramics and synthetic polymers, but also biopolymers, self assembled systems, nanoparticles, carbon nanotubes and quantum dots. These changes imply that our original concepts of biomaterials and our expectations of their performance also have to change. This Leading Opinion Paper addresses these issues. It concludes that many substances which hitherto we may not have thought of as biomaterials should now be considered as such so that, alongside the traditional structural biomaterials, we have substances that have been engineered to perform functions within health care where their performance is directly controlled by interactions with tissues and tissue components. These include engineered tissues, cells, organs and even viruses. This essay develops the arguments for a radically different definition of a biomaterial.
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  29. 29
    Rodriguez-Emmenegger, C.; Avramenko, O. A.; Brynda, E.; Skvor, J.; Alles, A. B. Poly(HEMA) Brushes Emerging as a New Platform for Direct Detection of Food Pathogen in Milk Samples. Biosens. Bioelectron. 2011, 26, 4545– 4551,  DOI: 10.1016/j.bios.2011.05.021
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    29
    Poly(HEMA) brushes emerging as a new platform for direct detection of food pathogen in milk samples
    Rodriguez-Emmenegger, Cesar; Avramenko, Oxana A.; Brynda, Eduard; Skvor, J.; Alles, Aldo Bologna
    Biosensors & Bioelectronics (2011), 26 (11), 4545-4551CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
    Surface plasmon resonance (SPR) biosensors capable of in real time detection of Cronobacter at concns. down to 106 cells mL-1 in samples of consumer fresh-whole fat milk, powder whole-fat milk prepn., and powder infant formulation were developed for the first time. Antibodies against Cronobacter were covalently attached onto polymer brushes of poly(2-hydroxyethyl methacrylate) (poly(HEMA)) grafted from the SPR chip surface. The lowest detection limit, 104 cells mL-1, was achieved in phosphate buffered saline (pH 7.4) with sensors prepd. by covalent immobilization of the same antibodies onto a self assembled monolayer (SAM) of hexa(ethylene glycol) undecanethiol (EG6). However, when the EG6 based sensors were challenged with milk samples the non-specific response due to the deposition of non-targeted compds. from the milk samples was much higher than the specific response to Cronobacter hampering the detection in milk. Similar interfering fouling was obsd. on antifouling polymer brushes of hydroxy-capped oligoethylene glycol methacrylate and even a 10 times higher fouling was obsd. on the widely used SAM of mixed hydroxy- and carboxy-terminated alkanethiols. Only poly(HEMA) brushes totally suppressed the fouling from milk samples. The robust well-controlled surface initiated atom transfer radical polymn. of HEMA allowed the prepn. of highly dense brushes with a minimal thickness so that the capture of antigens by the antibodies immobilized on the brush layer could take place close to the gold SPR surface to provide a stronger optical response while the fouling was still suppressed. A min. thickness of 19 nm of poly(HEMA) brush layer was necessary to suppress completely non-specific sensor response to fouling from milk.
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  30. 30
    Liu, H.; Wang, S. Poly(N-isopropylacrylamide)-based thermo-responsive surfaces with controllable cell adhesion. Sci. China: Chem. 2014, 57, 552– 557,  DOI: 10.1007/s11426-013-5051-1
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    Poly(N-isopropylacrylamide)-based thermo-responsive surfaces with controllable cell adhesion
    Liu, Hong Liang; Wang, Shu Tao
    Science China: Chemistry (2014), 57 (4), 552-557CODEN: SCCCCS; ISSN:1869-1870. (Science China Press)
    A review. Poly(N-isopropylacrylamide) (PNIPAAm)-based thermo-responsive surfaces can switch their wettability (from wettable to non-wettable) and adhesion (from sticky to non-sticky) according to external temp. changes. These smart surfaces with switchable interfacial properties are playing increasingly important roles in a diverse range of biomedical applications; these controlling cell-adhesion behavior has shown great potential for tissue engineering and disease diagnostics. Herein we reviewed the recent progress of research on PNIPAAm-based thermo-responsive surfaces that can dynamically control cell adhesion behavior. The underlying response mechanisms and influencing factors for PNIPAAm-based surfaces to control cell adhesion are described first. Then, PNIPAAm-modified two-dimensional flat surfaces for cell-sheet engineering and PNIPAAm-modified three-dimensional nanostructured surfaces for diagnostics are summarized. We also provide a future perspective for the development of stimuli-responsive surfaces.
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  31. 31
    Rodriguez-Emmenegger, C.; Brynda, E.; Riedel, T.; Houska, M.; Šubr, V.; Alles, A. B.; Hasan, E.; Gautrot, J. E.; Huck, W. T. S. Polymer Brushes Showing Non-Fouling in Blood Plasma Challenge the Currently Accepted Design of Protein Resistant Surfaces. Macromol. Rapid Commun. 2011, 32, 952– 957,  DOI: 10.1002/marc.201100189
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    Polymer Brushes Showing Non-Fouling in Blood Plasma Challenge the Currently Accepted Design of Protein Resistant Surfaces
    Rodriguez-Emmenegger, Cesar; Brynda, Eduard; Riedel, Tomas; Houska, Milan; Subr, Vladimir; Alles, Aldo Bologna; Hasan, Erol; Gautrot, Julien E.; Huck, Wilhelm T. S.
    Macromolecular Rapid Communications (2011), 32 (13), 952-957CODEN: MRCOE3; ISSN:1022-1336. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Ultra-low-fouling poly[N-(2-hydroxypropyl) methacrylamide] (poly(HPMA)) brushes have been synthesized for the first time. Similar to the so far only ultra-low-fouling surface, poly(carboxybetaine acrylamide), the level of blood plasma fouling was below the detection limit of surface plasmon resonance (SPR, 0.03 ng/cm-2) despite being a hydrogen bond donor and displaying a moderate wettability, thus challenging the currently accepted views for the design of antifouling properties. The antifouling properties were preserved even after two years of storage. To demonstrate the potential of poly(HPMA) brushes for the prepn. of bioactive ultra-low fouling surfaces a label-free SPR immunosensor for detection of G Streptococcus was prepd.
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    Baggerman, J.; Smulders, M. M. J.; Zuilhof, H. Romantic Surfaces: A Systematic Overview of Stable, Biospecific, and Antifouling Zwitterionic Surfaces. Langmuir 2019, 35, 1072– 1084,  DOI: 10.1021/acs.langmuir.8b03360
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    Romantic Surfaces: A Systematic Overview of Stable, Biospecific, and Antifouling Zwitterionic Surfaces
    Baggerman, Jacob; Smulders, Maarten M. J.; Zuilhof, Han
    Langmuir (2019), 35 (5), 1072-1084CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    A review. This feature Article focuses on recent advances in the bioconjugation of surface-bound zwitterionic polymers for biospecific antifouling surfaces. Various approaches for the functionalization of antifouling zwitterionic polymers are systematically investigated, such as chain-end and side-chain functionalization. Side-chain functionalization methods can be further classified as those that are achieved through homopolymn. of custom-synthesized zwitterionic monomers equipped with reactive groups, or those that are achieved via synthesis of random or block copolymers combining different monomers with antifouling functionality and others with reactive groups. Several of the pros and cons of these approaches are outlined and discussed. Finally, some perspective and future directions of research are presented toward long-term stable, generically repelling surfaces that strongly and specifically adhere to a single component in a complex mixt.
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    Wischerhoff, E.; Badi, N.; Lutz, J.-F.; Laschewsky, A. Smart Bioactive Surfaces. Soft Matter 2010, 6, 705– 713,  DOI: 10.1039/B913594D
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    Smart bioactive surfaces
    Wischerhoff, Erik; Badi, Nezha; Lutz, Jean-Francois; Laschewsky, Andre
    Soft Matter (2010), 6 (4), 705-713CODEN: SMOABF; ISSN:1744-683X. (Royal Society of Chemistry)
    A review. The purpose of this highlight is to define the emerging field of bioactive surfaces. In recent years, various types of synthetic materials capable of "communicating" with biol. objects such as nucleic acids, proteins, polysaccharides, viruses, bacteria or living cells were described in the literature. This novel area of research certainly goes beyond the traditional field of smart materials and includes different types of sophisticated interactions with biol. entities, such as reversible adhesion, conformational control, biol.-triggered release and selective permeation. These novel materials may be 2D planar surfaces as well as colloidal objects or 3D scaffolds. Overall, they show great promise for numerous applications in biosciences and biotechnol. For instance, practical applications of bioactive surfaces in the fields of biosepn., cell engineering, biochips and stem-cell differentiation are briefly discussed herein.
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    Vorobii, M.; de los Santos Pereira, A.; Pop-Georgievski, O.; Kostina, N. Y.; Rodriguez-Emmenegger, C.; Percec, V. Synthesis of Non-fouling poly[N-(2-hydroxypropyl)methacrylamide] Brushes by Photoinduced SET-LRP. Polym. Chem. 2015, 6, 4210– 4220,  DOI: 10.1039/C5PY00506J
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    Synthesis of non-fouling poly[N-(2-hydroxypropyl)methacrylamide] brushes by photoinduced SET-LRP
    Vorobii, Mariia; de los Santos Pereira, Andres; Pop-Georgievski, Ognen; Kostina, Nina Yu.; Rodriguez-Emmenegger, Cesar; Percec, Virgil
    Polymer Chemistry (2015), 6 (23), 4210-4220CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
    Surface-initiated photoinduced single-electron transfer living radical polymn. (SET-LRP) was employed to assemble brushes of poly[N-(2-hydroxypropyl) methacrylamide] (poly(HPMA)) from Si surfaces. The linear increase in thickness of the poly(HPMA) brushes with time and the ability to prep. block copolymers indicate the living nature of this grafting-from process. Cu concns. ≥80 ppb were sufficient for this surface-initiated SET-LRP. Micropatterns of poly(HPMA) brushes on the Si surface were constructed for the first time by this method. Negligible fouling was obsd. after contact with undiluted blood plasma. This report provides the first example of nonfouling polymer brushes prepd. by SET-LRP of HPMA.
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    Jiang, S.; Cao, Z. Ultralow-Fouling, Functionalizable, and Hydrolyzable Zwitterionic Materials and Their Derivatives for Biological Applications. Adv. Mater. 2010, 22, 920– 932,  DOI: 10.1002/adma.200901407
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    Ultralow-Fouling, Functionalizable, and Hydrolyzable Zwitterionic Materials and Their Derivatives for Biological Applications
    Jiang, Shaoyi; Cao, Zhiqiang
    Advanced Materials (Weinheim, Germany) (2010), 22 (9), 920-932CODEN: ADVMEW; ISSN:0935-9648. (Wiley-VCH Verlag GmbH & Co. KGaA)
    A review. In recent years, zwitterionic materials such as poly(carboxybetaine) (pCB) and poly(sulfobetaine) (pSB) have been applied to a broad range of biomedical and engineering materials. Due to electrostatically induced hydration, surfaces coated with zwitterionic groups are highly resistant to nonspecific protein adsorption, bacterial adhesion, and biofilm formation. Among zwitterionic materials, pCB is unique due to its abundant functional groups for the convenient immobilization of biomols. pCB can also be prepd. in a hydrolyzable form as cationic pCB esters, which can kill bacteria or condense DNA. The hydrolysis of cationic pCB esters into nonfouling zwitterionic groups will lead to the release of killed microbes or the irreversible unpackaging of DNA. Furthermore, mixed-charge materials have been shown to be equiv. to zwitterionic materials in resisting nonspecific protein adsorption when they are uniformly mixed at the mol. scale.
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    Pereira, A. d. l. S.; Rodriguez-Emmenegger, C.; Surman, F.; Riedel, T.; Alles, A. B.; Brynda, E. Use of Pooled Blood Plasmas in the Assessment of Fouling Resistance. RSC Adv. 2014, 4, 2318– 2321,  DOI: 10.1039/C3RA43093F
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    Use of pooled blood plasmas in the assessment of fouling resistance
    Pereira, Andres de los Santos; Rodriguez-Emmenegger, Cesar; Surman, Frantisek; Riedel, Tomas; Alles, Aldo Bologna; Brynda, Eduard
    RSC Advances (2014), 4 (5), 2318-2321CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)
    The ability of a surface to resist fouling from blood plasma cannot be realistically estd. without measuring adsorption from real samples directly. Due to the variability of biol. samples, pooled blood plasma is normally used. We show that even when using pooled plasma, a comparison of antifouling surfaces strongly depends on its source.
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    Teunissen, L. W.; Kuzmyn, A. R.; Ruggeri, F. S.; Smulders, M. M. J.; Zuilhof, H. Thermoresponsive, Pyrrolidone-Based Antifouling Polymer Brushes. Adv. Mater. Interfaces 2022, 9, 2101717  DOI: 10.1002/admi.202101717
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    Thermoresponsive, pyrrolidone-based antifouling polymer brushes
    Teunissen, Lucas W.; Kuzmyn, Andriy R.; Ruggeri, Francesco S.; Smulders, Maarten M. J.; Zuilhof, Han
    Advanced Materials Interfaces (2022), 9 (6), 2101717CODEN: AMIDD2; ISSN:2196-7350. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Commonly, modification of surfaces with thermoresponsive polymers is performed using poly(N-isopropylacrylamide) (poly(NIPAM)). However, integration of poly(NIPAM) with a second polymer to obtain more complex copolymer structures has proven challenging due to inherently poorly controllable polymn. characteristics of acrylamides. In this study, N-(2-methacryloyloxyethyl)pyrrolidone (NMEP) is synthesized and polymd. under controlled conditions from silicon oxide substrates via surface-initiated atom transfer radical polymn. (SI-ATRP) to produce thermoresponsive poly(NMEP) brushes. The livingness of the brushes is demonstrated by reinitiation of poly(NMEP) brushes using oligo(ethylene glycol) Me ether methacrylate to obtain diblock copolymer brushes. Following extensive characterization, the reversible thermoresponsive behavior of these poly(NMEP) brushes is demonstrated using phase-controlled AFM topog. measurements in an aq. liq. environment. These measurements indicate that at 27°C the poly(NMEP) brushes are solvated and extend away from the surface. Finally, to investigate the potential applicability of poly(NMEP) brushes in biomedical devices, the antifouling properties of the coating are tested in aq. media contg. BSA, fibrinogen, or 10% dild. human serum using quartz crystal microbalance with dissipation monitoring (QCM-D). These measurements reveal very good antifouling properties, even when exposed to 10% dild. human serum.
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    van der Weg, K. J.; Ritsema van Eck, G. C.; de Beer, S. Polymer Brush Friction in Cylindrical Geometries. Lubricants 2019, 7, 84  DOI: 10.3390/lubricants7100084
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    Yu, Y.; Brió Pérez, M.; Cao, C.; de Beer, S. Switching (Bio-) adhesion and Friction in Liquid by Stimulus Responsive Polymer Coatings. Eur. Polym. J. 2021, 147, 110298  DOI: 10.1016/j.eurpolymj.2021.110298
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    Switching (bio-) adhesion and friction in liquid by stimulus responsive polymer coatings
    Yu, Yunlong; Brio Perez, Maria; Cao, Cong; de Beer, Sissi
    European Polymer Journal (2021), 147 (), 110298CODEN: EUPJAG; ISSN:0014-3057. (Elsevier Ltd.)
