Effects of incorporation of poly(γ-glutamic acid) in chitosan/DNA complex nanoparticles on cellular uptake and transfection efficiency

被引:105
作者
Peng, Shu-Fen [1 ]
Yang, Mei-Ju [1 ,2 ]
Su, Chun-Jen [1 ]
Chen, Hsin-Lung [1 ]
Lee, Po-Wei [1 ]
Wei, Ming-Cheng [1 ]
Sung, Hsing-Wen [1 ]
机构
[1] Natl Tsing Hua Univ, Bioengn Program, Dept Chem Engn, Hsinchu 30013, Taiwan
[2] Ind Technol Res Inst, Biomed Engn Res Labs, Hsinchu, Taiwan
关键词
Chitosan; Poly(gamma-glutamic acid); Transfection efficiency; Cellular uptake; MEDIATED GENE-TRANSFER; PLASMID DNA; DELIVERY; CARRIER; POLYSACCHARIDE; DEACETYLATION; PROTEOGLYCANS; ALGINATE; VECTORS;
D O I
10.1016/j.biomaterials.2008.12.019
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
Chitosan (CS)/DNA complex nanoparticles (NPs) have been considered as a vector for gene delivery. Although advantageous for DNA packing and protection, CS-based complexes may lead to difficulties in DNA release once arriving at the site of action. In this study, an approach through modifying their internal structure by incorporating a negatively charged poly(gamma-glutamic acid) (gamma-PGA) in CS/DNA complexes (CS/DNA/gamma-PGA NPs) is reported. The analysis of small angle X-ray scattering results revealed that DNA and gamma-PGA formed complexes with CS separately to yield two types of domains, leading to the formation of "compounded NPs". With this internal structure, the compounded NPs might disintegrate into a number of even smaller sub-particles after cellular internalization, thus improving the dissociation capacity of CS and DNA. Consequently, after incorporating gamma-PGA in CS/DNA complexes, a significant increase in their transfection efficiency was found. Interestingly, in addition to improving the release of DNA intracellularly, the incorporation of gamma-PGA in CS/DNA complexes significantly enhanced their cellular uptake. We further demonstrated that besides a non-specific charged-mediated binding to cell membranes, there were specific trypsin-cleavable proteins involved in the internalization of CS/DNA/gamma PGA NPs. The aforementioned results indicated that gamma-PGA played multiple important roles in enhancing the cellular uptake and transfection efficiency of CS/DNA/gamma-PGA NPs. (C) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1797 / 1808
页数:12
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