Nano Archive

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

Gao, Yu and Gu, Wangwen and Chen, Lingli and Xu, Zhenghong and Li, Yaping (2007) A multifunctional nano device as non-viral vector for gene delivery: In vitro characteristics and transfection. Journal of Controlled Release, 118 (3). 381 - 388.

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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(H2NPEGCA-co-HDCA)) to synthesize poly(Folate-HNPEGCA-co-HDCA), protamine 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 ( 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 (pH 4.5), but with 21.4% and 8.1% pDNA/PS release at PBS with pH 5.8 and 7.4 within 24 h, 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 1 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 MND 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™, 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.

Item Type:Article
Uncontrolled Keywords:Non-viral vector; pDNA; Multifunctional nano device; Folate; Protamine; Poly(cyanoacrylate)
Subjects:Biomedical Science > Nanomedicine
ID Code:5723
Deposited By:SPI
Deposited On:27 Jul 2009 10:11
Last Modified:27 Jul 2009 10:11

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