    A review. Controlling (bio-) adhesion and friction is essential in many bio-medical and industrial applications. Such control can be achieved using stimulus responsive polymers. Coatings composed of these polymers are able to adapt their physicochem. properties to changes in the surrounding environment, such as temp., light, pH, electronic and magnetic fields. This responsive behavior can be employed in the development of advanced technologies as for example underwater adhesives, self-adhering wound dressings, soft robotics and targeted drug delivery. In this review, we present an overview of the current status of research on controlling adhesion and friction in liq. environments using surface-bound stimulus responsive polymers. We focus in particular on polymer brushes and the role of specific interactions between the surfaces in the contact. These interactions can give rise to complex relations between swelling of the polymer film and its tribomech. properties. A careful characterization of the change in specific interactions upon varying the environment is, therefore, crit. for the design of effective smart coatings. We will finish this review with an overview of the open questions in the field and a discussion of the potential impact that the polymer-based smart adhesives and lubricants can have on industry and society.
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    Yeole, N. Thiocarbonylthio Compounds. Synlett 2010, 2010, 1572– 1573,  DOI: 10.1055/s-0029-1219938
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    Moad, G.; Rizzardo, E.; Thang, S. H. Radical addition–fragmentation chemistry in polymer synthesis. Polymer 2008, 49, 1079– 1131,  DOI: 10.1016/j.polymer.2007.11.020
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    Radical addition-fragmentation chemistry in polymer synthesis
    Moad, Graeme; Rizzardo, Ezio; Thang, San H.
    Polymer (2008), 49 (5), 1079-1131CODEN: POLMAG; ISSN:0032-3861. (Elsevier Ltd.)
    A review on the development of addn.-fragmentation chain transfer agents and related ring-opening monomers highlighting recent innovation in these areas. The major part of this review deals with reagents that give reversible addn.-fragmentation chain transfer (RAFT). These reagents include dithioesters, trithiocarbonates, dithiocarbamates and xanthates. The RAFT process is a versatile method for conferring living characteristics on radical polymns. providing unprecedented control over mol. wt., mol. wt. distribution, compn. and architecture. It is suitable for most monomers polymerizable by radical polymn. and is robust under a wide range of reaction conditions. It provides a route to functional polymers, cyclopolymers, gradient copolymers, block polymers and star polymers.
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    Niu, J.; Page, Z. A.; Dolinski, N. D.; Anastasaki, A.; Hsueh, A. T.; Soh, H. T.; Hawker, C. J. Rapid Visible Light-Mediated Controlled Aqueous Polymerization with In Situ Monitoring. ACS Macro Lett. 2017, 6, 1109– 1113,  DOI: 10.1021/acsmacrolett.7b00587
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    Rapid Visible Light-Mediated Controlled Aqueous Polymerization with In Situ Monitoring
    Niu, Jia; Page, Zachariah A.; Dolinski, Neil D.; Anastasaki, Athina; Hsueh, Andy T.; Soh, H. Tom; Hawker, Craig J.
    ACS Macro Letters (2017), 6 (10), 1109-1113CODEN: AMLCCD; ISSN:2161-1653. (American Chemical Society)
    We report a simple procedure for rapid, visible light-mediated, controlled radical polymn. in aq. solns. Based on the photoelectron transfer reversible addn.-fragmentation chain transfer (PET-RAFT) polymn., fast chain propagation at room temp. in water was achieved in the presence of reductant and without prior deoxygenation. A systematic study correlating irradn. intensity and polymn. kinetics, enabled by in situ NMR spectroscopy, provided optimized reaction conditions. The versatility of this procedure was demonstrated through a rapid triblock copolymer synthesis, and incorporation of water-labile activated esters for direct conjugation of hydrophilic small mols. and proteins. In addn., this technique boasts excellent temporal control and provides a wide range of macromol. materials with controlled mol. wts. and narrow mol. wt. distributions.
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    Lueckerath, T.; Strauch, T.; Koynov, K.; Barner-Kowollik, C.; Ng, D. Y. W.; Weil, T. DNA–Polymer Conjugates by Photoinduced RAFT Polymerization. Biomacromolecules 2019, 20, 212– 221,  DOI: 10.1021/acs.biomac.8b01328
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    DNA-Polymer Conjugates by Photoinduced RAFT Polymerization
    Lueckerath, Thorsten; Strauch, Tina; Koynov, Kaloian; Barner-Kowollik, Christopher; Ng, David Y. W.; Weil, Tanja
    Biomacromolecules (2019), 20 (1), 212-221CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)
    Conventional grafting-to approaches to DNA-polymer conjugates are often limited by low reaction yields due to the sterically hindered coupling of a presynthesized polymer to DNA. The grafting-from strategy, in contrast, allows one to directly graft polymers from an initiator that is covalently attached to DNA. Herein, we report blue-light-mediated reversible addn.-fragmentation chain-transfer (Photo-RAFT) polymn. from two different RAFT agent-terminated DNA sequences using Eosin Y as the photocatalyst in combination with ascorbic acid. Three monomer families (methacrylates, acrylates and acrylamides) were successfully polymd. from DNA employing Photo-RAFT polymn. We demonstrate that the length of the grown polymer chain can be varied by altering the monomer to DNA-initiator ratio, while the self-assembly features of the DNA strands were maintained. In summary, we describe a convenient, light-mediated approach toward DNA-polymer conjugates via the grafting-from approach.
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    Niu, J.; Lunn, D. J.; Pusuluri, A.; Yoo, J. I.; O’Malley, M. A.; Mitragotri, S.; Soh, H. T.; Hawker, C. J. Engineering Live Cell Surfaces with Functional Polymers via Cytocompatible Controlled Radical Polymerization. Nat. Chem. 2017, 9, 537– 545,  DOI: 10.1038/nchem.2713
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    Engineering live cell surfaces with functional polymers via cytocompatible controlled radical polymerization
    Niu, Jia; Lunn, David J.; Pusuluri, Anusha; Yoo, Justin I.; O'Malley, Michelle A.; Mitragotri, Samir; Soh, H. Tom; Hawker, Craig J.
    Nature Chemistry (2017), 9 (6), 537-545CODEN: NCAHBB; ISSN:1755-4330. (Nature Publishing Group)
    The capability to graft synthetic polymers onto the surfaces of live cells offers the potential to manipulate and control their phenotype and underlying cellular processes. Conventional grafting-to strategies for conjugating preformed polymers to cell surfaces are limited by low polymer grafting efficiency. Here we report an alternative grafting-from strategy for directly engineering the surfaces of live yeast and mammalian cells through cell surface-initiated controlled radical polymn. By developing cytocompatible PET-RAFT (photoinduced electron transfer-reversible addn.-fragmentation chain-transfer polymn.), synthetic polymers with narrow polydispersity (Mw/Mn < 1.3) could be obtained at room temp. in 5 min. This polymn. strategy enables chain growth to be initiated directly from chain-transfer agents anchored on the surface of live cells using either covalent attachment or non-covalent insertion, while maintaining high cell viability. Compared with conventional grafting-to approaches, these methods significantly improve the efficiency of grafting polymer chains and enable the active manipulation of cellular phenotypes.
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    Li, M.; Fromel, M.; Ranaweera, D.; Rocha, S.; Boyer, C.; Pester, C. W. SI-PET-RAFT: Surface-Initiated Photoinduced Electron Transfer-Reversible Addition–Fragmentation Chain Transfer Polymerization. ACS Macro Lett. 2019, 8, 374– 380,  DOI: 10.1021/acsmacrolett.9b00089
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    SI-PET-RAFT: Surface-Initiated Photoinduced Electron Transfer-Reversible Addition-Fragmentation Chain Transfer Polymerization
    Li, Mingxiao; Fromel, Michele; Ranaweera, Dhanesh; Rocha, Sergio; Boyer, Cyrille; Pester, Christian W.
    ACS Macro Letters (2019), 8 (4), 374-380CODEN: AMLCCD; ISSN:2161-1653. (American Chemical Society)
    In this communication, surface-initiated photoinduced electron transfer-reversible addn.-fragmentation chain transfer polymn. (SI-PET-RAFT) is introduced. SI-PET-RAFT affords functionalization of surfaces with spatiotemporal control and provides oxygen tolerance under ambient conditions. All hallmarks of controlled radical polymn. (CRP) are met, affording well-defined polymn. kinetics, and chain end retention to allow subsequent extension of active chain ends to form block copolymers. The modularity and versatility of SI-PET-RAFT is highlighted through significant flexibility with respect to the choice of monomer, light source and wavelength, and photoredox catalyst. The ability to obtain complex patterns in the presence of air is a significant contribution to help pave the way for CRP-based surface functionalization into com. application.
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    Kuzmyn, A. R.; Teunissen, L. W.; Fritz, P.; van Lagen, B.; Smulders, M. M. J.; Zuilhof, H. Diblock and Random Antifouling Bioactive Polymer Brushes on Gold Surfaces by Visible-Light-Induced Polymerization (SI-PET-RAFT) in Water. Adv. Mater. Interfaces 2022, 9, 2101784  DOI: 10.1002/admi.202101784
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    46
    Diblock and random antifouling bioactive polymer brushes on gold surfaces by visible-light-induced polymerization in water
    Kuzmyn, Andriy R.; Teunissen, Lucas W.; Fritz, Pina; van Lagen, Barend; Smulders, Maarten M. J.; Zuilhof, Han
    Advanced Materials Interfaces (2022), 9 (3), 2101784CODEN: AMIDD2; ISSN:2196-7350. (Wiley-VCH Verlag GmbH & Co. KGaA)
    Surface-initiated photoinduced electron-transfer-reversible addn.-fragmentation chain transfer (SI-PET-RAFT) is, for the first time, used for the creation of antifouling polymer brushes on gold surfaces based on three monomers: oligo(ethylene glycol) Me ether methacrylate (MeOEGMA), N-(2-hydroxypropyl) methacrylamide (HPMA), and carboxybetaine methacrylamide (CBMA). These coatings are subsequently characterized by XPS and ellipsometry. The living nature of this polymn. allows for the creation of random and diblock copolymer brushes, which are based on HPMA (superb antifouling) and CBMA (good antifouling and functionalizable via activated ester chem.). The polymer brushes demonstrate good antifouling properties against undiluted human serum, as monitored by quartz crystal microbalance with dissipation (QCM-D) and surface plasmon resonance (SPR) spectroscopy in real time. The amt. of immobilization of bioactive moieties, here an antibody immobilized using N-succinimidyl ester-1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (NHS-EDC) coupling, in the diblock and random copolymer brushes is monitored by SPR, and is analyzed with respect to the brush structure, and is shown to be superior in the diblock copolymer brush. This approach represents a scalable, robust, mild, oxygen-tolerant, and heavy-metal-free route toward the prodn. of antifouling and functional copolymer brushes (on gold surfaces) that open up applications in biosensing and tissue engineering.
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    Roeven, E.; Kuzmyn, A. R.; Scheres, L.; Baggerman, J.; Smulders, M. M. J.; Zuilhof, H. PLL–Poly(HPMA) Bottlebrush-Based Antifouling Coatings: Three Grafting Routes. Langmuir 2020, 36, 10187– 10199,  DOI: 10.1021/acs.langmuir.0c01675
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    PLL-Poly(HPMA) Bottlebrush-Based Antifouling Coatings: Three Grafting Routes
    Roeven, Esther; Kuzmyn, Andriy R.; Scheres, Luc; Baggerman, Jacob; Smulders, Maarten M. J.; Zuilhof, Han
    Langmuir (2020), 36 (34), 10187-10199CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    In this work, we compare three routes to prep. antifouling coatings that consist of poly(L-lysine)-poly(N-(2-hydroxypropyl)methacrylamide) bottlebrushes. The poly(L-lysine) (PLL) backbone is self-assembled onto the surface by charged-based interactions between the lysine groups and the neg. charged silicon oxide surface, whereas the poly(N-(2-hydroxypropyl)methacrylamide) [poly(HPMA)] side chains, grown by reversible addn.-fragmentation chain-transfer (RAFT) polymn., provide antifouling properties to the surface. First, the PLL-poly(HPMA) coatings are synthesized in a bottom-up fashion through a grafting-from approach. In this route, the PLL is self-assembled onto a surface, after which a polymn. agent is immobilized, and finally HPMA is polymd. from the surface. In the second explored route, the PLL is modified in soln. by a RAFT agent to create a macroinitiator. After self-assembly of this macroinitiator onto the surface, poly(HPMA) is polymd. from the surface by RAFT. In the third and last route, the whole PLL-poly(HPMA) bottlebrush is initially synthesized in soln. To this end, HPMA is polymd. from the macroinitiator in soln. and the PLL-poly(HPMA) bottlebrush is then self-assembled onto the surface in just one step (grafting-to approach). Addnl., in this third route, we also design and synthesize a bottlebrush polymer with a PLL backbone and poly(HPMA) side chains, with the latter contg. 5% carboxybetaine (CB) monomers that eventually allow for addnl. (bio)functionalization in soln. or after surface immobilization. These three routes are evaluated in terms of ease of synthesis, scalability, ease of characterization, and a preliminary investigation of their antifouling performance. All three coating procedures result in coatings that show antifouling properties in single-protein antifouling tests. This method thus presents a new, simple, versatile, and highly scalable approach for the manufg. of PLL-based bottlebrush coatings that can be synthesized partly or completely on the surface or in soln., depending on the desired prodn. process and/or application.
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    Xu, J.; Shanmugam, S.; Duong, H. T.; Boyer, C. Organo-photocatalysts for Photoinduced Electron Transfer-reversible Addition–fragmentation Chain Transfer (PET-RAFT) Polymerization. Polym. Chem. 2015, 6, 5615– 5624,  DOI: 10.1039/C4PY01317D
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    Organo-photocatalysts for photoinduced electron transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization
    Xu, Jiangtao; Shanmugam, Sivaprakash; Duong, Hien T.; Boyer, Cyrille
    Polymer Chemistry (2015), 6 (31), 5615-5624CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
    In this article, we investigated a series of organo-dyes, including methylene blue, fluorescein, Rhodamine 6G, Nile red and eosin Y, to perform a visible light-mediated controlled/"living" radical polymn. of methacrylates. We demonstrate that eosin Y and fluorescein were efficient catalysts to activate a photoinduced electron transfer-reversible addn.-fragmentation chain transfer (PET-RAFT) mechanism. The concn. of eosin Y was varied from 10 to 100 ppm with respect to monomers. This polymn. technique yielded well-defined (co)polymers with a good control of the mol. wts. ranging from 10 000 to 100 000 g mol-1 and low polydispersities (PDI < 1.30). A variety of functional monomers, including N,N-dimethylaminoethyl methacrylate, hydroxyl Et methacrylate, pentafluorophenyl methacrylate, glycidyl methacrylate, oligo(ethylene glycol) Me ether methacrylate (OEGMA), and methacrylic acid, were successfully polymd. Finally, the addn. of a tertiary amine, such as triethylamine, afforded the polymn. in the presence of air via a reductive quenching cycle. Different diblock polymethacrylate copolymers, i.e.PMMA-b-POEGMA and PMMA-b-PMMA, were prepd. to demonstrate the high end group fidelity.
