Layer-by-layer DNA films incorporating highly transfecting bioreducible poly(amido amine) and polyethylenimine for sequential gene delivery

• Published in: International journal of nanomedicine Vol. 13; pp. 4943 - 4960
• Main Authors: Xie, Lingxiao, Ding, Xiong, Budry, Rachel, Mao, Guangzhao
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 01.01.2018; Taylor & Francis Ltd; Dove Medical Press
• Subjects: Amines; Animals; Biocompatible Materials - chemistry; Biodegradable polymers; Cell Death; cell transfection; Copolymers; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA - chemistry; Fibroblasts; Genes; HEK293 Cells; Humans; Hydrodynamics; interlayer diffusion; localized gene delivery; Mice; Microscopy, Atomic Force; Molecular weight; NIH 3T3 Cells; Original Research; Particle Size; Polyamines - chemical synthesis; Polyamines - chemistry; polyelectrolyte multilayers; Polyelectrolytes; Polyethyleneimine - chemical synthesis; Polyethyleneimine - chemistry; Polymer industry; Polymerization; Proteins; sequential release; Solutions; Technology; Time-Lapse Imaging; Transfection; United States > US; cell transfection; polyelectrolyte multilayers; interlayer diffusion; sequential release; localized gene delivery; biodegradable polymers
• ISSN: 1178-2013; 1176-9114; 1178-2013
• DOI: 10.2147/ijn.s162353

The layer-by-layer (LbL) assembly method offers a molecular level control of the amount and spatial distribution of bioactive molecules. However, successful clinical translation of LbL film technology will most certainly require a better understanding and control of not only the film assembly process, but also film disassembly kinetics in physiologic conditions. This work focuses on the understanding and control of degradation properties of LbL films for localized gene delivery. Bioreducible poly(amido amine)s (PAAs) containing cystaminebisacrylamide (CBA), methylenebisacrylamide, and 5-amino-1-pentanol (APOL) were synthesized by Michael addition polymerization for the construction of bioreducible LbL films capable of sequential gene delivery. The synthesized PAAs were screened for desirable buffering capacity, cell transfection, and cytotoxicity characteristics together with 25 kDa branched polyethylenimine (PEI) and cross-linked 800 Da PEI. By screening the various polycations we were able to identify a copolymer of CBA and APOL for the subsequent construction of the LbL films. By incorporating a highly transfecting polycation and a nondiffusing polycation we were able to improve the overall transfection of HEK293 and MC3T3 cells from the bioreducible LbL films. We also demonstrated the dual-stage release and transfection of two different DNAs from the LbL films. The results indicate that LbL films consisting of bioreducible PAAs and non-diffusing polyelectrolytes have excellent degradation properties for the development of LbL coating technology for localized gene delivery applications.