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

Authors Yao JJ, Du YZ, Yuan H, You J, Hu FQ

Received 21 January 2014

Accepted for publication 19 February 2014

Published 18 June 2014 Volume 2014:9(1) Pages 2993—3003

DOI https://doi.org/10.2147/IJN.S61103

Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 3

Jing-Jing Yao, Yong-Zhong Du, Hong Yuan, Jian You, Fu-Qiang Hu

College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, People’s Republic of China

Abstract: 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.

Keywords: chitosan-g-stearic acid, cis-aconitate, micelles, transfection efficiency, intracellular trafficking

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