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    Cheng, G.; Li, G.; Xue, H.; Chen, S.; Bryers, J. D.; Jiang, S. Zwitterionic Carboxybetaine Polymer Surfaces and their Resistance to Long-term Biofilm Formation. Biomaterials 2009, 30, 5234– 5240,  DOI: 10.1016/j.biomaterials.2009.05.058
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    Zwitterionic carboxybetaine polymer surfaces and their resistance to long-term biofilm formation
    Cheng, Gang; Li, Guozhu; Xue, Hong; Chen, Shengfu; Bryers, James D.; Jiang, Shaoyi
    Biomaterials (2009), 30 (28), 5234-5240CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
    The authors report a systematic study of zwitterionic poly(carboxybetaine methacrylate) (pCBMA) grafted from glass surfaces via atom transfer radical polymn. (ATRP) for their resistance to long-term bacterial biofilm formation. Results show that pCBMA-grafted surfaces are highly resistant to non-specific protein adsorption (fibrinogen and undiluted blood plasma) at 25, 30 and 37°. Long-term (over 24 h) colonization of two bacterial strains (Pseudomonas aeruginosa PAO1 and Pseudomonas putida strain 239) on pCBMA surface was studied using a parallel flow cell at 25, 30 and 37°. Uncoated glass cover slips were chosen as the pos. ref. Results show that pCBMA coatings reduced long-term biofilm formation of P. aeruginosa up to 240 h by 95% at 25° and for 64 h by 93% at 37°, and suppressed P. putida biofilm accumulation up to 192 h by 95% at 30°, with respect to the glass ref. The ability of pCBMA coatings to resist non-specific protein adsorption and significantly retard bacterial biofilm formation makes it a very promising material for biomedical and industrial applications.
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    van Andel, E.; Lange, S. C.; Pujari, S. P.; Tijhaar, E. J.; Smulders, M. M. J.; Savelkoul, H. F. J.; Zuilhof, H. Systematic Comparison of Zwitterionic and Non-Zwitterionic Antifouling Polymer Brushes on a Bead-Based Platform. Langmuir 2019, 35, 1181– 1191,  DOI: 10.1021/acs.langmuir.8b01832
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    Systematic Comparison of Zwitterionic and Non-Zwitterionic Antifouling Polymer Brushes on a Bead-Based Platform
    van Andel, Esther; Lange, Stefanie C.; Pujari, Sidharam P.; Tijhaar, Edwin J.; Smulders, Maarten M. J.; Savelkoul, Huub F. J.; Zuilhof, Han
    Langmuir (2019), 35 (5), 1181-1191CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    Nonspecific adsorption of biomols. to solid surfaces, a process called biofouling, is a major concern in many biomedical applications. Great effort has been made in the development of antifouling polymer coatings that are capable of repelling the nonspecific adsorption of proteins, cells, and micro-organisms. In this respect, we herein contribute to understanding the factors that det. which polymer brush results in the best antifouling coating. To this end, we compared five different monomers: two sulfobetaines, a carboxybetaine, a phosphocholine, and a hydroxyl acrylamide. The antifouling coatings were analyzed using our previously described bead-based method with flow cytometry as the read-out system. This method allows for the quick and automated anal. of thousands of beads per s, enabling fast anal. and good statistics. We report the first direct comparison made between a sulfobetaine with opposite charges sepd. by two and three methylene groups and a carboxybetaine bearing two sepg. methylene groups. It was concluded that both the distance between opposite charges and the nature of the anionic groups have a distinct effect on the antifouling performance. Phosphocholines and simple hydroxyl acrylamides are not often compared with the betaines. However, here we found that they perform equally well or even better, yielding the following overall antifouling ranking: HPMAA ≥ PCMA-2 ≈ CBMAA-2 > SBMAA-2 > SBMAA-3 » nonmodified beads (HPMAA being the best).
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    Sah, P.; Alfaro-Murillo, J. A.; Fitzpatrick, M. C.; Neuzil, K. M.; Meyers, L. A.; Singer, B. H.; Galvani, A. P. Future Epidemiological and Economic Impacts of Universal Influenza Vaccines. Proc. Natl. Acad. Sci. U.S.A. 2019, 116, 20786– 20792,  DOI: 10.1073/pnas.1909613116
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    Future epidemiological and economic impacts of universal influenza vaccines
    Sah, Pratha; Alfaro-Murillo, Jorge A.; Fitzpatrick, Meagan C.; Neuzil, Kathleen M.; Meyers, Lauren A.; Singer, Burton H.; Galvani, Alison P.
    Proceedings of the National Academy of Sciences of the United States of America (2019), 116 (41), 20786-20792CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
    The efficacy of influenza vaccines, currently at 44%, is limited by the rapid antigenic evolution of the virus and a manufg. process that can lead to vaccine mismatch. The National Institute of Allergy and Infectious Diseases (NIAID) recently identified the development of a universal influenza vaccine with an efficacy of at least 75% as a high scientific priority. The US Congress approved $130 million funding for the 2019 fiscal year to support the development of a universal vaccine, and another $1 billion over 5 y has been proposed in the Flu Vaccine Act. Using a model of influenza transmission, we evaluated the population-level impacts of universal influenza vaccines distributed according to empirical age-specific coverage at multiple scales in the United States. We est. that replacing just 10% of typical seasonal vaccines with 75% efficacious universal vaccines would avert ∼5.3 million cases, 81,000 hospitalizations, and 6,300 influenza-related deaths per yr. This would prevent over $1.1 billion in direct health care costs compared to a typical season, based on av. data from the 2010-11 to 2018-19 seasons. A complete replacement of seasonal vaccines with universal vaccines is projected to prevent 17 million cases, 251,000 hospitalizations, 19,500 deaths, and $3.5 billion in direct health care costs. States with high per-hospitalization medical expenses along with a large proportion of elderly residents are expected to receive the max. economic benefit. Replacing even a fraction of seasonal vaccines with universal vaccines justifies the substantial cost of vaccine development.
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    Vaisocherová, H.; Ševců, V.; Adam, P.; Špačková, B.; Hegnerová, K.; de los Santos Pereira, A.; Rodriguez-Emmenegger, C.; Riedel, T.; Houska, M.; Brynda, E.; Homola, J. Functionalized Ultra-low Fouling Carboxy- and Hydroxy-functional Surface Platforms: Functionalization Capacity, Biorecognition Capability and Resistance to Fouling from Undiluted Biological Media. Biosens. Bioelectron. 2014, 51, 150– 157,  DOI: 10.1016/j.bios.2013.07.015
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    Functionalized ultra-low fouling carboxy- and hydroxy-functional surface platforms: functionalization capacity, biorecognition capability and resistance to fouling from undiluted biological media
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    Biosensors & Bioelectronics (2014), 51 (), 150-157CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
    The non-specific binding of non-target species to functionalized surfaces of biosensors continues to be challenge for biosensing in real-world media. Three different low-fouling and functionalizable surface platforms were employed to study the effect of functionalization on fouling resistance from several types of undiluted media including blood plasma and food media. The surface platforms investigated in this work included two polymer brushes: hydroxy-functional poly(2-hydroxyethyl methacrylate) (pHEMA) and carboxy-functional poly(carboxybetaine acrylamide) (pCBAA), and a std. OEG-based carboxy-functional alkanethiolate self-assembled monolayer (AT-SAM). The wet and dry polymer brushes were analyzed by AFM, ellipsometry, FT-IRRAS, and surface plasmon resonance (SPR). The surfaces were functionalized by the covalent attachment of antibodies, streptavidin, and oligonucleotides and the binding and biorecognition characteristics of the coatings were compared. We found that functionalization did not substantially affect the ultra-low fouling properties of pCBAA (plasma fouling of ∼20 ng/cm2), a finding in contrast with pHEMA that completely lost its resistance to fouling after the activation of hydroxyl groups. Blocking a functionalized AT-SAM covalently with BSA decreased fouling down to the level comparable to unblocked pCBAA. However, the biorecognition capability of blocked functionalized AT-SAM was poor in comparison with functionalized pCBAA. Limits of detection of Escherichia coli O157:H7 in undiluted milk were detd. to be 6×104, 8×105, and 6×105 cells/mL for pCBAA, pHEMA, and AT-SAM-blocked, resp. Effect of analyte size on biorecognition activity of functionalized coatings was investigated and it was shown that the best performance in terms of overall fouling resistance and biorecognition capability is provided by pCBAA.
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    Vaisocherová-Lísalová, H.; Surman, F.; Víšová, I.; Vala, M.; Špringer, T.; Ermini, M. L.; Šípová, H.; Šedivák, P.; Houska, M.; Riedel, T.; Pop-Georgievski, O.; Brynda, E.; Homola, J. Copolymer Brush-Based Ultralow-Fouling Biorecognition Surface Platform for Food Safety. Anal. Chem. 2016, 88, 10533– 10539,  DOI: 10.1021/acs.analchem.6b02617
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    Copolymer Brush-Based Ultralow-Fouling Biorecognition Surface Platform for Food Safety
    Vaisocherova-Lisalova, Hana; Surman, Frantisek; Visova, Ivana; Vala, Milan; Springer, Tomas; Ermini, Maria Laura; Sipova, Hana; Sedivak, Petr; Houska, Milan; Riedel, Tomas; Pop-Georgievski, Ognen; Brynda, Eduard; Homola, Jiri
    Analytical Chemistry (Washington, DC, United States) (2016), 88 (21), 10533-10539CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
    Functional polymer coatings that combine the ability to resist non-specific fouling from complex media with high biorecognition element (BRE) immobilization capacity represent an emerging class of new functional materials for a no. of bioanal. and biosensor technologies for medical diagnostics, security, and food safety. Here we report on a random copolymer brush surface - poly(CBMAA-ran-HPMAA) - providing high BRE immobilization capacity while simultaneously exhibiting ultralow-fouling behavior in complex food media. We demonstrate that both the functionalization and fouling resistance capabilities of such copolymer brushes can be tuned by changing the surface contents of the two monomer units: non-ionic N-(2-hydroxypropyl) methacrylamide (HPMAA) and carboxy-functional zwitterionic carboxybetaine methacrylamide (CBMAA). It is demonstrated that the resistance to fouling decreases with the surface content of CBMAA; poly(CBMAA-ran-HPMAA) brushes with CBMAA molar content up to 15 mol% maintain excellent resistance to fouling from a variety of homogenized foods (hamburger, cucumber, milk, and lettuce) even after covalent attachment of BREs to carboxy-groups of CBMAA. The poly(CBMAA 15 mol%-ran-HPMAA) brushes functionalized with antibodies are demonstrated to exhibit fouling resistance from food samples by up to three orders of magnitude better when compared with the widely used low-fouling carboxy-functional oligo(ethylene glycol) (OEG)-based alkanethiolate self-assembled monolayers (AT SAMs) and furthermore, by up to two orders of magnitude better when compared with the most successful ultralow-fouling biorecognition coatings - poly(carboxybetaine acrylamide), poly(CBAA). Using model SPR detections of foodborne bacterial pathogens in homogenized foods it is also demonstrated that the antibody-functionalized poly(CBMAA 15 mol%-ran-HPMAA) brush exhibits superior biorecognition properties over the poly(CBAA).
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    A review. XPS is widely used to identify and quantify the elements present at the surface of polymeric materials. The energy distribution of photoelectrons emitted from these elements contains information about their chem. state, potentially allowing the analyst to identify and quantify specific functional groups. These functional groups may originate from the synthesis and processing of the polymers, from postsynthetic modifications such as surface grafting, or indeed may be unrelated to the polymer (additives and contaminants). Extg. reliable and meaningful information from XPS data is not trivial and relies on careful and appropriate experimentation, including exptl. design, sample prepn., data collection, data processing, and data interpretation. Here, the authors outline some of these challenges when performing XPS anal. of polymers and provide practical examples to follow. This guide will cover all relevant aspects over the course of a typical expt., including tips and considerations when designing the expt., sample prepn., charge neutralization, x-ray induced sample damage, depth profiling, data anal. and interpretation, and, finally, reporting of results. Many of these topics are more widely applicable to insulating org. materials, and the recommendations of this guide will help to ensure that data is collected and interpreted using current best practices. (c) 2020 American Institute of Physics.
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    Spread of highly pathogenic avian influenza (HPAI) H5N5 viruses in europe in 2016-2017 appears related to the timing of reassortment events
    Bergervoet, Saskia A.; Ho, Cynthia K. Y.; Heutink, Rene; Bossers, Alex; Beerens, Nancy
    Viruses (2019), 11 (6), 501CODEN: VIRUBR; ISSN:1999-4915. (MDPI AG)
    During the epizootic of highly pathogenic avian influenza (HPAI) H5N8 virus in Europe in 2016-2017, HPAI viruses of subtype H5N5 were also isolated. However, the detection of H5N5 viruses was limited compared to H5N8. In this study, we show that the genetic constellation of a newly isolated H5N5 virus is different from two genotypes previously identified in the Netherlands. The introduction and spread of the three H5N5 genotypes in Europe was studied using spatiotemporal and genetic anal. This demonstrated that the genotypes were isolated in distinguishable phases of the epizootic, and suggested multiple introductions of H5N5 viruses into Europe followed by local spread. We estd. the timing of the reassortment events, which suggested that the genotypes emerged after the start of autumn migration. This may have prevented large-scale spread of the H5N5 viruses on wild bird breeding sites before introduction into Europe. Expts. in primary chicken and duck cells revealed only minor differences in cytopathogenicity and replication kinetics between H5N5 genotypes and H5N8. These results suggest that the limited spread of HPAI H5N5 viruses is related to the timing of the reassortment events rather than changes in virus pathogenicity or replication kinetics.
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    Predictive Value of Precision-Cut Lung Slices for the Susceptibility of Three Animal Species for SARS-CoV-2 and Validation in a Refined Hamster Model
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    Pathogens (2021), 10 (7), 824CODEN: PATHCD; ISSN:2076-0817. (MDPI AG)
    In assessing species susceptibility for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and in the search for an appropriate animal model, multiple research groups around the world inoculated a broad range of animal species using various SARS-CoV-2 strains, doses and administration routes. Although in silico analyses based on receptor binding and diverse in vitro cell cultures were valuable, exact prediction of species susceptibility based on these tools proved challenging. Here, we assessed whether precision-cut lung slices (PCLS) could facilitate the selection of animal models, thereby reducing animal experimentation. Pig, hamster and cat PCLS were incubated with SARS-CoV-2 and virus replication was followed over time. Virus replicated efficiently in PCLS from hamsters and cats, while no evidence of replication was obtained for pig PCLS. These data corroborate the findings of many research groups that have investigated the susceptibility of hamsters, pigs and cats towards infection with SARS-CoV-2. Our findings suggest that PCLS can be used as convenient tool for the screening of different animal species for sensitivity to newly emerged viruses. To validate our results obtained in PCLS, we employed the hamster model. Hamsters were inoculated with SARS-CoV-2 via the intranasal route. Susceptibility to infection was evaluated by body wt. loss, viral loads in oropharyngeal swabs and respiratory tissues and lung pathol. The broadly used hamster model was further refined by including activity tracking of the hamsters by an activity wheel as a very robust and sensitive parameter for clin. health. In addn., to facilitate the quantification of pathol. in the lungs, we devised a semi-quant. scoring system for evaluating the degree of histol. changes in the lungs. The inclusion of these addnl. parameters refined and enriched the hamster model, allowing for the generation of more data from a single expt.
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    Multi-functional initiator and poly(carboxybetaine methacrylamides) for building biocompatible surfaces using "nitroxide mediated free radical polymerization" strategies
    Abraham, Sinoj; Unsworth, Larry D.
