Sun Yun-Xia, Zhang Xian-Zheng, Cheng Han, Cheng Si-Xue, Zhuo Ren-Xi
Department of Chemistry, Key Laboratory of Biomedical Polymers of Ministry of Education, Wuhan University, Wuhan 430072, People's Republic of China.
J Biomed Mater Res A. 2008 Mar 15;84(4):1102-10. doi: 10.1002/jbm.a.31520.
In this study, the low molecular weight branched polyethylenimine (PEI) (800 Da PEI) was grafted to the biodegradable and biocompatible carboxymethyl dextran (CMD) to obtain CMD-g-PEI, and the plasmid DNA was complexed with CMD-g-PEI polycation to form the polyion complex. The acid-base titration profile showed that the CMD-g-PEI had endosomal disruption capacity, and the agarose gel electrophoresis suggested that the CMD-g-PEI could condense DNA efficiently. The transfection efficiency of CMD-g-PEI/DNA complexes was measured by luciferase and green fluorescent protein assay in HEK293 cells. The results revealed that the transfection efficiency of CMD-g-PEI at the N/P ratios over 30-70 was higher than or comparable to that of the 25 kDa PEI at optimal ratio (N/P = 10). The cytotoxicity of CMD-g-PEI as well as 25 kDa PEI was evaluated in NIH3T3 and HEK293 cells, respectively, and it was also found that the cytotoxicity of CMD-g-PEI was much lower than that of 25 kDa PEI. The resulted CMD-g-PEI with high transfection efficiency and low cytotoxicity have promising applications when used as gene vector.
在本研究中,将低分子量支链聚乙烯亚胺(PEI)(800 Da PEI)接枝到可生物降解且具有生物相容性的羧甲基葡聚糖(CMD)上,以获得CMD-g-PEI,然后将质粒DNA与CMD-g-PEI聚阳离子复合形成聚离子复合物。酸碱滴定曲线表明CMD-g-PEI具有内体破坏能力,琼脂糖凝胶电泳表明CMD-g-PEI能够有效地凝聚DNA。通过荧光素酶和绿色荧光蛋白测定法在HEK293细胞中测量CMD-g-PEI/DNA复合物的转染效率。结果显示,当N/P比超过30-70时,CMD-g-PEI的转染效率高于或与25 kDa PEI在最佳比例(N/P = 10)时相当。分别在NIH3T3和HEK293细胞中评估了CMD-g-PEI以及25 kDa PEI的细胞毒性,还发现CMD-g-PEI的细胞毒性远低于25 kDa PEI。所得到的具有高转染效率和低细胞毒性的CMD-g-PEI作为基因载体具有广阔的应用前景。