A multifunctional nano device as non-viral vector for gene delivery: In vitro characteristics and transfection

被引:27
作者
Gao, Yu [1 ]
Gu, Wangwen [1 ]
Chen, Lingli [1 ]
Xu, Zhenghong [1 ]
Li, Yaping [1 ]
机构
[1] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Mat Med, Shanghai 201203, Peoples R China
基金
中国国家自然科学基金;
关键词
non-viral vector; pDNA; multifunctional nano device; folate; protamine; poly(cyanoacrylate);
D O I
10.1016/j.jconrel.2007.01.006
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The aim of this work was to design a new multifunctional nano device (MND) for gene delivery. This MND was equipped with folic acid as ligand, which was conjugated to terminal amido of poly(aminopoly(ethylene glycol)cyanoacrylate-co-hexadecyl cyanoacrylate) (poly (H(2)NPEGCA-co-HDCA)) to synthesize poly(Folate-HNPEGCA-co-HDCA), prolamine sulfate (PS) as DNA condenser and for nuclear transfer, PEG chain from poly(Folate-HNPEGCA-co-HDCA) for decreasing macrophages recognition and extending half-life, dioleoyl phosphatidylethanolamine (DOPE) for endosomal escape, and we supposed that the latent DOPE fusogenicity could be gently restored along with fast degradation of poly(Folate-HNPEGCA-co-HDCA) in MND membrane within endosome. Our experimental results showed that optimum complexation (similar to 97%) of DNA was achieved at DNA:PS = 1:3 (w/w). The MND showed different loading ratio by lipid film hydration technique with the highest loading ratio about 12%, the particle size range 200-400 nm, surface charge range 8 mV-15 mV. MND1 (poly(Folate-HNPEGCA-co-HDCA)/DOPE, 5:95, molar ratio) exhibited a high burst release effect with 60% of pDNA/PS released within 1 day at PBS (pH4.5), but with 21.4% and 8.1% pDNA/PS release at PBS with pH 5.8 and 7.4 within 24 It, respectively. However, lesser pDNA/PS release occurred in MND2 (poly(Folate-HNPEGCA-co-HDCA)[DOPE, 10:90, molar ratio) with 46%, 16.9% and 7.8% of pDNA/PS released at PBS with pH 4.5, 5.8 and 7.4 within 24 h, respectively. After I day, pDNA/PS displayed a sustained release pattern. The amount of cumulated pDNA/PS release over 3 days was 75% and 51.2% at PBS with pH 4.5 for MND1 and MND2, respectively. The MD loading pDNA/PS showed that luciferase activity was over 0.5 ng luciferase/mg protein in KB cells, in particular, the MND1 showed the highest transfection efficiency (0.66 ng luciferase/mg protein) in KB cells, which was much higher compared with in A549 cells or other formulations such as LipofectAMINE (TM) free pDNA/PS and control multifunctional nano device (CMND), whose lipid film was consisted of poly(H2NPEGCA-co-HDCA) and DOPE. In addition, MND also showed good protection during encapsulation and low cytotoxicity. As a result, MND could be a more potential non-viral vector for delivery of DNA. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:381 / 388
页数:8
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