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    Biomaterials generally suffer from rapid nonspecific protein adsorption, which initiates many deleterious host responses, and complex chemistries that are employed to facilitate cellular interactions. A chem. approach that, based upon current literature, combines a nonfouling architecture with a biomemtic cell-adhesive end-group, is presented. Namely, surface-initiated polymn. of zwitterionic [poly (carboxybetaine methacrylamide)] brushes, with controlled charge densities and phosphonate head groups. Nitroxide mediated free radical polymn. (NMFRP) was employed for various reasons: reduces presence of potentially cytotoxic organometallic catalysts common in atom transfer radical polymn. (ATRP); and it allows a phosphonate end-group instead of the common brominated end-group. Thermally oxidized silicon wafers were covalently functionalized with diethyl-(1-(N-(1-(3-(trimethoxysilyl)propylcarbamoyl)ethoxy)-N-tert-butylamino)ethyl)phosphonate. NMFRP was used to graft zwitterionic carboxybetaine methacrylamide monomers of varying inter-charge sepn. The resulting thin films were characterized using Attenuated Total Reflectance-Fourier Transform IR (ATR-FTIR) and X-ray photoelectron (XPS) spectroscopy, ellipsometry, water contact angle anal., and thermo gravimetric anal. (TGA). The effect of spacer group on the surface charge d. was detd. using zeta potential techniques. It is thought that this stratagem will facilitate the ability to tailor systematically both the interior and terminal polymer properties, providing a platform for further understanding how these conditions affect protein adsorption as well as cell-surface interactions. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011.
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    Giesbers, M.; Marcelis, A. T. M.; Zuilhof, H. Simulation of XPS C1s Spectra of Organic Monolayers by Quantum Chemical Methods. Langmuir 2013, 29, 4782– 4788,  DOI: 10.1021/la400445s
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    Simulation of XPS C1s Spectra of Organic Monolayers by Quantum Chemical Methods
    Giesbers, Marcel; Marcelis, Antonius T. M.; Zuilhof, Han
    Langmuir (2013), 29 (15), 4782-4788CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    Several simple methods are presented and evaluated to simulate the X-ray photoelectron spectra (XPS) of org. monolayers and polymeric layers by d. functional theory (DFT) and second-order Moller-Plesset theory (MP2) in combination with a series of basis sets. The simulated carbon (C1s) XPS spectra as obtained via B3LYP/6-311G(d,p) or M11/6-311G(d,p) calcns. are in good agreement (av. mean error <0.3 eV) with the exptl. spectra, and good ests. of C1s spectra can be obtained via EC1s(exp) = 0.9698EC1s(theory) + 20.34 (in eV) (B3LYP/6-311G(d,p)). As a result, the simulated C1s XPS spectra can elucidate the binding energies of the different carbon species within an org. layer and, in this way, greatly aid the assignment of complicated C1s XPS spectra. A wide range of examples, including haloalkanes, esters, (thio-)ethers, leaving groups, clickable functionalities, and bioactive moieties, are given.
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    Zhao, J.; Gao, F.; Pujari, S. P.; Zuilhof, H.; Teplyakov, A. V. Universal Calibration of Computationally Predicted N 1s Binding Energies for Interpretation of XPS Experimental Measurements. Langmuir 2017, 33, 10792– 10799,  DOI: 10.1021/acs.langmuir.7b02301
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    Universal Calibration of Computationally Predicted N 1s Binding Energies for Interpretation of XPS Experimental Measurements
    Zhao, Jing; Gao, Fei; Pujari, Sidharam P.; Zuilhof, Han; Teplyakov, Andrew V.
    Langmuir (2017), 33 (41), 10792-10799CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    Computationally predicted N 1s core level energies are commonly used to interpret the exptl. measurements obtained with XPS. This work compares the application of Koopmans' theorem to core electrons using the B3LYP functional with 2 commonly used basis sets, analyzes the factors relevant to the comparison of the computational with exptl. data, and presents several correlations that allow an accurate prediction of the N 1s binding energy. The 1st correlation is obtained with known N-contg. functional groups on well-characterized org. monolayers. This approach can then be reliably extended to a no. of N-contg. chem. systems on Si surfaces in which the nature of the chem. environment of N atoms had only been proposed based on a no. of anal. techniques. In most of those cases, the XPS anal. is consistent with the proposed structures, but is not always sufficient for conclusive assignments. Third, it was attempted to also include N-contg. systems on metals. Despite the admittedly oversimplified approach taken in this case (the metal surface is approximated by a single atom), the obsd. correlations are still exptl. useful, although in this case significant outliers are found. Finally, previously published correlations between exptl. and theor. C 1s data were reexamd., yielding a set of correlations that allow experimentalists to predict C 1s and N 1s XPS spectra with high accuracy.
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    Zhang, L.; Cao, Z.; Bai, T.; Carr, L.; Ella-Menye, J.-R.; Irvin, C.; Ratner, B. D.; Jiang, S. Zwitterionic hydrogels implanted in mice resist the foreign-body reaction. Nat. Biotechnol. 2013, 31, 553– 556,  DOI: 10.1038/nbt.2580
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    Zwitterionic hydrogels implanted in mice resist the foreign-body reaction
    Zhang, Lei; Cao, Zhiqiang; Bai, Tao; Carr, Louisa; Ella-Menye, Jean-Rene; Irvin, Colleen; Ratner, Buddy D.; Jiang, Shaoyi
    Nature Biotechnology (2013), 31 (6), 553-556CODEN: NABIF9; ISSN:1087-0156. (Nature Publishing Group)
    The performance of implantable biomedical devices is impeded by the foreign-body reaction, which results in formation of a dense collagenous capsule that blocks mass transport and/or elec. communication between the implant and the body. No known materials or coatings can completely prevent capsule formation. Here we demonstrate that ultra-low-fouling zwitterionic hydrogels can resist the formation of a capsule for at least 3 mo after s.c. implantation in mice. Zwitterionic hydrogels also promote angiogenesis in surrounding tissue, perhaps owing to the presence of macrophages exhibiting phenotypes assocd. with anti-inflammatory, pro-healing functions. Thus, zwitterionic hydrogels may be useful in a broad range of applications, including generation of biocompatible implantable medical devices and tissue scaffolds.
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    Tiwari, A.; Devi, P. P.; Yogesh, C.; Minakshi, P.; Sagar, M. G. Survival of Two Avian Respiratory Viruses on Porous and Nonporous Surfaces. Avian Dis. 2006, 50, 284– 287,  DOI: 10.1637/7453-101205R.1
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    Survival of two avian respiratory viruses on porous and nonporous surfaces
    Tiwari A; Patnayak Devi P; Chander Yogesh; Parsad Minakshi; Goyal Sagar M
    Avian diseases (2006), 50 (2), 284-7 ISSN:0005-2086.
    The transmission of pathogens from infected to susceptible hosts may occur through contaminated fomites and inanimate objects. This type of transmission depends on the ability of the pathogens to survive in the environment. In this report, we describe the survivability of two avian respiratory viruses, e.g., avian metapneumovirus and avian influenza virus on 12 different porous and nonporous surfaces. The viruses survived on some of the surfaces for up to 6 days postcontamination but not after 9 days. Both viruses survived longer on nonporous surfaces than on porous ones. One of the reasons for poor survival on porous surfaces could be inefficient elution of virus from these surfaces. These results should be helpful in determining how long the premises should be left vacant after an outbreak of these viruses has occurred in poultry houses.
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  • Abstract

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    Scheme 1

    Scheme 1. (Top) General Scheme of Polymer Brush-Based Antiviral Surfaces; (Bottom) Structure of Monomers A10 and CBMA Used to Construct Quaternary Ammonium and Zwitterionic Polymer Brushes
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    Scheme 2

    Scheme 2. General Scheme for the Synthesis of Antiviral Polymer Brushes
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    Figure 1

    Figure 1. Physicochemical properties of polymer brush-coated surfaces. XPS wide spectra of poly(CBMA)- and poly(A10)-coated surfaces (a). XPS N 1s narrow scan (b) and XPS C 1s narrow scan (c) of poly(CBMA)- and poly(A10)-coated surfaces. Typical AFM topography images of poly(A10)- (d) and poly(CBMA)-coated surfaces (e).

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    Figure 2

    Figure 2. Antiviral activity of copper-coated surfaces, and poly(CBMA) and poly(A10) brush surfaces against SARS-CoV-2 and avian influenza. Plasma-cleaned silicon surfaces were used as a negative control.

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    Scheme 3

    Scheme 3. General Scheme of the Suggested Antiviral Mechanism of Lipid Bilayer-Disrupting Polymer Brushes (1–3); Monomer Chemical Structure and Monomer Schematic Representation Are Shown Below
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      Langmuir (2020), 36 (16), 4439-4446CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      This work presents a new method for the synthesis of antifouling polymer brushes using surface-initiated photoinduced electron transfer-reversible addn.-fragmentation chain-transfer polymn. with eosin Y and triethanolamine as catalysts. This method proceeds in an aq. environment under atm. conditions without any prior degassing and without the use of heavy metal catalysts. The versatility of the method is shown by using three chem. different monomers: oligo(ethylene glycol) methacrylate, N-(2-hydroxypropyl)methacrylamide, and carboxybetaine methacrylamide. In addn., the light-triggered nature of the polymn. allows the creation of complex three-dimensional structures. The compn. and topol. structuring of the brushes are confirmed by XPS and at. force microscopy. The kinetics of the polymns. are followed by measuring the layer thickness with ellipsometry. The polymer brushes demonstrate excellent antifouling properties when exposed to single-protein solns. and complex biol. matrixes such as dild. bovine serum. This method thus presents a new simple approach for the manufg. of antifouling coatings for biomedical and biotechnol. applications.
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    22. 22
      Kuzmyn, A. R.; Nguyen, A. T.; Zuilhof, H.; Baggerman, J. Bioactive Antifouling Surfaces by Visible-Light-Triggered Polymerization. Adv. Mater. Interfaces 2019, 6, 1900351  DOI: 10.1002/admi.201900351
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      Pop-Georgievski, O.; Rodriguez-Emmenegger, C.; Pereira, A. d. l. S.; Proks, V.; Brynda, E.; Rypáček, F. Biomimetic Non-fouling Surfaces: Extending the Concepts. J. Mater. Chem. B 2013, 1, 2859– 2867,  DOI: 10.1039/c3tb20346h
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      23
      Biomimetic non-fouling surfaces: extending the concepts
      Pop-Georgievski, Ognen; Rodriguez-Emmenegger, Cesar; de los Santos Pereira, Andres; Proks, Vladimir; Brynda, Eduard; Rypacek, Frantisek
      Journal of Materials Chemistry B: Materials for Biology and Medicine (2013), 1 (22), 2859-2867CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)
      In this study, we propose a substrate-independent biomimetic modification route for the creation of antifouling polymer brushes. This modification route consists of the formation/deposition of a biomimetic polydopamine anchor layer followed by a well-controlled surface-initiated atom transfer radical polymn. of antifouling polymer brushes initiated by 2-bromo-2-methylpropanoyl groups covalently attached to the hydroxyl and amine groups present in the anchor layer. In this way, we synthesized polymer brushes of methoxy- and hydroxy-capped oligoethylene glycol methacrylate, 2-hydroxyethyl methacrylate and carboxybetaine acrylamide. Spectroscopic ellipsometry (SE) indicated well-controlled polymn. kinetics of the brushes, thus the thickness of the ultra-thin films could be precisely tuned at a nanometer scale. The covalent structure and organization of the brushes grown from the polydopamine anchor layer were accessed by IR reflection-adsorption spectroscopy (IRRAS) while the change in hydrophilicity caused by the presence of the brush was detd. by dynamic water contact angle measurements. Surface plasmon resonance as well as ex situ IRRAS and SE measurements were applied to investigate the adsorption of model protein solns. and undiluted human blood plasma to the brushes. The biomimetic brushes completely suppressed the fouling from single protein solns. and reduced the fouling from plasma to less than 3% from the fouling measured on bare gold surfaces. The proposed modification procedure is non-destructive and does not require any chem. pre-activation or the presence of reactive groups on the substrate surface. Contrary to other antifouling modifications the coating can be performed on various classes of substrates and preserves its properties even in undiluted blood plasma. This work offers a promising technol. for the facile fabrication of different surface-based biotechnol. and biomedical devices able to perform tailor-made functions while resisting the fouling from the complex biol. media where they operate.
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    24. 24
      Zamfir, M.; Rodriguez-Emmenegger, C.; Bauer, S.; Barner, L.; Rosenhahn, A.; Barner-Kowollik, C. Controlled Growth of Protein Resistant PHEMA Brushes via S-RAFT Polymerization. J. Mater. Chem. B 2013, 1, 6027– 6034,  DOI: 10.1039/c3tb20880j
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      Controlled growth of protein resistant PHEMA brushes via S-RAFT polymerization
      Zamfir, Mirela; Rodriguez-Emmenegger, Cesar; Bauer, Stella; Barner, Leonie; Rosenhahn, Axel; Barner-Kowollik, Christopher
      Journal of Materials Chemistry B: Materials for Biology and Medicine (2013), 1 (44), 6027-6034CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)
      The reversible addn.-fragmentation chain transfer polymn. of 2-hydroxyethyl methacrylate (HEMA) from surfaces (S-RAFT) using an R-group-attached chain transfer agent (CTA) is presented. The approach was exploited for the efficient prepn. of well-defined PHEMA brushes of up to 50 nm thickness in a controlled fashion without using any cytotoxic catalyst. The chem. compn., morphol. and wettability of the samples were assessed by XPS, at. force microscopy and water contact angle measurements, while the growth kinetics were studied by monitoring the dry thickness via spectroscopic ellipsometry. The mechanism and kinetics of the RAFT polymn. on the surface - in the presence of a sacrificial CTA and of solvent mixts. with different polarities - were investigated. A marked effect of the concn. of the sacrificial CTA on the kinetics was obsd. Importantly - and for the first time - the living PHEMA brushes were exploited as macroRAFT agents for chain extension, and thicknesses up to 70 nm were achieved. The prepd. PHEMA brushes were challenged with protein solns. demonstrating their resistance to fouling.
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    25. 25
      Lange, S. C.; van Andel, E.; Smulders, M. M. J.; Zuilhof, H. Efficient and Tunable Three-Dimensional Functionalization of Fully Zwitterionic Antifouling Surface Coatings. Langmuir 2016, 32, 10199– 10205,  DOI: 10.1021/acs.langmuir.6b02622
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      25
      Efficient and Tunable Three-Dimensional Functionalization of Fully Zwitterionic Antifouling Surface Coatings
      Lange, Stefanie C.; van Andel, Esther; Smulders, Maarten M. J.; Zuilhof, Han
      Langmuir (2016), 32 (40), 10199-10205CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      In order to enhance the sensitivity and selectivity of surface-based (bio)sensors, it is of crucial importance to diminish background signals that arise from the nonspecific binding of biomols., so-called biofouling. Zwitterionic polymer brushes have been shown to be excellent antifouling materials. However, for sensing purposes, antifouling does no suffice, but need to be combined with the possibility to efficiently modify the brush with recognition units. So far this has only been achieved at the expense of either antifouling properties or binding capacity. Herein the authors present a conceptually new approach by integrating both characteristics into a single, tailor-made monomer: a novel sulfobetaine-based zwitterionic monomer equipped with a clickable azide moiety. Copolymn. of this monomer with a well-established std. sulfobetaine monomer, results in highly antifouling surface coatings with a high, yet tunable amt. of clickable groups present throughout the entire brush. Subsequent functionalization of the azido-brushes, via e.g. widely used strain-promoted alkyne azide click reactions yields fully zwitterionic 3D-functionalized coatings with a recognition unit of choice that can be tailored for any specific application. Here the authors show a proof-of-principle with biotin-functionalized brushes on Si3N4 that combine excellent antifouling properties with specific avidin binding from a protein mixt. The signal-to-noise ratio is significantly improved over traditional chain end modification of sulfobetaine polymer brushes, even if the azide content is lowered to 1%, thus offering a viable approach for the development of significantly improved performance of biosensors on any surface.
