Arginine-chitosan/DNA self-assemble nanoparticles for gene delivery:: In vitro characteristics and transfection efficiency
Arginine-chitosan/DNA self-assemble nanoparticles for gene delivery:: In vitro characteristics and transfection efficiency
复制标题
用于基因传递的精氨酸-壳聚糖/DNA 自组装纳米颗粒：体外特性和转染效率。
DOI：
10.1016/j.ijpharm.2008.03.037
复制
发表时间：
2008-07-09
影响因子：
5.8
通讯作者：
Li, Yaping
中科院分区：
文献类型：
--
作者：
Gao, Yu;Xu, Zhenghong;Li, Yaping
壳聚糖(Cs)是一种天然阳离子多糖，由于其生物相容性好、毒性低，已显示出作为非病毒基因载体的潜力。然而，壳聚糖的水溶性差、转染率低，限制了其用于基因传递的研究。本工作的目的是制备精氨酸-壳聚糖(Arg-Cs)/DNA自组装纳米粒(ACSNs)，并测定其体外特性和对HEK 293和COS-7细胞的转染率。实验结果表明，不同N/P比制备的ACSNs的粒径为200~400 nm，Zeta电位为0.23~12.25 mV。ACSNs的体外转染率与转染液的pH值呈依赖关系，在pH值为7.2时表达效率最高。随着壳聚糖胺/DNA磷酸酯(N/P)的比例从1增加到5，转染率逐渐增加，当N/P比为5时，转染率达到最高水平。对HEK 293细胞和COS-7细胞，质粒剂量对转染率的影响显示，4µg/孔和6wg/孔的转染率最高。ACSNs的转染率远高于Cs/DNA自组装纳米粒(CSNS)。ACSNs的平均细胞存活率在90%以上。这些结果表明，ACSNs可能是一种安全有效的非病毒载体。(C)2008爱思唯尔B.V.保留所有权利。
Chitosan (Cs) is a natural cationic polysaccharide that has shown potential as non-viral vector for gene delivery because of its biocompatibility and low toxicity. However, chitosan used for gene delivery is limited due to its poorwater solubility and low transfection efficiency. The purpose of this work was to prepare Arginine-chitosan (Arg-Cs)/DNA self-assemble nanoparticles (ACSNs), and determine their in vitro characteristics and transfection efficiency against HEK 293 and COS-7 cells. Our experimental results showed that the particle size and zeta potential of ACSNs prepared with different N/P ratios were 200-400 nm and 0.23-12.25 mV, respectively. The in vitro transfection efficiency of ACSNs showed dependence on pH of transfection medium, and the highest expression efficiency was obtained at pH 7.2. The transfection efficiency increased with the ratio of chitosan-amine/DNA phosphate (N/P ratio) from 1 to 5, and reached the highest level with the N/P ratio 5. Effect of plasmid dosage on the transfection efficiency showed the highest transfection efficiency was obtained at 4 mu g/well for HEK 293 cells and 6 wg/well for COS-7 cells. The transfection efficiency of ACSNs was much higher than that of Cs/DNA self-assemble nanoparticles (CSNs). The average cell viability of ACSNs was over 90%. These results suggested that ACSNs could be a safe and effective non-viral vector for gene delivery. (C) 2008 Elsevier B.V. All rights reserved.