College of Life Science and Technology, Huazhong University of Science and Technology, Luoyu Road 1037, Wuhan 430074, China.
J Biosci Bioeng. 2011 Jan;111(1):98-103. doi: 10.1016/j.jbiosc.2010.08.017. Epub 2010 Sep 19.
To enhance the transfection efficiency of the cationic liposomes, chitosan was selected as a DNA condensing agent. The particle size of the liposome/chitosan/DNA ternary complexes decreased and the zeta potential increased with the addition of chitosan. The formation of the ternary complexes was identified using agarose gel retardation study. The interaction of the ternary complexes was further confirmed by the decrease of the DNA fluorescence in the presence of Ru(phen)2dppz. In vitro and in vivo transfection activities of the complexes were determined using green fluorescent protein (GFP) expression in various cell lines and mouse tibial anterior muscle subcutaneously, respectively. Liposome/chitosan/DNA ternary complexes showed improved transfection efficiency in vitro cell culture system in the presence or absence of serum as well as in vivo mouse model system, as compared with liposome/DNA lipoplex. More importantly, the cell toxicity of the ternary complex is lower than that of lipoplex and liposome/poly-l-lysine/DNA ternary complex. The precondensation of DNA with chitosan can be a promising approach to further increase the transfection efficiency of cationic liposomes in clinical application.
为了提高阳离子脂质体的转染效率，选择壳聚糖作为 DNA 凝聚剂。随着壳聚糖的加入，脂质体/壳聚糖/DNA 三元复合物的粒径减小，zeta 电位增加。通过琼脂糖凝胶阻滞研究鉴定了三元复合物的形成。通过 Ru(phen)2dppz 的存在下 DNA 荧光的减少进一步证实了三元复合物的相互作用。通过在各种细胞系中 GFP 表达和小鼠胫骨前肌皮下转染复合物来确定复合物的体外和体内转染活性。与脂质体/DNA 脂质体相比，脂质体/壳聚糖/DNA 三元复合物在有或无血清的体外细胞培养系统以及体内小鼠模型系统中均显示出提高的转染效率。更重要的是，三元复合物的细胞毒性低于脂质体和脂质体/聚-l-赖氨酸/DNA 三元复合物。DNA 与壳聚糖的预凝聚可能是进一步提高阳离子脂质体在临床应用中转染效率的有前途的方法。