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    26. 26
      Kuzmyn, A. R.; de los Santos Pereira, A.; Pop-Georgievski, O.; Bruns, M.; Brynda, E.; Rodriguez-Emmenegger, C. Exploiting End Group Functionalization for the Design of Antifouling Bioactive Brushes. Polym. Chem. 2014, 5, 4124– 4131,  DOI: 10.1039/C4PY00281D
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      Exploiting end group functionalization for the design of antifouling bioactive brushes
      Kuzmyn, A. R.; de los Santos Pereira, A.; Pop-Georgievski, O.; Bruns, M.; Brynda, E.; Rodriguez-Emmenegger, C.
      Polymer Chemistry (2014), 5 (13), 4124-4131CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
      Biol. active surfaces are essential in many applications in the fields of biosensing, bioimplants, and tissue engineering. However, the introduction of bioactive motifs without impairment of their ability to resist non-specific interactions with biol. media remains a challenge. Herein, we present a straightforward, facile strategy for the creation of bioactive surfaces based on the end-group biofunctionalization of state-of-the-art polymer brushes via an ultra-fast Diels-Alder "click" reaction. Surface-initiated atom transfer radical polymn. is employed to grow antifouling polymers preserving the end groups. These groups are then further converted to a reactive cyclopentadienyl moiety and exploited for the immobilization of biomols. on the topmost layer of the brush. The minimal chem. modification of the antifouling polymer brush accounts for the full preservation of the fouling resistance of the surface even after biofunctionalization, which is crit. for the aforementioned applications.
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      Rodriguez Emmenegger, C.; Brynda, E.; Riedel, T.; Sedlakova, Z.; Houska, M.; Alles, A. B. Interaction of Blood Plasma with Antifouling Surfaces. Langmuir 2009, 25, 6328– 6333,  DOI: 10.1021/la900083s
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      Interaction of blood plasma with antifouling surfaces
      Rodriguez Emmenegger C; Brynda E; Riedel T; Sedlakova Z; Houska M; Alles A Bologna
      Langmuir : the ACS journal of surfaces and colloids (2009), 25 (11), 6328-33 ISSN:0743-7463.
      Nonspecific adsorption of proteins is a crucial problem in the detection of analytes in complex biological media by affinity sensors operating with label-free detection. We modified the gold surface of surface plasmon resonance (SPR) sensors with three types of promising antifouling coatings: self-assembled monolayers (SAM)s of alkanethiolates terminated with diethylene glycol and carboxylic groups, poly(ethylene glycol) (PEG) grafted onto the SAMs, and zwitterionic polymer brushes of poly(carboxybetaine methacrylate), poly(sulfobetaine methacrylate), and poly(phosphorylcholine methacrylate). Using SPR, we compared the efficacy of the coatings to reduce nonspecific adsorption from human blood plasma and from single-protein solutions of human serum albumin, immunoglobulin G, fibrinogen, and lysozyme. There was no direct relationship between values of water contact angles and plasma deposition on the coated surfaces. A rather high plasma deposition on SAMs was decreased by grafting PEG chains. Fouling on PEG was observed only from plasma fractions containing proteins with molecular mass higher than 350 000 Da. The adsorption kinetics from plasma collected from different healthy donors differed. Poly(carboxybetaine methacrylate) completely prevented the deposition from plasma, but the other more hydrophilic zwitterionic polymers prevented single-protein adsorption but did not prevent plasma deposition. The results suggest that neither wettability nor adsorption of the main plasma proteins was the main indicator of deposition from blood plasma.
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      Williams, D. F. On the Nature of Biomaterials. Biomaterials 2009, 30, 5897– 5909,  DOI: 10.1016/j.biomaterials.2009.07.027
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      On the nature of biomaterials
      Williams, David F.
      Biomaterials (2009), 30 (30), 5897-5909CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
      A review. The situations in which biomaterials are currently used are vastly different to those of just a decade ago. Although implantable medical devices are still immensely important, medical technologies now encompass a range of drug and gene delivery systems, tissue engineering and cell therapies, organ printing and cell patterning, nanotechnol. based imaging and diagnostic systems and microelectronic devices. These technologies still encompass metals, ceramics and synthetic polymers, but also biopolymers, self assembled systems, nanoparticles, carbon nanotubes and quantum dots. These changes imply that our original concepts of biomaterials and our expectations of their performance also have to change. This Leading Opinion Paper addresses these issues. It concludes that many substances which hitherto we may not have thought of as biomaterials should now be considered as such so that, alongside the traditional structural biomaterials, we have substances that have been engineered to perform functions within health care where their performance is directly controlled by interactions with tissues and tissue components. These include engineered tissues, cells, organs and even viruses. This essay develops the arguments for a radically different definition of a biomaterial.
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    29. 29
      Rodriguez-Emmenegger, C.; Avramenko, O. A.; Brynda, E.; Skvor, J.; Alles, A. B. Poly(HEMA) Brushes Emerging as a New Platform for Direct Detection of Food Pathogen in Milk Samples. Biosens. Bioelectron. 2011, 26, 4545– 4551,  DOI: 10.1016/j.bios.2011.05.021
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      Poly(HEMA) brushes emerging as a new platform for direct detection of food pathogen in milk samples
      Rodriguez-Emmenegger, Cesar; Avramenko, Oxana A.; Brynda, Eduard; Skvor, J.; Alles, Aldo Bologna
      Biosensors & Bioelectronics (2011), 26 (11), 4545-4551CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
      Surface plasmon resonance (SPR) biosensors capable of in real time detection of Cronobacter at concns. down to 106 cells mL-1 in samples of consumer fresh-whole fat milk, powder whole-fat milk prepn., and powder infant formulation were developed for the first time. Antibodies against Cronobacter were covalently attached onto polymer brushes of poly(2-hydroxyethyl methacrylate) (poly(HEMA)) grafted from the SPR chip surface. The lowest detection limit, 104 cells mL-1, was achieved in phosphate buffered saline (pH 7.4) with sensors prepd. by covalent immobilization of the same antibodies onto a self assembled monolayer (SAM) of hexa(ethylene glycol) undecanethiol (EG6). However, when the EG6 based sensors were challenged with milk samples the non-specific response due to the deposition of non-targeted compds. from the milk samples was much higher than the specific response to Cronobacter hampering the detection in milk. Similar interfering fouling was obsd. on antifouling polymer brushes of hydroxy-capped oligoethylene glycol methacrylate and even a 10 times higher fouling was obsd. on the widely used SAM of mixed hydroxy- and carboxy-terminated alkanethiols. Only poly(HEMA) brushes totally suppressed the fouling from milk samples. The robust well-controlled surface initiated atom transfer radical polymn. of HEMA allowed the prepn. of highly dense brushes with a minimal thickness so that the capture of antigens by the antibodies immobilized on the brush layer could take place close to the gold SPR surface to provide a stronger optical response while the fouling was still suppressed. A min. thickness of 19 nm of poly(HEMA) brush layer was necessary to suppress completely non-specific sensor response to fouling from milk.
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      Liu, H.; Wang, S. Poly(N-isopropylacrylamide)-based thermo-responsive surfaces with controllable cell adhesion. Sci. China: Chem. 2014, 57, 552– 557,  DOI: 10.1007/s11426-013-5051-1
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      Poly(N-isopropylacrylamide)-based thermo-responsive surfaces with controllable cell adhesion
      Liu, Hong Liang; Wang, Shu Tao
      Science China: Chemistry (2014), 57 (4), 552-557CODEN: SCCCCS; ISSN:1869-1870. (Science China Press)
      A review. Poly(N-isopropylacrylamide) (PNIPAAm)-based thermo-responsive surfaces can switch their wettability (from wettable to non-wettable) and adhesion (from sticky to non-sticky) according to external temp. changes. These smart surfaces with switchable interfacial properties are playing increasingly important roles in a diverse range of biomedical applications; these controlling cell-adhesion behavior has shown great potential for tissue engineering and disease diagnostics. Herein we reviewed the recent progress of research on PNIPAAm-based thermo-responsive surfaces that can dynamically control cell adhesion behavior. The underlying response mechanisms and influencing factors for PNIPAAm-based surfaces to control cell adhesion are described first. Then, PNIPAAm-modified two-dimensional flat surfaces for cell-sheet engineering and PNIPAAm-modified three-dimensional nanostructured surfaces for diagnostics are summarized. We also provide a future perspective for the development of stimuli-responsive surfaces.
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      Rodriguez-Emmenegger, C.; Brynda, E.; Riedel, T.; Houska, M.; Šubr, V.; Alles, A. B.; Hasan, E.; Gautrot, J. E.; Huck, W. T. S. Polymer Brushes Showing Non-Fouling in Blood Plasma Challenge the Currently Accepted Design of Protein Resistant Surfaces. Macromol. Rapid Commun. 2011, 32, 952– 957,  DOI: 10.1002/marc.201100189
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      Polymer Brushes Showing Non-Fouling in Blood Plasma Challenge the Currently Accepted Design of Protein Resistant Surfaces
      Rodriguez-Emmenegger, Cesar; Brynda, Eduard; Riedel, Tomas; Houska, Milan; Subr, Vladimir; Alles, Aldo Bologna; Hasan, Erol; Gautrot, Julien E.; Huck, Wilhelm T. S.
      Macromolecular Rapid Communications (2011), 32 (13), 952-957CODEN: MRCOE3; ISSN:1022-1336. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Ultra-low-fouling poly[N-(2-hydroxypropyl) methacrylamide] (poly(HPMA)) brushes have been synthesized for the first time. Similar to the so far only ultra-low-fouling surface, poly(carboxybetaine acrylamide), the level of blood plasma fouling was below the detection limit of surface plasmon resonance (SPR, 0.03 ng/cm-2) despite being a hydrogen bond donor and displaying a moderate wettability, thus challenging the currently accepted views for the design of antifouling properties. The antifouling properties were preserved even after two years of storage. To demonstrate the potential of poly(HPMA) brushes for the prepn. of bioactive ultra-low fouling surfaces a label-free SPR immunosensor for detection of G Streptococcus was prepd.
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      Baggerman, J.; Smulders, M. M. J.; Zuilhof, H. Romantic Surfaces: A Systematic Overview of Stable, Biospecific, and Antifouling Zwitterionic Surfaces. Langmuir 2019, 35, 1072– 1084,  DOI: 10.1021/acs.langmuir.8b03360
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      Romantic Surfaces: A Systematic Overview of Stable, Biospecific, and Antifouling Zwitterionic Surfaces
      Baggerman, Jacob; Smulders, Maarten M. J.; Zuilhof, Han
      Langmuir (2019), 35 (5), 1072-1084CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      A review. This feature Article focuses on recent advances in the bioconjugation of surface-bound zwitterionic polymers for biospecific antifouling surfaces. Various approaches for the functionalization of antifouling zwitterionic polymers are systematically investigated, such as chain-end and side-chain functionalization. Side-chain functionalization methods can be further classified as those that are achieved through homopolymn. of custom-synthesized zwitterionic monomers equipped with reactive groups, or those that are achieved via synthesis of random or block copolymers combining different monomers with antifouling functionality and others with reactive groups. Several of the pros and cons of these approaches are outlined and discussed. Finally, some perspective and future directions of research are presented toward long-term stable, generically repelling surfaces that strongly and specifically adhere to a single component in a complex mixt.
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      Wischerhoff, E.; Badi, N.; Lutz, J.-F.; Laschewsky, A. Smart Bioactive Surfaces. Soft Matter 2010, 6, 705– 713,  DOI: 10.1039/B913594D
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      Smart bioactive surfaces
      Wischerhoff, Erik; Badi, Nezha; Lutz, Jean-Francois; Laschewsky, Andre
      Soft Matter (2010), 6 (4), 705-713CODEN: SMOABF; ISSN:1744-683X. (Royal Society of Chemistry)
      A review. The purpose of this highlight is to define the emerging field of bioactive surfaces. In recent years, various types of synthetic materials capable of "communicating" with biol. objects such as nucleic acids, proteins, polysaccharides, viruses, bacteria or living cells were described in the literature. This novel area of research certainly goes beyond the traditional field of smart materials and includes different types of sophisticated interactions with biol. entities, such as reversible adhesion, conformational control, biol.-triggered release and selective permeation. These novel materials may be 2D planar surfaces as well as colloidal objects or 3D scaffolds. Overall, they show great promise for numerous applications in biosciences and biotechnol. For instance, practical applications of bioactive surfaces in the fields of biosepn., cell engineering, biochips and stem-cell differentiation are briefly discussed herein.
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      Vorobii, M.; de los Santos Pereira, A.; Pop-Georgievski, O.; Kostina, N. Y.; Rodriguez-Emmenegger, C.; Percec, V. Synthesis of Non-fouling poly[N-(2-hydroxypropyl)methacrylamide] Brushes by Photoinduced SET-LRP. Polym. Chem. 2015, 6, 4210– 4220,  DOI: 10.1039/C5PY00506J
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      Synthesis of non-fouling poly[N-(2-hydroxypropyl)methacrylamide] brushes by photoinduced SET-LRP
      Vorobii, Mariia; de los Santos Pereira, Andres; Pop-Georgievski, Ognen; Kostina, Nina Yu.; Rodriguez-Emmenegger, Cesar; Percec, Virgil
      Polymer Chemistry (2015), 6 (23), 4210-4220CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
      Surface-initiated photoinduced single-electron transfer living radical polymn. (SET-LRP) was employed to assemble brushes of poly[N-(2-hydroxypropyl) methacrylamide] (poly(HPMA)) from Si surfaces. The linear increase in thickness of the poly(HPMA) brushes with time and the ability to prep. block copolymers indicate the living nature of this grafting-from process. Cu concns. ≥80 ppb were sufficient for this surface-initiated SET-LRP. Micropatterns of poly(HPMA) brushes on the Si surface were constructed for the first time by this method. Negligible fouling was obsd. after contact with undiluted blood plasma. This report provides the first example of nonfouling polymer brushes prepd. by SET-LRP of HPMA.
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      Jiang, S.; Cao, Z. Ultralow-Fouling, Functionalizable, and Hydrolyzable Zwitterionic Materials and Their Derivatives for Biological Applications. Adv. Mater. 2010, 22, 920– 932,  DOI: 10.1002/adma.200901407
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      Ultralow-Fouling, Functionalizable, and Hydrolyzable Zwitterionic Materials and Their Derivatives for Biological Applications
      Jiang, Shaoyi; Cao, Zhiqiang
      Advanced Materials (Weinheim, Germany) (2010), 22 (9), 920-932CODEN: ADVMEW; ISSN:0935-9648. (Wiley-VCH Verlag GmbH & Co. KGaA)
      A review. In recent years, zwitterionic materials such as poly(carboxybetaine) (pCB) and poly(sulfobetaine) (pSB) have been applied to a broad range of biomedical and engineering materials. Due to electrostatically induced hydration, surfaces coated with zwitterionic groups are highly resistant to nonspecific protein adsorption, bacterial adhesion, and biofilm formation. Among zwitterionic materials, pCB is unique due to its abundant functional groups for the convenient immobilization of biomols. pCB can also be prepd. in a hydrolyzable form as cationic pCB esters, which can kill bacteria or condense DNA. The hydrolysis of cationic pCB esters into nonfouling zwitterionic groups will lead to the release of killed microbes or the irreversible unpackaging of DNA. Furthermore, mixed-charge materials have been shown to be equiv. to zwitterionic materials in resisting nonspecific protein adsorption when they are uniformly mixed at the mol. scale.
