ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Microfluidic Assembly of pDNA/Cationic Liposome Lipoplexes with High pDNA Loading for Gene Delivery

View Author Information
School of Chemical Engineering, University of Campinas, UNICAMP, Campinas, SP 13083-970, Brazil
Brazilian Synchrotron Light Laboratory, CNPEM, Campinas, São Paulo 13083-100, Brazil
§ Institute of Physics, University of São Paulo, USP, São Paulo, SP 05508, Brazil
Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas, Campinas, SP 13083-970, Brazil
*E-mail: latorre@feq Telephone: +55 19 35210397. Fax: +55 19 35213910.
Cite this: Langmuir 2016, 32, 7, 1799–1807
Publication Date (Web):January 27, 2016
Copyright © 2016 American Chemical Society

    Article Views





    Other access options


    Abstract Image

    Microfluidics offers unique characteristics to control the mixing of liquids under laminar flow. Its use for the assembly of lipoplexes represents an attractive alternative for the translation of gene delivery studies into clinical trials on a sufficient throughput scale. Here, it was shown that the microfluidic assembly of pDNA/cationic liposome (CL) lipoplexes allows the formation of nanocarriers with enhanced transfection efficiencies compared with the conventional bulk-mixing (BM) process under high pDNA loading conditions. Lipoplexes generated by microfluidic devices exhibit smaller and more homogeneous structures at a molar charge ratio (R±) of 1.5, representing the ratio of lipid to pDNA content. Using an optimized model to fit small-angle X-ray scattering (SAXS) curves, it was observed that large amounts of pDNA induces the formation of aggregates with a higher number of stacked bilayers (N ∼ 5) when the BM process was used, whereas microfluidic lipoplexes presented smaller structures with a lower number of stacked bilayers (N ∼ 2.5). In vitro studies further confirmed that microfluidic lipoplexes achieved higher in vitro transfection efficiencies in prostate cancer cells at R ± 1.5, employing a reduced amount of cationic lipid. The correlation of mesoscopic characteristics with in vitro performance provides insights for the elucidation of the colloidal arrangement and biological behavior of pDNA/CL lipoplexes obtained by different processes, highlighting the feasibility of applying microfluidics to gene delivery.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Cited By

    This article is cited by 35 publications.

