College of Pharmaceutical Science, Zhejiang University, 388 Yuhangtang Road, Hangzhou 310058, PR China.
Int J Biol Macromol. 2011 Jan 1;48(1):153-9. doi: 10.1016/j.ijbiomac.2010.10.011. Epub 2010 Oct 28.
Quaternary complexes with condensed core of plasmid DNA, protamine, fish sperm DNA and shell of stearic acid grafted chitosan oligosaccharide (CSO-SA), were prepared. The CSO-SA could self-assemble to form nano-sized micelles in aqueous solution and demonstrated excellent internalization ability of tumor cells. Dynamic light scattering (DLS) measurement and transmission electrostatic microscope (TEM) images showed that quaternary complexes had spherical shape with about 25 nm number average diameter, and the size of quaternary complexes was smaller than that of CSO-SA micelles and CSO-SA micelles/plasmid DNA binary complexes. The transfection efficiencies of quaternary complexes on HEK293 and MCF-7 cells increased with incubation time, and were significantly higher than that of CSO-SA micelles/plasmid DNA binary complexes. The optimal transfection efficiency of quaternary complexes on HEK293 and MCF-7 cells measured by flow cytometer after 96 h was 23.82% and 41.43%, respectively. Whereas, the transfection efficiency of Lipofectamine™ 2000 on HEK293 and MCF-7 cells after 96 h was 32.45% and 33.23%, respectively. The data of luciferease activity measurement showed that the optimal ratio of plasmid DNA:fish sperm DNA:protamine:CSO-SA was 1:1:5:5. The results indicated that the present quaternary complexes were potential non-viral gene delivery system.
制备了具有质粒 DNA、鱼精蛋白、鱼精 DNA 凝聚核心和接枝硬脂酸壳聚糖寡糖（CSO-SA）的季铵复合物。CSO-SA 可以自组装在水溶液中形成纳米级胶束，并显示出对肿瘤细胞的优异内化能力。动态光散射（DLS）测量和透射电子显微镜（TEM）图像表明，季铵复合物具有约 25nm 的数均直径的球形形状，并且季铵复合物的尺寸小于 CSO-SA 胶束和 CSO-SA 胶束/质粒 DNA 二元复合物的尺寸。季铵复合物在 HEK293 和 MCF-7 细胞上的转染效率随孵育时间的增加而增加，并且明显高于 CSO-SA 胶束/质粒 DNA 二元复合物。通过流式细胞仪测量，96 小时后季铵复合物对 HEK293 和 MCF-7 细胞的最佳转染效率分别为 23.82%和 41.43%。然而，Lipofectamine™2000 在 HEK293 和 MCF-7 细胞中的转染效率在 96 小时后分别为 32.45%和 33.23%。荧光素酶活性测量的数据表明，质粒 DNA:鱼精 DNA:鱼精蛋白:CSO-SA 的最佳比例为 1:1:5:5。结果表明，本季铵复合物是潜在的非病毒基因传递系统。