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      Pereira, A. d. l. S.; Rodriguez-Emmenegger, C.; Surman, F.; Riedel, T.; Alles, A. B.; Brynda, E. Use of Pooled Blood Plasmas in the Assessment of Fouling Resistance. RSC Adv. 2014, 4, 2318– 2321,  DOI: 10.1039/C3RA43093F
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      Use of pooled blood plasmas in the assessment of fouling resistance
      Pereira, Andres de los Santos; Rodriguez-Emmenegger, Cesar; Surman, Frantisek; Riedel, Tomas; Alles, Aldo Bologna; Brynda, Eduard
      RSC Advances (2014), 4 (5), 2318-2321CODEN: RSCACL; ISSN:2046-2069. (Royal Society of Chemistry)
      The ability of a surface to resist fouling from blood plasma cannot be realistically estd. without measuring adsorption from real samples directly. Due to the variability of biol. samples, pooled blood plasma is normally used. We show that even when using pooled plasma, a comparison of antifouling surfaces strongly depends on its source.
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      Teunissen, L. W.; Kuzmyn, A. R.; Ruggeri, F. S.; Smulders, M. M. J.; Zuilhof, H. Thermoresponsive, Pyrrolidone-Based Antifouling Polymer Brushes. Adv. Mater. Interfaces 2022, 9, 2101717  DOI: 10.1002/admi.202101717
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      Thermoresponsive, pyrrolidone-based antifouling polymer brushes
      Teunissen, Lucas W.; Kuzmyn, Andriy R.; Ruggeri, Francesco S.; Smulders, Maarten M. J.; Zuilhof, Han
      Advanced Materials Interfaces (2022), 9 (6), 2101717CODEN: AMIDD2; ISSN:2196-7350. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Commonly, modification of surfaces with thermoresponsive polymers is performed using poly(N-isopropylacrylamide) (poly(NIPAM)). However, integration of poly(NIPAM) with a second polymer to obtain more complex copolymer structures has proven challenging due to inherently poorly controllable polymn. characteristics of acrylamides. In this study, N-(2-methacryloyloxyethyl)pyrrolidone (NMEP) is synthesized and polymd. under controlled conditions from silicon oxide substrates via surface-initiated atom transfer radical polymn. (SI-ATRP) to produce thermoresponsive poly(NMEP) brushes. The livingness of the brushes is demonstrated by reinitiation of poly(NMEP) brushes using oligo(ethylene glycol) Me ether methacrylate to obtain diblock copolymer brushes. Following extensive characterization, the reversible thermoresponsive behavior of these poly(NMEP) brushes is demonstrated using phase-controlled AFM topog. measurements in an aq. liq. environment. These measurements indicate that at 27°C the poly(NMEP) brushes are solvated and extend away from the surface. Finally, to investigate the potential applicability of poly(NMEP) brushes in biomedical devices, the antifouling properties of the coating are tested in aq. media contg. BSA, fibrinogen, or 10% dild. human serum using quartz crystal microbalance with dissipation monitoring (QCM-D). These measurements reveal very good antifouling properties, even when exposed to 10% dild. human serum.
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    38. 38
      van der Weg, K. J.; Ritsema van Eck, G. C.; de Beer, S. Polymer Brush Friction in Cylindrical Geometries. Lubricants 2019, 7, 84  DOI: 10.3390/lubricants7100084
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      Yu, Y.; Brió Pérez, M.; Cao, C.; de Beer, S. Switching (Bio-) adhesion and Friction in Liquid by Stimulus Responsive Polymer Coatings. Eur. Polym. J. 2021, 147, 110298  DOI: 10.1016/j.eurpolymj.2021.110298
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      Switching (bio-) adhesion and friction in liquid by stimulus responsive polymer coatings
      Yu, Yunlong; Brio Perez, Maria; Cao, Cong; de Beer, Sissi
      European Polymer Journal (2021), 147 (), 110298CODEN: EUPJAG; ISSN:0014-3057. (Elsevier Ltd.)
      A review. Controlling (bio-) adhesion and friction is essential in many bio-medical and industrial applications. Such control can be achieved using stimulus responsive polymers. Coatings composed of these polymers are able to adapt their physicochem. properties to changes in the surrounding environment, such as temp., light, pH, electronic and magnetic fields. This responsive behavior can be employed in the development of advanced technologies as for example underwater adhesives, self-adhering wound dressings, soft robotics and targeted drug delivery. In this review, we present an overview of the current status of research on controlling adhesion and friction in liq. environments using surface-bound stimulus responsive polymers. We focus in particular on polymer brushes and the role of specific interactions between the surfaces in the contact. These interactions can give rise to complex relations between swelling of the polymer film and its tribomech. properties. A careful characterization of the change in specific interactions upon varying the environment is, therefore, crit. for the design of effective smart coatings. We will finish this review with an overview of the open questions in the field and a discussion of the potential impact that the polymer-based smart adhesives and lubricants can have on industry and society.
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    40. 40
      Yeole, N. Thiocarbonylthio Compounds. Synlett 2010, 2010, 1572– 1573,  DOI: 10.1055/s-0029-1219938
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      Moad, G.; Rizzardo, E.; Thang, S. H. Radical addition–fragmentation chemistry in polymer synthesis. Polymer 2008, 49, 1079– 1131,  DOI: 10.1016/j.polymer.2007.11.020
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      Radical addition-fragmentation chemistry in polymer synthesis
      Moad, Graeme; Rizzardo, Ezio; Thang, San H.
      Polymer (2008), 49 (5), 1079-1131CODEN: POLMAG; ISSN:0032-3861. (Elsevier Ltd.)
      A review on the development of addn.-fragmentation chain transfer agents and related ring-opening monomers highlighting recent innovation in these areas. The major part of this review deals with reagents that give reversible addn.-fragmentation chain transfer (RAFT). These reagents include dithioesters, trithiocarbonates, dithiocarbamates and xanthates. The RAFT process is a versatile method for conferring living characteristics on radical polymns. providing unprecedented control over mol. wt., mol. wt. distribution, compn. and architecture. It is suitable for most monomers polymerizable by radical polymn. and is robust under a wide range of reaction conditions. It provides a route to functional polymers, cyclopolymers, gradient copolymers, block polymers and star polymers.
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    42. 42
      Niu, J.; Page, Z. A.; Dolinski, N. D.; Anastasaki, A.; Hsueh, A. T.; Soh, H. T.; Hawker, C. J. Rapid Visible Light-Mediated Controlled Aqueous Polymerization with In Situ Monitoring. ACS Macro Lett. 2017, 6, 1109– 1113,  DOI: 10.1021/acsmacrolett.7b00587
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      Rapid Visible Light-Mediated Controlled Aqueous Polymerization with In Situ Monitoring
      Niu, Jia; Page, Zachariah A.; Dolinski, Neil D.; Anastasaki, Athina; Hsueh, Andy T.; Soh, H. Tom; Hawker, Craig J.
      ACS Macro Letters (2017), 6 (10), 1109-1113CODEN: AMLCCD; ISSN:2161-1653. (American Chemical Society)
      We report a simple procedure for rapid, visible light-mediated, controlled radical polymn. in aq. solns. Based on the photoelectron transfer reversible addn.-fragmentation chain transfer (PET-RAFT) polymn., fast chain propagation at room temp. in water was achieved in the presence of reductant and without prior deoxygenation. A systematic study correlating irradn. intensity and polymn. kinetics, enabled by in situ NMR spectroscopy, provided optimized reaction conditions. The versatility of this procedure was demonstrated through a rapid triblock copolymer synthesis, and incorporation of water-labile activated esters for direct conjugation of hydrophilic small mols. and proteins. In addn., this technique boasts excellent temporal control and provides a wide range of macromol. materials with controlled mol. wts. and narrow mol. wt. distributions.
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    43. 43
      Lueckerath, T.; Strauch, T.; Koynov, K.; Barner-Kowollik, C.; Ng, D. Y. W.; Weil, T. DNA–Polymer Conjugates by Photoinduced RAFT Polymerization. Biomacromolecules 2019, 20, 212– 221,  DOI: 10.1021/acs.biomac.8b01328
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      DNA-Polymer Conjugates by Photoinduced RAFT Polymerization
      Lueckerath, Thorsten; Strauch, Tina; Koynov, Kaloian; Barner-Kowollik, Christopher; Ng, David Y. W.; Weil, Tanja
      Biomacromolecules (2019), 20 (1), 212-221CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)
      Conventional grafting-to approaches to DNA-polymer conjugates are often limited by low reaction yields due to the sterically hindered coupling of a presynthesized polymer to DNA. The grafting-from strategy, in contrast, allows one to directly graft polymers from an initiator that is covalently attached to DNA. Herein, we report blue-light-mediated reversible addn.-fragmentation chain-transfer (Photo-RAFT) polymn. from two different RAFT agent-terminated DNA sequences using Eosin Y as the photocatalyst in combination with ascorbic acid. Three monomer families (methacrylates, acrylates and acrylamides) were successfully polymd. from DNA employing Photo-RAFT polymn. We demonstrate that the length of the grown polymer chain can be varied by altering the monomer to DNA-initiator ratio, while the self-assembly features of the DNA strands were maintained. In summary, we describe a convenient, light-mediated approach toward DNA-polymer conjugates via the grafting-from approach.
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    44. 44
      Niu, J.; Lunn, D. J.; Pusuluri, A.; Yoo, J. I.; O’Malley, M. A.; Mitragotri, S.; Soh, H. T.; Hawker, C. J. Engineering Live Cell Surfaces with Functional Polymers via Cytocompatible Controlled Radical Polymerization. Nat. Chem. 2017, 9, 537– 545,  DOI: 10.1038/nchem.2713
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      Engineering live cell surfaces with functional polymers via cytocompatible controlled radical polymerization
      Niu, Jia; Lunn, David J.; Pusuluri, Anusha; Yoo, Justin I.; O'Malley, Michelle A.; Mitragotri, Samir; Soh, H. Tom; Hawker, Craig J.
      Nature Chemistry (2017), 9 (6), 537-545CODEN: NCAHBB; ISSN:1755-4330. (Nature Publishing Group)
      The capability to graft synthetic polymers onto the surfaces of live cells offers the potential to manipulate and control their phenotype and underlying cellular processes. Conventional grafting-to strategies for conjugating preformed polymers to cell surfaces are limited by low polymer grafting efficiency. Here we report an alternative grafting-from strategy for directly engineering the surfaces of live yeast and mammalian cells through cell surface-initiated controlled radical polymn. By developing cytocompatible PET-RAFT (photoinduced electron transfer-reversible addn.-fragmentation chain-transfer polymn.), synthetic polymers with narrow polydispersity (Mw/Mn < 1.3) could be obtained at room temp. in 5 min. This polymn. strategy enables chain growth to be initiated directly from chain-transfer agents anchored on the surface of live cells using either covalent attachment or non-covalent insertion, while maintaining high cell viability. Compared with conventional grafting-to approaches, these methods significantly improve the efficiency of grafting polymer chains and enable the active manipulation of cellular phenotypes.
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    45. 45
      Li, M.; Fromel, M.; Ranaweera, D.; Rocha, S.; Boyer, C.; Pester, C. W. SI-PET-RAFT: Surface-Initiated Photoinduced Electron Transfer-Reversible Addition–Fragmentation Chain Transfer Polymerization. ACS Macro Lett. 2019, 8, 374– 380,  DOI: 10.1021/acsmacrolett.9b00089
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      45
      SI-PET-RAFT: Surface-Initiated Photoinduced Electron Transfer-Reversible Addition-Fragmentation Chain Transfer Polymerization
      Li, Mingxiao; Fromel, Michele; Ranaweera, Dhanesh; Rocha, Sergio; Boyer, Cyrille; Pester, Christian W.
      ACS Macro Letters (2019), 8 (4), 374-380CODEN: AMLCCD; ISSN:2161-1653. (American Chemical Society)
      In this communication, surface-initiated photoinduced electron transfer-reversible addn.-fragmentation chain transfer polymn. (SI-PET-RAFT) is introduced. SI-PET-RAFT affords functionalization of surfaces with spatiotemporal control and provides oxygen tolerance under ambient conditions. All hallmarks of controlled radical polymn. (CRP) are met, affording well-defined polymn. kinetics, and chain end retention to allow subsequent extension of active chain ends to form block copolymers. The modularity and versatility of SI-PET-RAFT is highlighted through significant flexibility with respect to the choice of monomer, light source and wavelength, and photoredox catalyst. The ability to obtain complex patterns in the presence of air is a significant contribution to help pave the way for CRP-based surface functionalization into com. application.
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    46. 46
      Kuzmyn, A. R.; Teunissen, L. W.; Fritz, P.; van Lagen, B.; Smulders, M. M. J.; Zuilhof, H. Diblock and Random Antifouling Bioactive Polymer Brushes on Gold Surfaces by Visible-Light-Induced Polymerization (SI-PET-RAFT) in Water. Adv. Mater. Interfaces 2022, 9, 2101784  DOI: 10.1002/admi.202101784
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      46
      Diblock and random antifouling bioactive polymer brushes on gold surfaces by visible-light-induced polymerization in water
      Kuzmyn, Andriy R.; Teunissen, Lucas W.; Fritz, Pina; van Lagen, Barend; Smulders, Maarten M. J.; Zuilhof, Han
      Advanced Materials Interfaces (2022), 9 (3), 2101784CODEN: AMIDD2; ISSN:2196-7350. (Wiley-VCH Verlag GmbH & Co. KGaA)
      Surface-initiated photoinduced electron-transfer-reversible addn.-fragmentation chain transfer (SI-PET-RAFT) is, for the first time, used for the creation of antifouling polymer brushes on gold surfaces based on three monomers: oligo(ethylene glycol) Me ether methacrylate (MeOEGMA), N-(2-hydroxypropyl) methacrylamide (HPMA), and carboxybetaine methacrylamide (CBMA). These coatings are subsequently characterized by XPS and ellipsometry. The living nature of this polymn. allows for the creation of random and diblock copolymer brushes, which are based on HPMA (superb antifouling) and CBMA (good antifouling and functionalizable via activated ester chem.). The polymer brushes demonstrate good antifouling properties against undiluted human serum, as monitored by quartz crystal microbalance with dissipation (QCM-D) and surface plasmon resonance (SPR) spectroscopy in real time. The amt. of immobilization of bioactive moieties, here an antibody immobilized using N-succinimidyl ester-1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (NHS-EDC) coupling, in the diblock and random copolymer brushes is monitored by SPR, and is analyzed with respect to the brush structure, and is shown to be superior in the diblock copolymer brush. This approach represents a scalable, robust, mild, oxygen-tolerant, and heavy-metal-free route toward the prodn. of antifouling and functional copolymer brushes (on gold surfaces) that open up applications in biosensing and tissue engineering.