    1. Sung Hwa Hong, Twinkal Patel, Shell Ip, Shyam Garg, Jung Kwon Oh. Microfluidic Assembly To Synthesize Dual Enzyme/Oxidation-Responsive Polyester-Based Nanoparticulates with Controlled Sizes for Drug Delivery. Langmuir 2018, 34 (10) , 3316-3325.
    2. Valeria Nele, Virginia Campani, Seyedeh Alia Moosavian, Giuseppe De Rosa. Lipid nanoparticles for RNA delivery: Self-assembling vs driven-assembling strategies. Advanced Drug Delivery Reviews 2024, 208 , 115291.
    3. Sima Mehraji, Don L. DeVoe. Microfluidic synthesis of lipid-based nanoparticles for drug delivery: recent advances and opportunities. Lab on a Chip 2024, 24 (5) , 1154-1174.
    4. Shirong Song, Zhikai Liu, Letao Guo, Wang Yao, Hongchen Liu, Mei Yang, Guangwen Chen. Continuous and size-control synthesis of lipopolyplex nanoparticles enabled by controlled micromixing performance for mRNA delivery. Journal of Flow Chemistry 2024, 15
    5. Saurabh Shah, Paras Famta, Dadi A. Srinivasarao, Syed Shahrukh, Naitik Jain, Akshay Shinde, Sajja Bhanu Prasad, Ganesh Vambhurkar, Giriraj Pandey, Rajendra Kumar, Shailendra Saraf, Saurabh Srivastava. Nanomedicine-RNAi interface: Architypes in prostate cancer therapeutics. Journal of Drug Delivery Science and Technology 2024, 92 , 105349.
    6. Rosilene Andrea Welter, Harrson Silva Santana, Lucimara Gaziola de la Torre, Mark C. Barnes, Osvaldir Pereira Taranto, Michael Oelgemöller. Biodiesel Production by Heterogeneous Catalysis and Eco‐friendly Routes. ChemBioEng Reviews 2023, 10 (2) , 86-111.
    7. Sofia Shtykalova, Dmitriy Deviatkin, Svetlana Freund, Anna Egorova, Anton Kiselev. Non-Viral Carriers for Nucleic Acids Delivery: Fundamentals and Current Applications. Life 2023, 13 (4) , 903.
    8. Nayab Tahir, Fatemeh Sharifi, Tanveer Ahmed Khan, Muhammad Muzammil Khan, Asadullah Madni, Mubashar Rehman. Microfluidics: A versatile tool for developing, optimizing, and delivering nanomedicines. 2023, 137-160.
    9. Yujie Yang, Zhen Liu, Hongchao Ma, Meiwen Cao. Application of Peptides in Construction of Nonviral Vectors for Gene Delivery. Nanomaterials 2022, 12 (22) , 4076.
    10. Harrson S. Santana, Victória A. Haddad, Paulo V. C. Calvo, Mauri S. A. Palma, Adriano G. P. da Silva, Dirceu Noriler, Osvaldir P. Taranto, João L. Silva. Design, optimization and scale-up of a new micromixer design based on plate column for organic synthesis. Chemical Engineering Journal 2022, 446 , 137159.
    11. Abdul Rahim Ferhan, Soohyun Park, Hyeonjin Park, Hyunhyuk Tae, Joshua A. Jackman, Nam‐Joon Cho. Lipid Nanoparticle Technologies for Nucleic Acid Delivery: A Nanoarchitectonics Perspective. Advanced Functional Materials 2022, 32 (37)
    12. Sara Gimondi, Carlos F. Guimarães, Sara F. Vieira, Virgínia M.F. Gonçalves, Maria E. Tiritan, Rui L. Reis, Helena Ferreira, Nuno M. Neves. Microfluidic mixing system for precise PLGA-PEG nanoparticles size control. Nanomedicine: Nanotechnology, Biology and Medicine 2022, 40 , 102482.
    13. Bruna G. Carvalho, Bruno T. Ceccato, Mariano Michelon, Sang W. Han, Lucimara G. de la Torre. Advanced Microfluidic Technologies for Lipid Nano-Microsystems from Synthesis to Biological Application. Pharmaceutics 2022, 14 (1) , 141.
    14. Shintaro Fumoto, Tsuyoshi Yamamoto, Kazuya Okami, Yuina Maemura, Chisato Terada, Asako Yamayoshi, Koyo Nishida. Understanding In Vivo Fate of Nucleic Acid and Gene Medicines for the Rational Design of Drugs. Pharmaceutics 2021, 13 (2) , 159.
    15. Masoumehossadat Hosseini, Navid Rabiee, Mojtaba Bagherzadeh. Targeted delivery of nucleic acids using microfluidic systems. 2021, 289-318.
    16. Sepideh Ahmadi, Navid Rabiee, Mojtaba Bagherzadeh, Mahdi Karimi. Microfluidic devices for gene delivery systems. 2021, 187-208.
    17. Yunchun Zhao, Haili Zheng, Xiaorong Wang, Xiaoling Zheng, Yongquan Zheng, Yue Chen, Weidong Fei, Jiahuan Zhu, Wenxi Wang, Caihong Zheng. Preparation and Biological Property Evaluation of Novel Cationic Lipid-Based Liposomes for Efficient Gene Delivery. AAPS PharmSciTech 2021, 22 (1)
    18. Signe Tandrup Schmidt, Dennis Christensen, Yvonne Perrie. Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b. Pharmaceutics 2020, 12 (12) , 1237.
