School of Chemical Engineering and Technology, Tianjin University , Yaguan Road 135, Tianjin 300350, China.
School of Chemistry and Chemical Engineering, Qinghai University for Nationalities , Bayi middle Road 3, Xining, Qinghai 810007, China.
Langmuir. 2017 Nov 21;33(46):13315-13325. doi: 10.1021/acs.langmuir.7b02934. Epub 2017 Nov 8.
In order to improve the transfection efficiency and reduce the cytotoxicity of gene carriers, many strategies have been used to develop novel gene carriers. In this study, five complex micelles (MSP(2 k), MSP(4 k), MSP(6 k), MSP(8 k), and MSP(10 k)) were prepared from methoxy-poly(ethylene glycol)-b-poly(d,l-lactide-co-glycolide) (mPEG-b-PLGA) and sorbitol-poly(d,l-lactide-co-glycolide)-graft-PEI (sorbitol-PLGA-g-PEI, where the designed molecular weights of PLGA chains were 2 kDa, 4 kDa, 6 kDa, 8 kDa, and 10 kDa, respectively) copolymers by a self-assembly method, and the mass ratio of mPEG-b-PLGA to sorbitol-PLGA-g-PEI was 1/3. These complex micelles and their gene complexes had appropriate sizes and zeta potentials, and pEGFP-ZNF580 (pDNA) could be efficiently internalized into EA.hy926 cells by their gene complexes (MSP(2 k)/pDNA, MSP(4 k)/pDNA, MSP(6 k)/pDNA, MSP(8 k)/pDNA, and MSP(10 k)/pDNA). The MTT assay results demonstrated that the gene complexes had low cytotoxicity in vitro. When the hydrophobic PLGA chain increased above 6 kDa, the gene complexes showed higher performance than that prepared from short hydrophobic chains. Moreover, the relative ZNF580 protein expression levels in MSP(6 k)/pDNA, MSP(8 k)/pDNA, and MSP(10 k)/pDNA) groups were 79.6%, 71.2%, and 73%, respectively. These gene complexes could promote the transfection of endothelial cells, while providing important information and insight for the design of new and effective gene carriers to promote the proliferation and migration of endothelial cells.
为了提高基因载体的转染效率和降低细胞毒性，已经采用了许多策略来开发新型基因载体。在这项研究中，我们由甲氧基聚乙二醇-b-聚（d,l-丙交酯-co-乙交酯）（mPEG-b-PLGA）和山梨糖醇聚（d,l-丙交酯-co-乙交酯）-接枝-聚乙烯亚胺（sorbitol-PLGA-g-PEI，其中设计的 PLGA 链分子量分别为 2 kDa、4 kDa、6 kDa、8 kDa 和 10 kDa）共聚物通过自组装方法制备了 5 种复合胶束（MSP(2 k)、MSP(4 k)、MSP(6 k)、MSP(8 k)和 MSP(10 k)），mPEG-b-PLGA 与 sorbitol-PLGA-g-PEI 的质量比为 1/3。这些复合胶束及其基因复合物具有适当的粒径和 Zeta 电位，其基因复合物（MSP(2 k)/pDNA、MSP(4 k)/pDNA、MSP(6 k)/pDNA、MSP(8 k)/pDNA 和 MSP(10 k)/pDNA）可以有效地将 pEGFP-ZNF580（pDNA）转染进入 EA.hy926 细胞。MTT 检测结果表明，这些基因复合物在体外具有较低的细胞毒性。当疏水性 PLGA 链增加到 6 kDa 以上时，基因复合物的性能优于由短疏水性链制备的基因复合物。此外，MSP(6 k)/pDNA、MSP(8 k)/pDNA 和 MSP(10 k)/pDNA 组的相对 ZNF580 蛋白表达水平分别为 79.6%、71.2%和 73%。这些基因复合物可以促进内皮细胞的转染，同时为设计新的有效的基因载体以促进内皮细胞的增殖和迁移提供了重要的信息和见解。