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    47. 47
      Roeven, E.; Kuzmyn, A. R.; Scheres, L.; Baggerman, J.; Smulders, M. M. J.; Zuilhof, H. PLL–Poly(HPMA) Bottlebrush-Based Antifouling Coatings: Three Grafting Routes. Langmuir 2020, 36, 10187– 10199,  DOI: 10.1021/acs.langmuir.0c01675
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      47
      PLL-Poly(HPMA) Bottlebrush-Based Antifouling Coatings: Three Grafting Routes
      Roeven, Esther; Kuzmyn, Andriy R.; Scheres, Luc; Baggerman, Jacob; Smulders, Maarten M. J.; Zuilhof, Han
      Langmuir (2020), 36 (34), 10187-10199CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      In this work, we compare three routes to prep. antifouling coatings that consist of poly(L-lysine)-poly(N-(2-hydroxypropyl)methacrylamide) bottlebrushes. The poly(L-lysine) (PLL) backbone is self-assembled onto the surface by charged-based interactions between the lysine groups and the neg. charged silicon oxide surface, whereas the poly(N-(2-hydroxypropyl)methacrylamide) [poly(HPMA)] side chains, grown by reversible addn.-fragmentation chain-transfer (RAFT) polymn., provide antifouling properties to the surface. First, the PLL-poly(HPMA) coatings are synthesized in a bottom-up fashion through a grafting-from approach. In this route, the PLL is self-assembled onto a surface, after which a polymn. agent is immobilized, and finally HPMA is polymd. from the surface. In the second explored route, the PLL is modified in soln. by a RAFT agent to create a macroinitiator. After self-assembly of this macroinitiator onto the surface, poly(HPMA) is polymd. from the surface by RAFT. In the third and last route, the whole PLL-poly(HPMA) bottlebrush is initially synthesized in soln. To this end, HPMA is polymd. from the macroinitiator in soln. and the PLL-poly(HPMA) bottlebrush is then self-assembled onto the surface in just one step (grafting-to approach). Addnl., in this third route, we also design and synthesize a bottlebrush polymer with a PLL backbone and poly(HPMA) side chains, with the latter contg. 5% carboxybetaine (CB) monomers that eventually allow for addnl. (bio)functionalization in soln. or after surface immobilization. These three routes are evaluated in terms of ease of synthesis, scalability, ease of characterization, and a preliminary investigation of their antifouling performance. All three coating procedures result in coatings that show antifouling properties in single-protein antifouling tests. This method thus presents a new, simple, versatile, and highly scalable approach for the manufg. of PLL-based bottlebrush coatings that can be synthesized partly or completely on the surface or in soln., depending on the desired prodn. process and/or application.
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    48. 48
      Xu, J.; Shanmugam, S.; Duong, H. T.; Boyer, C. Organo-photocatalysts for Photoinduced Electron Transfer-reversible Addition–fragmentation Chain Transfer (PET-RAFT) Polymerization. Polym. Chem. 2015, 6, 5615– 5624,  DOI: 10.1039/C4PY01317D
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      48
      Organo-photocatalysts for photoinduced electron transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization
      Xu, Jiangtao; Shanmugam, Sivaprakash; Duong, Hien T.; Boyer, Cyrille
      Polymer Chemistry (2015), 6 (31), 5615-5624CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)
      In this article, we investigated a series of organo-dyes, including methylene blue, fluorescein, Rhodamine 6G, Nile red and eosin Y, to perform a visible light-mediated controlled/"living" radical polymn. of methacrylates. We demonstrate that eosin Y and fluorescein were efficient catalysts to activate a photoinduced electron transfer-reversible addn.-fragmentation chain transfer (PET-RAFT) mechanism. The concn. of eosin Y was varied from 10 to 100 ppm with respect to monomers. This polymn. technique yielded well-defined (co)polymers with a good control of the mol. wts. ranging from 10 000 to 100 000 g mol-1 and low polydispersities (PDI < 1.30). A variety of functional monomers, including N,N-dimethylaminoethyl methacrylate, hydroxyl Et methacrylate, pentafluorophenyl methacrylate, glycidyl methacrylate, oligo(ethylene glycol) Me ether methacrylate (OEGMA), and methacrylic acid, were successfully polymd. Finally, the addn. of a tertiary amine, such as triethylamine, afforded the polymn. in the presence of air via a reductive quenching cycle. Different diblock polymethacrylate copolymers, i.e.PMMA-b-POEGMA and PMMA-b-PMMA, were prepd. to demonstrate the high end group fidelity.
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    49. 49
      Cheng, G.; Li, G.; Xue, H.; Chen, S.; Bryers, J. D.; Jiang, S. Zwitterionic Carboxybetaine Polymer Surfaces and their Resistance to Long-term Biofilm Formation. Biomaterials 2009, 30, 5234– 5240,  DOI: 10.1016/j.biomaterials.2009.05.058
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      Zwitterionic carboxybetaine polymer surfaces and their resistance to long-term biofilm formation
      Cheng, Gang; Li, Guozhu; Xue, Hong; Chen, Shengfu; Bryers, James D.; Jiang, Shaoyi
      Biomaterials (2009), 30 (28), 5234-5240CODEN: BIMADU; ISSN:0142-9612. (Elsevier Ltd.)
      The authors report a systematic study of zwitterionic poly(carboxybetaine methacrylate) (pCBMA) grafted from glass surfaces via atom transfer radical polymn. (ATRP) for their resistance to long-term bacterial biofilm formation. Results show that pCBMA-grafted surfaces are highly resistant to non-specific protein adsorption (fibrinogen and undiluted blood plasma) at 25, 30 and 37°. Long-term (over 24 h) colonization of two bacterial strains (Pseudomonas aeruginosa PAO1 and Pseudomonas putida strain 239) on pCBMA surface was studied using a parallel flow cell at 25, 30 and 37°. Uncoated glass cover slips were chosen as the pos. ref. Results show that pCBMA coatings reduced long-term biofilm formation of P. aeruginosa up to 240 h by 95% at 25° and for 64 h by 93% at 37°, and suppressed P. putida biofilm accumulation up to 192 h by 95% at 30°, with respect to the glass ref. The ability of pCBMA coatings to resist non-specific protein adsorption and significantly retard bacterial biofilm formation makes it a very promising material for biomedical and industrial applications.
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    50. 50
      van Andel, E.; Lange, S. C.; Pujari, S. P.; Tijhaar, E. J.; Smulders, M. M. J.; Savelkoul, H. F. J.; Zuilhof, H. Systematic Comparison of Zwitterionic and Non-Zwitterionic Antifouling Polymer Brushes on a Bead-Based Platform. Langmuir 2019, 35, 1181– 1191,  DOI: 10.1021/acs.langmuir.8b01832
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      Systematic Comparison of Zwitterionic and Non-Zwitterionic Antifouling Polymer Brushes on a Bead-Based Platform
      van Andel, Esther; Lange, Stefanie C.; Pujari, Sidharam P.; Tijhaar, Edwin J.; Smulders, Maarten M. J.; Savelkoul, Huub F. J.; Zuilhof, Han
      Langmuir (2019), 35 (5), 1181-1191CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      Nonspecific adsorption of biomols. to solid surfaces, a process called biofouling, is a major concern in many biomedical applications. Great effort has been made in the development of antifouling polymer coatings that are capable of repelling the nonspecific adsorption of proteins, cells, and micro-organisms. In this respect, we herein contribute to understanding the factors that det. which polymer brush results in the best antifouling coating. To this end, we compared five different monomers: two sulfobetaines, a carboxybetaine, a phosphocholine, and a hydroxyl acrylamide. The antifouling coatings were analyzed using our previously described bead-based method with flow cytometry as the read-out system. This method allows for the quick and automated anal. of thousands of beads per s, enabling fast anal. and good statistics. We report the first direct comparison made between a sulfobetaine with opposite charges sepd. by two and three methylene groups and a carboxybetaine bearing two sepg. methylene groups. It was concluded that both the distance between opposite charges and the nature of the anionic groups have a distinct effect on the antifouling performance. Phosphocholines and simple hydroxyl acrylamides are not often compared with the betaines. However, here we found that they perform equally well or even better, yielding the following overall antifouling ranking: HPMAA ≥ PCMA-2 ≈ CBMAA-2 > SBMAA-2 > SBMAA-3 » nonmodified beads (HPMAA being the best).
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      Sah, P.; Alfaro-Murillo, J. A.; Fitzpatrick, M. C.; Neuzil, K. M.; Meyers, L. A.; Singer, B. H.; Galvani, A. P. Future Epidemiological and Economic Impacts of Universal Influenza Vaccines. Proc. Natl. Acad. Sci. U.S.A. 2019, 116, 20786– 20792,  DOI: 10.1073/pnas.1909613116
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      Future epidemiological and economic impacts of universal influenza vaccines
      Sah, Pratha; Alfaro-Murillo, Jorge A.; Fitzpatrick, Meagan C.; Neuzil, Kathleen M.; Meyers, Lauren A.; Singer, Burton H.; Galvani, Alison P.
      Proceedings of the National Academy of Sciences of the United States of America (2019), 116 (41), 20786-20792CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
      The efficacy of influenza vaccines, currently at 44%, is limited by the rapid antigenic evolution of the virus and a manufg. process that can lead to vaccine mismatch. The National Institute of Allergy and Infectious Diseases (NIAID) recently identified the development of a universal influenza vaccine with an efficacy of at least 75% as a high scientific priority. The US Congress approved $130 million funding for the 2019 fiscal year to support the development of a universal vaccine, and another $1 billion over 5 y has been proposed in the Flu Vaccine Act. Using a model of influenza transmission, we evaluated the population-level impacts of universal influenza vaccines distributed according to empirical age-specific coverage at multiple scales in the United States. We est. that replacing just 10% of typical seasonal vaccines with 75% efficacious universal vaccines would avert ∼5.3 million cases, 81,000 hospitalizations, and 6,300 influenza-related deaths per yr. This would prevent over $1.1 billion in direct health care costs compared to a typical season, based on av. data from the 2010-11 to 2018-19 seasons. A complete replacement of seasonal vaccines with universal vaccines is projected to prevent 17 million cases, 251,000 hospitalizations, 19,500 deaths, and $3.5 billion in direct health care costs. States with high per-hospitalization medical expenses along with a large proportion of elderly residents are expected to receive the max. economic benefit. Replacing even a fraction of seasonal vaccines with universal vaccines justifies the substantial cost of vaccine development.
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    52. 52
      Vaisocherová, H.; Ševců, V.; Adam, P.; Špačková, B.; Hegnerová, K.; de los Santos Pereira, A.; Rodriguez-Emmenegger, C.; Riedel, T.; Houska, M.; Brynda, E.; Homola, J. Functionalized Ultra-low Fouling Carboxy- and Hydroxy-functional Surface Platforms: Functionalization Capacity, Biorecognition Capability and Resistance to Fouling from Undiluted Biological Media. Biosens. Bioelectron. 2014, 51, 150– 157,  DOI: 10.1016/j.bios.2013.07.015
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      Functionalized ultra-low fouling carboxy- and hydroxy-functional surface platforms: functionalization capacity, biorecognition capability and resistance to fouling from undiluted biological media
      Vaisocherova, Hana; Sevcu, Veronika; Adam, Pavel; Spackova, Barbora; Hegnerova, Katerina; de los Santos Pereira, Andres; Rodriguez-Emmenegger, Cesar; Riedel, Tomas; Houska, Milan; Brynda, Eduard; Homola, Jiri
      Biosensors & Bioelectronics (2014), 51 (), 150-157CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
      The non-specific binding of non-target species to functionalized surfaces of biosensors continues to be challenge for biosensing in real-world media. Three different low-fouling and functionalizable surface platforms were employed to study the effect of functionalization on fouling resistance from several types of undiluted media including blood plasma and food media. The surface platforms investigated in this work included two polymer brushes: hydroxy-functional poly(2-hydroxyethyl methacrylate) (pHEMA) and carboxy-functional poly(carboxybetaine acrylamide) (pCBAA), and a std. OEG-based carboxy-functional alkanethiolate self-assembled monolayer (AT-SAM). The wet and dry polymer brushes were analyzed by AFM, ellipsometry, FT-IRRAS, and surface plasmon resonance (SPR). The surfaces were functionalized by the covalent attachment of antibodies, streptavidin, and oligonucleotides and the binding and biorecognition characteristics of the coatings were compared. We found that functionalization did not substantially affect the ultra-low fouling properties of pCBAA (plasma fouling of ∼20 ng/cm2), a finding in contrast with pHEMA that completely lost its resistance to fouling after the activation of hydroxyl groups. Blocking a functionalized AT-SAM covalently with BSA decreased fouling down to the level comparable to unblocked pCBAA. However, the biorecognition capability of blocked functionalized AT-SAM was poor in comparison with functionalized pCBAA. Limits of detection of Escherichia coli O157:H7 in undiluted milk were detd. to be 6×104, 8×105, and 6×105 cells/mL for pCBAA, pHEMA, and AT-SAM-blocked, resp. Effect of analyte size on biorecognition activity of functionalized coatings was investigated and it was shown that the best performance in terms of overall fouling resistance and biorecognition capability is provided by pCBAA.
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    53. 53
      Vaisocherová-Lísalová, H.; Surman, F.; Víšová, I.; Vala, M.; Špringer, T.; Ermini, M. L.; Šípová, H.; Šedivák, P.; Houska, M.; Riedel, T.; Pop-Georgievski, O.; Brynda, E.; Homola, J. Copolymer Brush-Based Ultralow-Fouling Biorecognition Surface Platform for Food Safety. Anal. Chem. 2016, 88, 10533– 10539,  DOI: 10.1021/acs.analchem.6b02617
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      Copolymer Brush-Based Ultralow-Fouling Biorecognition Surface Platform for Food Safety
      Vaisocherova-Lisalova, Hana; Surman, Frantisek; Visova, Ivana; Vala, Milan; Springer, Tomas; Ermini, Maria Laura; Sipova, Hana; Sedivak, Petr; Houska, Milan; Riedel, Tomas; Pop-Georgievski, Ognen; Brynda, Eduard; Homola, Jiri
      Analytical Chemistry (Washington, DC, United States) (2016), 88 (21), 10533-10539CODEN: ANCHAM; ISSN:0003-2700. (American Chemical Society)
      Functional polymer coatings that combine the ability to resist non-specific fouling from complex media with high biorecognition element (BRE) immobilization capacity represent an emerging class of new functional materials for a no. of bioanal. and biosensor technologies for medical diagnostics, security, and food safety. Here we report on a random copolymer brush surface - poly(CBMAA-ran-HPMAA) - providing high BRE immobilization capacity while simultaneously exhibiting ultralow-fouling behavior in complex food media. We demonstrate that both the functionalization and fouling resistance capabilities of such copolymer brushes can be tuned by changing the surface contents of the two monomer units: non-ionic N-(2-hydroxypropyl) methacrylamide (HPMAA) and carboxy-functional zwitterionic carboxybetaine methacrylamide (CBMAA). It is demonstrated that the resistance to fouling decreases with the surface content of CBMAA; poly(CBMAA-ran-HPMAA) brushes with CBMAA molar content up to 15 mol% maintain excellent resistance to fouling from a variety of homogenized foods (hamburger, cucumber, milk, and lettuce) even after covalent attachment of BREs to carboxy-groups of CBMAA. The poly(CBMAA 15 mol%-ran-HPMAA) brushes functionalized with antibodies are demonstrated to exhibit fouling resistance from food samples by up to three orders of magnitude better when compared with the widely used low-fouling carboxy-functional oligo(ethylene glycol) (OEG)-based alkanethiolate self-assembled monolayers (AT SAMs) and furthermore, by up to two orders of magnitude better when compared with the most successful ultralow-fouling biorecognition coatings - poly(carboxybetaine acrylamide), poly(CBAA). Using model SPR detections of foodborne bacterial pathogens in homogenized foods it is also demonstrated that the antibody-functionalized poly(CBMAA 15 mol%-ran-HPMAA) brush exhibits superior biorecognition properties over the poly(CBAA).