    19. Ricardo Gaspar, Filipe Coelho, Bruno F. B. Silva. Lipid-Nucleic Acid Complexes: Physicochemical Aspects and Prospects for Cancer Treatment. Molecules 2020, 25 (21) , 5006.
    20. Pirthi Pal Singh, Veena Vithalapuram, Sunita Metre, Ravinder Kodipyaka. Lipoplex-based therapeutics for effective oligonucleotide delivery: a compendious review. Journal of Liposome Research 2020, 30 (4) , 313-335.
    21. C. Has, P. Sunthar. A comprehensive review on recent preparation techniques of liposomes. Journal of Liposome Research 2020, 30 (4) , 336-365.
    22. Qingming Ma, Jie Cao, Yang Gao, Shangcong Han, Yan Liang, Tingting Zhang, Xinyu Wang, Yong Sun. Microfluidic-mediated nano-drug delivery systems: from fundamentals to fabrication for advanced therapeutic applications. Nanoscale 2020, 12 (29) , 15512-15527.
    23. João Lameu da Silva Júnior, Victória Anselmo Haddad, Osvaldir Pereira Taranto, Harrson Silva Santana. Design and Analysis of New Micromixers Based on Distillation Column Trays. Chemical Engineering & Technology 2020, 43 (7) , 1249-1259.
    24. Ismail Eş, Leonardo Jose Montebugnoli, Maria Fernanda P. Filippi, Antonio A. Malfatti-Gasperini, Allan Radaic, Marcelo Bispo de Jesus, Lucimara Gaziola de la Torre. High-throughput conventional and stealth cationic liposome synthesis using a chaotic advection-based microfluidic device combined with a centrifugal vacuum concentrator. Chemical Engineering Journal 2020, 382 , 122821.
    25. Marco César Prado Soares, Matheus Kauê Gomes, Egont Alexandre Schenkel, Matheus dos Santos Rodrigues, Carlos Kenichi Suzuki, Lucimara Gaziola de la Torre, Eric Fujiwara. EVALUATION OF SILICA NANOPARTICLE COLLOIDAL STABILITY WITH A FIBER OPTIC QUASI-ELASTIC LIGHT SCATTERING SENSOR. Brazilian Journal of Chemical Engineering 2019, 36 (4) , 1519-1534.
    26. Chao Liu, Qiang Feng, Jiashu Sun. Lipid Nanovesicles by Microfluidics: Manipulation, Synthesis, and Drug Delivery. Advanced Materials 2019, 31 (45)
    27. Guangsheng Luo, Le Du, Yujun Wang, Kai Wang. Recent developments in microfluidic device-based preparation, functionalization, and manipulation of nano- and micro-materials. Particuology 2019, 45 , 1-19.
    28. Gabriel Perli, Amanda C.S.N. Pessoa, Tiago A. Balbino, Lucimara G. de la Torre. Ionic strength for tailoring the synthesis of monomodal stealth cationic liposomes in microfluidic devices. Colloids and Surfaces B: Biointerfaces 2019, 179 , 233-241.
    29. Harrson S. Santana, João L. Silva, Osvaldir P. Taranto. Development of microreactors applied on biodiesel synthesis: From experimental investigation to numerical approaches. Journal of Industrial and Engineering Chemistry 2019, 69 , 1-12.
    30. Micaela Tamara Vitor, Sébastien Sart, Antoine Barizien, Lucimara Gaziola De La Torre, Charles N. Baroud. Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform. Scientific Reports 2018, 8 (1)
    31. Hao Zhang, Yifeng Zhu, Youqing Shen. Microfluidics for Cancer Nanomedicine: From Fabrication to Evaluation. Small 2018, 14 (28)
    32. Patrícia Severino, Elisânia F. Silveira, Kahynna Loureiro, Marco V. Chaud, Danilo Antonini, Marcelo Lancellotti, Victor Hugo Sarmento, Classius F. da Silva, Maria Helena A. Santana, Eliana B. Souto. Antimicrobial activity of polymyxin-loaded solid lipid nanoparticles (PLX-SLN): Characterization of physicochemical properties and in vitro efficacy. European Journal of Pharmaceutical Sciences 2017, 106 , 177-184.
    33. Tiago A. Balbino, Juliana M. Serafin, Allan Radaic, Marcelo B. de Jesus, Lucimara G. de la Torre. Integrated microfluidic devices for the synthesis of nanoscale liposomes and lipoplexes. Colloids and Surfaces B: Biointerfaces 2017, 152 , 406-413.
    34. Rafael F. Alves, Marianna T.P. Favaro, Tiago A. Balbino, Marcelo A.S. de Toledo, Lucimara G. de la Torre, Adriano R. Azzoni. Recombinant protein-based nanocarriers and their association with cationic liposomes: Characterization and in vitro evaluation. Colloids and Surfaces A: Physicochemical and Engineering Aspects 2017, 513 , 1-10.
    35. Chanuk Jeong, Jisang Yoo, DaeYong Lee, Yeu-Chun Kim. A branched TAT cell-penetrating peptide as a novel delivery carrier for the efficient gene transfection. Biomaterials Research 2016, 20 (1)

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect