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聚(L-乳酸)/聚乙烯亚胺纳米颗粒作为质粒DNA载体

Poly(L-lactic acid)/polyethylenimine nanoparticles as plasmid DNA carriers.

作者信息

Park Yu-Mi, Shin Boo-Ahn, Oh In-Joon

机构信息

College of Pharmacy, Research Institute of Drug Development, Chonnam National University, Bug-gu, Gwangju 500-757, Korea.

出版信息

Arch Pharm Res. 2008 Jan;31(1):96-102. doi: 10.1007/s12272-008-1126-5.

Abstract

Non-viral vectors such as liposomes, polycations, and nanoparticles have been used as gene delivery systems. In this study, we prepared and characterized biodegradable poly(L-lactic acid) (PLA)/polyethylenimine (PEI) nanoparticles as gene carriers. pCMV/beta-gal and pEGFP-C1 were utilized as model plasmid DNAs (pDNA). Nanoparticles were prepared using a double emulsion-solvent evaporation technique, and their pDNA binding capacity was assessed by agarose gel electrophoresis. Transfection was studied in HEK 293 and HeLa cell lines, and the transfection efficiencies were determined by beta-galactosidase assay or flow cytometry. Three kinds of PLA/PEI systems were studied by varying the molecular weight of PEI. The PLA/PEI 25K system had a higher transfection efficiency than the PLA/PEI 0.8K or PLA/PEI 750K systems. The transfection efficiency was found to be dependent on the ratio of PLA/PEI nanoparticles to pDNA with an optimum ratio of 60:1 (w/w). The cytotoxicity was dependent on the quantity of PLA/PEI nanoparticles used, but it was comparable to that of commercial Lipofectin. These results demonstrate the potential of PLA/PEI nanoparticles as gene carriers.

摘要

脂质体、聚阳离子和纳米颗粒等非病毒载体已被用作基因递送系统。在本研究中,我们制备并表征了可生物降解的聚(L-乳酸)(PLA)/聚乙烯亚胺(PEI)纳米颗粒作为基因载体。pCMV/β-半乳糖苷酶和pEGFP-C1用作模型质粒DNA(pDNA)。采用复乳-溶剂蒸发技术制备纳米颗粒,并通过琼脂糖凝胶电泳评估其pDNA结合能力。在HEK 293和HeLa细胞系中研究转染情况,并通过β-半乳糖苷酶测定或流式细胞术确定转染效率。通过改变PEI的分子量研究了三种PLA/PEI体系。PLA/PEI 25K体系比PLA/PEI 0.8K或PLA/PEI 750K体系具有更高的转染效率。发现转染效率取决于PLA/PEI纳米颗粒与pDNA的比例,最佳比例为60:1(w/w)。细胞毒性取决于所使用的PLA/PEI纳米颗粒的数量,但与市售脂质体转染试剂相当。这些结果证明了PLA/PEI纳米颗粒作为基因载体的潜力。

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