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      Chaibi, W.; Ziane, A.; Benzehaim, Z.; Bennabi, L.; Guemra, K. Synthesis and Characterization of Cationic Poly(N-[3-Hexyldimethyl-Aminopropyl] Methacrylamide Bromide) Water-Soluble Polymer. Mater. Sci. Appl. Chem. 2016, 33, 40– 44,  DOI: 10.1515/msac-2016-0008
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      Easton, C. D.; Kinnear, C.; McArthur, S. L.; Gengenbach, T. R. Practical Guides for x-ray Photoelectron Spectroscopy: Analysis of polymers. J. Vac. Sci. Technol., A 2020, 38, 023207  DOI: 10.1116/1.5140587
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      Practical guides for x-ray photoelectron spectroscopy: Analysis of polymers
      Easton, Christopher D.; Kinnear, Calum; McArthur, Sally L.; Gengenbach, Thomas R.
      Journal of Vacuum Science & Technology, A: Vacuum, Surfaces, and Films (2020), 38 (2), 023207CODEN: JVTAD6; ISSN:0734-2101. (American Institute of Physics)
      A review. XPS is widely used to identify and quantify the elements present at the surface of polymeric materials. The energy distribution of photoelectrons emitted from these elements contains information about their chem. state, potentially allowing the analyst to identify and quantify specific functional groups. These functional groups may originate from the synthesis and processing of the polymers, from postsynthetic modifications such as surface grafting, or indeed may be unrelated to the polymer (additives and contaminants). Extg. reliable and meaningful information from XPS data is not trivial and relies on careful and appropriate experimentation, including exptl. design, sample prepn., data collection, data processing, and data interpretation. Here, the authors outline some of these challenges when performing XPS anal. of polymers and provide practical examples to follow. This guide will cover all relevant aspects over the course of a typical expt., including tips and considerations when designing the expt., sample prepn., charge neutralization, x-ray induced sample damage, depth profiling, data anal. and interpretation, and, finally, reporting of results. Many of these topics are more widely applicable to insulating org. materials, and the recommendations of this guide will help to ensure that data is collected and interpreted using current best practices. (c) 2020 American Institute of Physics.
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      Nečas, D.; Klapetek, P. Gwyddion: an Open-source Software for SPM Data Analysis. Open Phys. 2012, 10, 181– 188,  DOI: 10.2478/s11534-011-0096-2
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      Bergervoet, S. A.; Ho, C. K. Y.; Heutink, R.; Bossers, A.; Beerens, N. Spread of Highly Pathogenic Avian Influenza (HPAI) H5N5 Viruses in Europe in 2016–2017 Appears Related to the Timing of Reassortment Events. Viruses 2019, 11, 501– 517,  DOI: 10.3390/v11060501
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      Spread of highly pathogenic avian influenza (HPAI) H5N5 viruses in europe in 2016-2017 appears related to the timing of reassortment events
      Bergervoet, Saskia A.; Ho, Cynthia K. Y.; Heutink, Rene; Bossers, Alex; Beerens, Nancy
      Viruses (2019), 11 (6), 501CODEN: VIRUBR; ISSN:1999-4915. (MDPI AG)
      During the epizootic of highly pathogenic avian influenza (HPAI) H5N8 virus in Europe in 2016-2017, HPAI viruses of subtype H5N5 were also isolated. However, the detection of H5N5 viruses was limited compared to H5N8. In this study, we show that the genetic constellation of a newly isolated H5N5 virus is different from two genotypes previously identified in the Netherlands. The introduction and spread of the three H5N5 genotypes in Europe was studied using spatiotemporal and genetic anal. This demonstrated that the genotypes were isolated in distinguishable phases of the epizootic, and suggested multiple introductions of H5N5 viruses into Europe followed by local spread. We estd. the timing of the reassortment events, which suggested that the genotypes emerged after the start of autumn migration. This may have prevented large-scale spread of the H5N5 viruses on wild bird breeding sites before introduction into Europe. Expts. in primary chicken and duck cells revealed only minor differences in cytopathogenicity and replication kinetics between H5N5 genotypes and H5N8. These results suggest that the limited spread of HPAI H5N5 viruses is related to the timing of the reassortment events rather than changes in virus pathogenicity or replication kinetics.
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      Gerhards, N. M.; Cornelissen, J. B. W. J.; van Keulen, L. J. M.; Harders-Westerveen, J.; Vloet, R.; Smid, B.; Vastenhouw, S.; van Oort, S.; Hakze-van der Honing, R. W.; Gonzales, J. L.; Stockhofe-Zurwieden, N.; de Jong, R.; van der Poel, W. H. M.; Vreman, S.; Kortekaas, J.; Wichgers Schreur, P. J.; Oreshkova, N. Predictive Value of Precision-Cut Lung Slices for the Susceptibility of Three Animal Species for SARS-CoV-2 and Validation in a Refined Hamster Model. Pathogens 2021, 10, 824  DOI: 10.3390/pathogens10070824
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      Predictive Value of Precision-Cut Lung Slices for the Susceptibility of Three Animal Species for SARS-CoV-2 and Validation in a Refined Hamster Model
      Gerhards, Nora M.; Cornelissen, Jan B. W. J.; van Keulen, Lucien J. M.; Harders-Westerveen, Jose; Vloet, Rianka; Smid, Bregtje; Vastenhouw, Stephanie; van Oort, Sophie; Hakze-van der Honing, Renate W.; Gonzales, Jose L.; Stockhofe-Zurwieden, Norbert; de Jong, Rineke; van der Poel, Wim H. M.; Vreman, Sandra; Kortekaas, Jeroen; Wichgers Schreur, Paul J.; Oreshkova, Nadia
      Pathogens (2021), 10 (7), 824CODEN: PATHCD; ISSN:2076-0817. (MDPI AG)
      In assessing species susceptibility for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and in the search for an appropriate animal model, multiple research groups around the world inoculated a broad range of animal species using various SARS-CoV-2 strains, doses and administration routes. Although in silico analyses based on receptor binding and diverse in vitro cell cultures were valuable, exact prediction of species susceptibility based on these tools proved challenging. Here, we assessed whether precision-cut lung slices (PCLS) could facilitate the selection of animal models, thereby reducing animal experimentation. Pig, hamster and cat PCLS were incubated with SARS-CoV-2 and virus replication was followed over time. Virus replicated efficiently in PCLS from hamsters and cats, while no evidence of replication was obtained for pig PCLS. These data corroborate the findings of many research groups that have investigated the susceptibility of hamsters, pigs and cats towards infection with SARS-CoV-2. Our findings suggest that PCLS can be used as convenient tool for the screening of different animal species for sensitivity to newly emerged viruses. To validate our results obtained in PCLS, we employed the hamster model. Hamsters were inoculated with SARS-CoV-2 via the intranasal route. Susceptibility to infection was evaluated by body wt. loss, viral loads in oropharyngeal swabs and respiratory tissues and lung pathol. The broadly used hamster model was further refined by including activity tracking of the hamsters by an activity wheel as a very robust and sensitive parameter for clin. health. In addn., to facilitate the quantification of pathol. in the lungs, we devised a semi-quant. scoring system for evaluating the degree of histol. changes in the lungs. The inclusion of these addnl. parameters refined and enriched the hamster model, allowing for the generation of more data from a single expt.
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      Solubility of polymers
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      A review with 39 refs. covers the detn. of polymer soly. limits and crit. conditions and the general phenomena obsd. with systems contg. high mol. wt. materials.
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      Abraham, S.; Unsworth, L. D. Multi-functional Initiator and Poly(carboxybetaine methacrylamides) for Building biocompatible Surfaces Using “Nitroxide Mediated Free Radical Polymerization” Strategies. J. Polym. Sci., Part A: Polym. Chem. 2011, 49, 1051– 1060,  DOI: 10.1002/pola.24517
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      Multi-functional initiator and poly(carboxybetaine methacrylamides) for building biocompatible surfaces using "nitroxide mediated free radical polymerization" strategies
      Abraham, Sinoj; Unsworth, Larry D.
      Journal of Polymer Science, Part A: Polymer Chemistry (2011), 49 (5), 1051-1060CODEN: JPACEC; ISSN:0887-624X. (John Wiley & Sons, Inc.)
      Biomaterials generally suffer from rapid nonspecific protein adsorption, which initiates many deleterious host responses, and complex chemistries that are employed to facilitate cellular interactions. A chem. approach that, based upon current literature, combines a nonfouling architecture with a biomemtic cell-adhesive end-group, is presented. Namely, surface-initiated polymn. of zwitterionic [poly (carboxybetaine methacrylamide)] brushes, with controlled charge densities and phosphonate head groups. Nitroxide mediated free radical polymn. (NMFRP) was employed for various reasons: reduces presence of potentially cytotoxic organometallic catalysts common in atom transfer radical polymn. (ATRP); and it allows a phosphonate end-group instead of the common brominated end-group. Thermally oxidized silicon wafers were covalently functionalized with diethyl-(1-(N-(1-(3-(trimethoxysilyl)propylcarbamoyl)ethoxy)-N-tert-butylamino)ethyl)phosphonate. NMFRP was used to graft zwitterionic carboxybetaine methacrylamide monomers of varying inter-charge sepn. The resulting thin films were characterized using Attenuated Total Reflectance-Fourier Transform IR (ATR-FTIR) and X-ray photoelectron (XPS) spectroscopy, ellipsometry, water contact angle anal., and thermo gravimetric anal. (TGA). The effect of spacer group on the surface charge d. was detd. using zeta potential techniques. It is thought that this stratagem will facilitate the ability to tailor systematically both the interior and terminal polymer properties, providing a platform for further understanding how these conditions affect protein adsorption as well as cell-surface interactions. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011.
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      Giesbers, M.; Marcelis, A. T. M.; Zuilhof, H. Simulation of XPS C1s Spectra of Organic Monolayers by Quantum Chemical Methods. Langmuir 2013, 29, 4782– 4788,  DOI: 10.1021/la400445s
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      Simulation of XPS C1s Spectra of Organic Monolayers by Quantum Chemical Methods
      Giesbers, Marcel; Marcelis, Antonius T. M.; Zuilhof, Han
      Langmuir (2013), 29 (15), 4782-4788CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      Several simple methods are presented and evaluated to simulate the X-ray photoelectron spectra (XPS) of org. monolayers and polymeric layers by d. functional theory (DFT) and second-order Moller-Plesset theory (MP2) in combination with a series of basis sets. The simulated carbon (C1s) XPS spectra as obtained via B3LYP/6-311G(d,p) or M11/6-311G(d,p) calcns. are in good agreement (av. mean error <0.3 eV) with the exptl. spectra, and good ests. of C1s spectra can be obtained via EC1s(exp) = 0.9698EC1s(theory) + 20.34 (in eV) (B3LYP/6-311G(d,p)). As a result, the simulated C1s XPS spectra can elucidate the binding energies of the different carbon species within an org. layer and, in this way, greatly aid the assignment of complicated C1s XPS spectra. A wide range of examples, including haloalkanes, esters, (thio-)ethers, leaving groups, clickable functionalities, and bioactive moieties, are given.
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      Zhao, J.; Gao, F.; Pujari, S. P.; Zuilhof, H.; Teplyakov, A. V. Universal Calibration of Computationally Predicted N 1s Binding Energies for Interpretation of XPS Experimental Measurements. Langmuir 2017, 33, 10792– 10799,  DOI: 10.1021/acs.langmuir.7b02301
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      Universal Calibration of Computationally Predicted N 1s Binding Energies for Interpretation of XPS Experimental Measurements
      Zhao, Jing; Gao, Fei; Pujari, Sidharam P.; Zuilhof, Han; Teplyakov, Andrew V.
      Langmuir (2017), 33 (41), 10792-10799CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      Computationally predicted N 1s core level energies are commonly used to interpret the exptl. measurements obtained with XPS. This work compares the application of Koopmans' theorem to core electrons using the B3LYP functional with 2 commonly used basis sets, analyzes the factors relevant to the comparison of the computational with exptl. data, and presents several correlations that allow an accurate prediction of the N 1s binding energy. The 1st correlation is obtained with known N-contg. functional groups on well-characterized org. monolayers. This approach can then be reliably extended to a no. of N-contg. chem. systems on Si surfaces in which the nature of the chem. environment of N atoms had only been proposed based on a no. of anal. techniques. In most of those cases, the XPS anal. is consistent with the proposed structures, but is not always sufficient for conclusive assignments. Third, it was attempted to also include N-contg. systems on metals. Despite the admittedly oversimplified approach taken in this case (the metal surface is approximated by a single atom), the obsd. correlations are still exptl. useful, although in this case significant outliers are found. Finally, previously published correlations between exptl. and theor. C 1s data were reexamd., yielding a set of correlations that allow experimentalists to predict C 1s and N 1s XPS spectra with high accuracy.
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      Zhang, L.; Cao, Z.; Bai, T.; Carr, L.; Ella-Menye, J.-R.; Irvin, C.; Ratner, B. D.; Jiang, S. Zwitterionic hydrogels implanted in mice resist the foreign-body reaction. Nat. Biotechnol. 2013, 31, 553– 556,  DOI: 10.1038/nbt.2580
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      Zwitterionic hydrogels implanted in mice resist the foreign-body reaction
      Zhang, Lei; Cao, Zhiqiang; Bai, Tao; Carr, Louisa; Ella-Menye, Jean-Rene; Irvin, Colleen; Ratner, Buddy D.; Jiang, Shaoyi
      Nature Biotechnology (2013), 31 (6), 553-556CODEN: NABIF9; ISSN:1087-0156. (Nature Publishing Group)
      The performance of implantable biomedical devices is impeded by the foreign-body reaction, which results in formation of a dense collagenous capsule that blocks mass transport and/or elec. communication between the implant and the body. No known materials or coatings can completely prevent capsule formation. Here we demonstrate that ultra-low-fouling zwitterionic hydrogels can resist the formation of a capsule for at least 3 mo after s.c. implantation in mice. Zwitterionic hydrogels also promote angiogenesis in surrounding tissue, perhaps owing to the presence of macrophages exhibiting phenotypes assocd. with anti-inflammatory, pro-healing functions. Thus, zwitterionic hydrogels may be useful in a broad range of applications, including generation of biocompatible implantable medical devices and tissue scaffolds.
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      Tiwari, A.; Devi, P. P.; Yogesh, C.; Minakshi, P.; Sagar, M. G. Survival of Two Avian Respiratory Viruses on Porous and Nonporous Surfaces. Avian Dis. 2006, 50, 284– 287,  DOI: 10.1637/7453-101205R.1
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      Survival of two avian respiratory viruses on porous and nonporous surfaces
      Tiwari A; Patnayak Devi P; Chander Yogesh; Parsad Minakshi; Goyal Sagar M
      Avian diseases (2006), 50 (2), 284-7 ISSN:0005-2086.
      The transmission of pathogens from infected to susceptible hosts may occur through contaminated fomites and inanimate objects. This type of transmission depends on the ability of the pathogens to survive in the environment. In this report, we describe the survivability of two avian respiratory viruses, e.g., avian metapneumovirus and avian influenza virus on 12 different porous and nonporous surfaces. The viruses survived on some of the surfaces for up to 6 days postcontamination but not after 9 days. Both viruses survived longer on nonporous surfaces than on porous ones. One of the reasons for poor survival on porous surfaces could be inefficient elution of virus from these surfaces. These results should be helpful in determining how long the premises should be left vacant after an outbreak of these viruses has occurred in poultry houses.
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    • 1H NMR spectrum of N-[3-(decyldimethyl)-aminopropyl]methacrylamide bromide (A10); scale-up photoreactor scheme; simulated C 1s XPS spectra; wide-scan XPS spectra of the silicon and copper surfaces used in antiviral tests and AFM topography; viable viral particles of SARS-CoV-2 on different surfaces at different periods of time; viable viral particles of avian influenza on different surfaces at different periods of time (PDF)


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