Efficient gene delivery system mediated by cis-aconitate-modified chitosan-g-stearic acid micelles

Cis-aconitate-modified chitosan-g-stearic acid (CA-CSO-SA) micelles were synthesized in this study to improve the gene transfection efficiency of chitosan-g-stearic acid (CSO-SA). The CA-CSO-SA micelles had a similar size, critical micelle concentration, and morphology, but their zeta potential and cytotoxicity were reduced compared with CSO-SA micelles. After modification with cis-aconitate, the CA-CSO-SA micelles could also compact plasmid DNA (pDNA) to form nanocomplexes. However, the DNA binding ability of CA-CSO-SA was slightly reduced compared with that of CSO-SA. The transfection efficiency mediated by CA-CSO-SA/pDNA against HEK-293 cells reached up to 37%, and was much higher than that of CSO-SA/pDNA (16%). Although the cis-aconitate modification reduced cellular uptake kinetics in the initial stages, the total amount of cellular uptake tended to be the same after 24 hours of incubation. An endocytosis inhibition experiment showed that the internalization mechanism of CA-CSO-SA/pDNA in HEK-293 cells was mainly via clathrin-mediated endocytosis, as well as caveolae-mediated endocytosis and macropinocytosis. Observation of intracellular trafficking indicated that the CSO-SA/pDNA complexes were trapped in endolysosomes, but CA-CSO-SA/pDNA was more widely distributed in the cytosol. This study suggests that modification with cis-aconitate improves the transfection efficiency of CSO-SA/pDNA.

[1]  D. Peer,et al.  Polysaccharides as building blocks for nanotherapeutics. , 2012, Chemical Society reviews.

[2]  P. Lu,et al.  Quaternary complexes composed of plasmid DNA/protamine/fish sperm DNA/stearic acid grafted chitosan oligosaccharide micelles for gene delivery. , 2011, International journal of biological macromolecules.

[3]  Neil B. McKeown,et al.  Engineering of Dendrimer Surfaces to Enhance Transepithelial Transport and Reduce Cytotoxicity , 2003, Pharmaceutical Research.

[4]  L. Smith,et al.  Peptide-based gene delivery. , 1999, Current opinion in molecular therapeutics.

[5]  A S Hoffman,et al.  Molecular engineering of proteins and polymers for targeting and intracellular delivery of therapeutics. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[6]  Hong Yuan,et al.  Effect of proteins with different isoelectric points on the gene transfection efficiency mediated by stearic acid grafted chitosan oligosaccharide micelles. , 2013, Molecular pharmaceutics.

[7]  Changren Zhou,et al.  Polysaccharides-based nanoparticles as drug delivery systems. , 2008, Advanced drug delivery reviews.

[8]  M. Tabrizian,et al.  Cell line-dependent internalization pathways and intracellular trafficking determine transfection efficiency of nanoparticle vectors. , 2008, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[9]  T. Xia,et al.  Understanding biophysicochemical interactions at the nano-bio interface. , 2009, Nature materials.

[10]  Hong Yuan,et al.  Polymeric micelles with glycolipid-like structure and multiple hydrophobic domains for mediating molecular target delivery of paclitaxel. , 2007, Biomacromolecules.

[11]  M. Alonso,et al.  Hyaluronic Acid/Chitosan-g-Poly(ethylene glycol) Nanoparticles for Gene Therapy: An Application for pDNA and siRNA Delivery , 2010, Pharmaceutical Research.

[12]  P. Artursson,et al.  Molecular design of chitosan gene delivery systems with an optimized balance between polyplex stability and polyplex unpacking. , 2010, Biomaterials.

[13]  Hongmei Wu,et al.  Serum tolerance and endosomal escape capacity of histidine-modified pDNA-loaded complexes based on polyamidoamine dendrimer derivatives. , 2012, Biomaterials.

[14]  A. Hoffman,et al.  pH-sensitive polymers that enhance intracellular drug delivery in vivo. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[15]  Cui Tang,et al.  The effect of crosslinking agents on the transfection efficiency, cellular and intracellular processing of DNA/polymer nanocomplexes. , 2013, Biomaterials.

[16]  Bingqing Wang,et al.  Effects of hydrophobic and hydrophilic modifications on gene delivery of amphiphilic chitosan based nanocarriers. , 2011, Biomaterials.

