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Cationic Liposomes Modified with Polyallylamine as a Gene Carrier: Preparation, Characterization and Transfection Efficiency Evaluation

机译:以聚烯丙胺为基因载体修饰的阳离子脂质体的制备,表征和转染效率评价

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Purpose: Cationic polymers and cationic liposomes have shown to be effective non-viral gene delivery vectors. In this study, we tried to improve the transfection efficiency by employing the advantages of both. Methods: For this purpose, modified polyallylamines (PAAs) were synthesized. These modifications were done through the reaction of PAA (15 KDa) with acrylate and 6-bromoalkanoic acid derivatives. Liposomes comprising of these cationic polymers and cationic lipid were prepared and extruded through polycarbonate filters to obtain desired size. Liposome-DNA nanocomplexes were prepared in three carrier to plasmid (C/P) ratios. Size, zeta potential and DNA condensation ability of each complex were characterized separately and finally transfection efficiency and cytotoxicity of prepared vectors were evaluated in Neuro2A cell line. Results: The results showed that mean particle size of all these nanocomplexes was lower than 266 nm with surface charge of 22.0 to 33.9 mV. Almost the same condensation pattern was observed in all vectors and complete condensation was occurred at C/P ratio of 1.5. The lipoplexes containing modified PAA 15 kDa with 10% hexyl acrylate showed the highest transfection efficacy and lowest cytotoxicity in C/P ratio of 0.5. Conclusion: In some cases nanocomplexes consisting of cationic liposome and modified PAA showed better transfection activity and lower cytotoxicity compared to PAA.
机译:目的:阳离子聚合物和阳离子脂质体已被证明是有效的非病毒基因递送载体。在这项研究中,我们试图利用两者的优势来提高转染效率。方法:为此目的,合成了改性聚烯丙胺(PAA)。这些修饰是通过PAA(15 KDa)与丙烯酸酯和6-溴链烷酸衍生物的反应完成的。制备包含这些阳离子聚合物和阳离子脂质的脂质体,并通过聚碳酸酯过滤器挤出以获得所需的大小。以三种载体与质粒(C / P)的比例制备脂质体-DNA纳米复合物。分别表征每种复合物的大小,ζ电势和DNA缩合能力,最后在Neuro2A细胞系中评估制备的载体的转染效率和细胞毒性。结果:结果表明,所有这些纳米复合物的平均粒径均小于266 nm,表面电荷为22.0至33.9 mV。在所有载体中观察到几乎相同的缩合模式,并且在C / P比为1.5时发生了完全缩合。含修饰的PAA 15 kDa和10%丙烯酸己酯的脂质复合物在0.5的C / P比下显示出最高的转染效率和最低的细胞毒性。结论:在某些情况下,与阳离子脂质体相比,由阳离子脂质体和修饰的PAA组成的纳米复合物表现出更好的转染活性和较低的细胞毒性。

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