Charge shielding effects on gene delivery of polyethylenimine/DNA complexes: PEGylation and phospholipid coating

Polyethylenimine (PEI) is an efficient cationic polymer for gene delivery, but defective in biocompatibility. In this study, we developed two different strategies to shield the positively charged PEI/DNA complexes: PEGylation and lipid coating. The physicochemical properties, cytotoxicity and transfection efficiency of the two gene delivery systems were investigated. Both PEGylation and lipid coating succeeded in reducing the zeta-potential of the complexes. Lipid-coated PEI/DNA complexes (LPD complexes) and PEI/DNA complexes exhibited similar cytotoxicity, whereas PEG–PEI/DNA complexes showed lower cytotoxicity, especially at high N/P ratios. LPD complexes were less efficient in transfection compared to PEG–PEI/DNA complexes. The transfection efficiency was influenced remarkably by cytotoxicity and surface charge of the complexes. Intracellular processes studies revealed that endosomal release might be one of the rate-limiting steps in cell transfection with PEI as a gene delivery carrier.

[1]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Wei Zhang,et al.  Stability of poly(ethylene glycol)-graft-polyethylenimine copolymer/DNA complexes: influences of PEG molecular weight and PEGylation degree , 2010, Journal of materials science. Materials in medicine.

[3]  A. Mikos,et al.  Poly(ethylenimine) and its role in gene delivery. , 1999, Journal of controlled release : official journal of the Controlled Release Society.

[4]  J. Nah,et al.  Degradable polyethylenimine-alt-poly(ethylene glycol) copolymers as novel gene carriers. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[5]  D. Fischer,et al.  Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives , 2005, The journal of gene medicine.

[6]  S Moein Moghimi,et al.  A two-stage poly(ethylenimine)-mediated cytotoxicity: implications for gene transfer/therapy. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[7]  B. Sproat,et al.  Nonviral siRNA delivery to the lung: investigation of PEG-PEI polyplexes and their in vivo performance. , 2009, Molecular pharmaceutics.

[8]  R. Weissleder,et al.  Linear polyethyleneimine grafted to a hyperbranched poly(ethylene glycol)-like core: a copolymer for gene delivery. , 2006, Bioconjugate chemistry.

[9]  Y. Mély,et al.  Role of endocytosis in the transfection of L929 fibroblasts by polyethylenimine/DNA complexes. , 2001, Biochimica et biophysica acta.

[10]  Wim E Hennink,et al.  Lipid-coated polyplexes for targeted gene delivery to ovarian carcinoma cells , 2001, Cancer Gene Therapy.

[11]  Wenjin Guo,et al.  Efficient Gene Delivery Using Anionic Liposome-Complexed Polyplexes (LPDII) , 2000, Bioscience reports.

[12]  R. Müller,et al.  Transfection with different colloidal systems: comparison of solid lipid nanoparticles and liposomes. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[13]  Kyung-Dall Lee,et al.  Enhanced plasmid DNA delivery using anionic LPDII by listeriolysin O incorporation , 2005, The journal of gene medicine.

[14]  T. Chow,et al.  Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo , 2004, Gene Therapy.

[15]  Düzgüneş,et al.  Mechanisms and kinetics of liposome-cell interactions. , 1999, Advanced drug delivery reviews.

[16]  S. Carotta,et al.  Different behavior of branched and linear polyethylenimine for gene delivery in vitro and in vivo , 2001, The journal of gene medicine.

[17]  J. Heyes,et al.  Lipid encapsulation enables the effective systemic delivery of polyplex plasmid DNA. , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  W. Tseng,et al.  Improved stability of polycationic vector by dextran-grafted branched polyethylenimine. , 2003, Biomacromolecules.

[19]  W. Tseng,et al.  Dependence of transgene expression and the relative buffering capacity of dextran-grafted polyethylenimine. , 2005, Molecular pharmaceutics.

[20]  Y. T. Ko,et al.  Liposome encapsulated polyethylenimine/ODN polyplexes for brain targeting. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[21]  J. Behr,et al.  Effective polyethylenimine‐mediated gene transfer into human endothelial cells , 2004, The journal of gene medicine.

[22]  Clive J Roberts,et al.  Polyethylenimine-graft-poly(ethylene glycol) copolymers: influence of copolymer block structure on DNA complexation and biological activities as gene delivery system. , 2002, Bioconjugate chemistry.

[23]  Thomas Kissel,et al.  In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. , 2003, Biomaterials.

[24]  W. Hennink,et al.  Effects of Physicochemical Characteristics of Poly(2-(dimethylamino)ethyl methacrylate)-Based Polyplexes on Cellular Association and Internalization , 2000, Journal of drug targeting.

[25]  Xingen Luo,et al.  Poly(ethylene glycol)-block-polyethylenimine copolymers as carriers for gene delivery: effects of PEG molecular weight and PEGylation degree. , 2008, Journal of biomedical materials research. Part A.

[26]  A. Göpferich,et al.  Polyethylenimine-based non-viral gene delivery systems. , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[27]  G. Lutz,et al.  PEG-PEI copolymers for oligonucleotide delivery to cells and tissues. , 2008, Methods in molecular biology.

[28]  A. Kichler.  Gene transfer with modified polyethylenimines , 2004, The journal of gene medicine.

[29]  Jung-Ki Park,et al.  Effect of polyethylene glycol on gene delivery of polyethylenimine. , 2003, Biological & pharmaceutical bulletin.

[30]  D. Fischer,et al.  Low-molecular-weight polyethylenimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine. , 2003, Journal of controlled release : official journal of the Controlled Release Society.

[31]  M. Glodde,et al.  Physiochemical properties of low and high molecular weight poly(ethylene glycol)-grafted poly(ethylene imine) copolymers and their complexes with oligonucleotides. , 2006, Biomacromolecules.

[32]  A. Mikos,et al.  Size matters: molecular weight affects the efficiency of poly(ethylenimine) as a gene delivery vehicle. , 1999, Journal of biomedical materials research.

[33]  Wei Zhang,et al.  PEG- and PDMAEG-Graft-Modified Branched PEI as Novel Gene Vector: Synthesis, Characterization and Gene Transfection , 2010, Journal of biomaterials science. Polymer edition.

[34]  T. Bieber,et al.  Intracellular route and transcriptional competence of polyethylenimine-DNA complexes. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[35]  A. Kabanov,et al.  Design and formulation of polyplexes based on pluronic-polyethyleneimine conjugates for gene transfer. , 2002, Bioconjugate chemistry.