[17]  K. Hamada,et al.  DNA/polyethyleneimine/hyaluronic acid small complex particles and tumor suppression in mice. , 2010, Biomaterials.

[18]  Shuming Nie,et al.  Proton-sponge coated quantum dots for siRNA delivery and intracellular imaging. , 2008, Journal of the American Chemical Society.

[19]  Wei Sun,et al.  Chitosan-based formulations for delivery of DNA and siRNA. , 2010, Advanced drug delivery reviews.

[20]  S. Kannan,et al.  The effect of surface functionality on cellular trafficking of dendrimers. , 2008, Biomaterials.

[21]  Hong Yuan,et al.  Cellular uptake and cytotoxicity of shell crosslinked stearic acid-grafted chitosan oligosaccharide micelles encapsulating doxorubicin. , 2008, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[22]  M. Buschmann,et al.  Intracellular trafficking and decondensation kinetics of chitosan-pDNA polyplexes. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[23]  F. Hu,et al.  Effective antitumor gene therapy delivered by polyethylenimine-conjugated stearic acid-g-chitosan oligosaccharide micelles , 2012, Gene Therapy.

[24]  P. Orlandi,et al.  Filipin-dependent Inhibition of Cholera Toxin: Evidence for Toxin Internalization and Activation through Caveolae-like Domains , 1998, The Journal of cell biology.

[25]  A. George,et al.  Gene delivery by dendrimers operates via a cholesterol dependent pathway. , 2004, Nucleic acids research.

[26]  J. Thomas,et al.  Polymer-induced leakage of cations from dioleoyl phosphatidylcholine and phosphatidylglycerol liposomes. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[27]  Gert Storm,et al.  Endosomal escape pathways for delivery of biologicals. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[28]  Jing-yu Zhang,et al.  Phosphorylatable short peptide conjugation for facilitating transfection efficacy of CS/DNA complex. , 2010, International journal of pharmaceutics.

[29]  N. Selvamurugan,et al.  Chitosan and its derivatives for gene delivery. , 2011, International journal of biological macromolecules.

[30]  Hsin‐Lung Chen,et al.  Enhancement of efficiencies of the cellular uptake and gene silencing of chitosan/siRNA complexes via the inclusion of a negatively charged poly(γ-glutamic acid). , 2010, Biomaterials.

[31]  Chih-Kuang Chen,et al.  Overcoming nonviral gene delivery barriers: perspective and future. , 2013, Molecular pharmaceutics.

[32]  I. Khalil,et al.  Uptake Pathways and Subsequent Intracellular Trafficking in Nonviral Gene Delivery , 2006, Pharmacological Reviews.

[33]  Hong Yuan,et al.  Synthesis and antitumor activity of doxorubicin conjugated stearic acid-g-chitosan oligosaccharide polymeric micelles. , 2009, Biomaterials.

[34]  Marie C. M. Lin,et al.  Nucleic acid delivery with chitosan and its derivatives. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[35]  Influence of the polyanion on the physico-chemical properties and biological activities of polyanion/DNA/polycation ternary polyplexes. , 2012, Acta biomaterialia.

[36]  Ü. Langel,et al.  Delivery of short interfering RNA using endosomolytic cell‐penetrating peptides , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[37]  Tomoko Ito,et al.  Novel poly(ethylene glycol) derivatives with carboxylic acid pendant groups: synthesis and their protection and enhancing effect on non-viral gene transfection systems , 2003, Journal of biomaterials science. Polymer edition.

[38]  Stephanie E. A. Gratton,et al.  The effect of particle design on cellular internalization pathways , 2008, Proceedings of the National Academy of Sciences.

[39]  Nobuhiro Nishiyama,et al.  Rational design of smart supramolecular assemblies for gene delivery: chemical challenges in the creation of artificial viruses. , 2012, Chemical Society reviews.

[40]  Hong Yuan,et al.  High cytotoxicity and resistant-cell reversal of novel paclitaxel loaded micelles by enhancing the molecular-target delivery of the drug , 2007, Nanotechnology.

[41]  H. Sung,et al.  The characteristics, cellular uptake and intracellular trafficking of nanoparticles made of hydrophobically-modified chitosan. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[42]  S. Zeng,et al.  A novel chitosan oligosaccharide-stearic acid micelles for gene delivery: properties and in vitro transfection studies. , 2006, International journal of pharmaceutics.