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新型基因载体——修饰阳离子脂质体-聚乙烯亚胺-质粒纳米复合物的制备及其体外转染效率评估

Preparation and in-vitro transfection efficiency evaluation of modified cationic liposome-polyethyleneimine-plasmid nanocomplexes as a novel gene carrier.

作者信息

Mahmoudi Asma, Oskuee Reza Kazemi, Ramezani Mohammad, Malaekeh-Nikoue Bizhan

机构信息

School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, P.O. Box: 91775-1365, Iran.

出版信息

Curr Drug Deliv. 2014;11(5):636-42. doi: 10.2174/1567201811666140616160237.

Abstract

Liposome-linear polyethyleneimine (PEI)-DNA nanocomplexes have shown to be effective non-viral gene delivery vectors. In the present study, we tried to improve the transfection efficiency of these nanocomplexes by liposome modification. For this purpose, the lipopolymer was prepared by the conjugation of hexylacrylate to the PEI. Liposomes comprising lipopolymer and DOTAP (1.2-DiOleoyl-3-Trimethyl Ammonium-Propane) were prepared and extruded through polycarbonate filters to obtain the desired size. The 2.5, 25 and 250 KDa molecular weights of linear PEI have been used in order to prepare modified liposome-PEI-DNA nanocomplexes. Three C/P ratios of each nancomplex were premixed. Size, zeta potential and the DNA condensation ability of these complexes were determined separately, and in the end, the transfection efficiency and cell cytotoxicity of prepared vectors were evaluated on Neuro2A cell line. Mean particle size of all of these nanocomplexes was lower than 220 nm with surface charge of 17.5 to 25.9 mV. The lipopolyplexes (comprising modified liposome:PEI:DNA), modified liposome (as lipoplex) and PEI 250KDa (as polyplex) showed the highest transfection efficacy. This activity was amplified by increase carrier to plasmid (C/P) ratio. In addition, the metabolic activity of prepared vectors was 80-100% for control group. In conclusion, the prepared lipopolyplexes showed high ability to enhance gene transfer.

摘要

脂质体-线性聚乙烯亚胺(PEI)-DNA纳米复合物已被证明是有效的非病毒基因传递载体。在本研究中,我们试图通过脂质体修饰来提高这些纳米复合物的转染效率。为此,通过将丙烯酸己酯与PEI共轭制备了脂质聚合物。制备了包含脂质聚合物和DOTAP(1,2-二油酰基-3-三甲基铵丙烷)的脂质体,并通过聚碳酸酯滤膜挤出以获得所需尺寸。为了制备修饰的脂质体-PEI-DNA纳米复合物,使用了2.5、25和250 kDa分子量的线性PEI。将每种纳米复合物的三个C/P比预先混合。分别测定了这些复合物的大小、zeta电位和DNA凝聚能力,最后,在Neuro2A细胞系上评估了所制备载体的转染效率和细胞毒性。所有这些纳米复合物的平均粒径均低于220 nm,表面电荷为17.5至25.9 mV。脂质聚合物复合物(包含修饰的脂质体:PEI:DNA)、修饰的脂质体(作为脂质复合物)和PEI 250KDa(作为聚合物)显示出最高的转染效果。这种活性通过增加载体与质粒(C/P)比而增强。此外,所制备载体的代谢活性对于对照组为80-100%。总之,所制备的脂质聚合物复合物显示出高的增强基因转移能力。

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