Department of Cardiac Surgery, Zhongshan Hospital, Fudan University & Shanghai Institute of Cardiovascular Diseases, Shanghai 200032, People's Republic of China.
J Mater Sci Mater Med. 2011 Nov;22(11):2477-85. doi: 10.1007/s10856-011-4424-2. Epub 2011 Aug 26.
We investigated the feasibility and efficacy of hyperbranched polyamidoamine (hPAMAM) mediated human vascular endothelial growth factor-165 (hVEGF(165)) gene transfer into skeletal myoblasts for cardiac repair. The hPAMAM was synthesized using a modified one-pot method. Encapsulated DNA was protected by hPAMAM from degradation for over 120 min. The transfection efficiency of hPAMAM in myoblasts was 82.6 ± 7.0% with cell viability of 94.6 ± 1.4% under optimal conditions. The hPAMAM showed much higher transfection efficiency (P < 0.05) than polyetherimide and Lipofectamine 2000 with low cytotoxicity. The transfected skeletal myoblasts gave stable hVEGF(165) expression for 18 days. After transplantation of hPAMAM-hVEGF(165) transfected cells, apoptotic myocardial cells decreased at day 1 and heart function improved at day 28, with increased neovascularization (P < 0.05). These results indicate that hPAMAM-based gene delivery into myoblasts is feasible and effective and may serve as a novel and promising non-viral DNA vehicle for gene therapy in myocardial infarction.
我们研究了超支化聚酰胺-胺（hPAMAM）介导的人血管内皮生长因子-165（hVEGF（165））基因转染骨骼肌成肌细胞用于心脏修复的可行性和疗效。hPAMAM 是通过改良的一锅法合成的。封装的 DNA 被 hPAMAM 保护，在 120 分钟以上的时间内不会降解。在最佳条件下，hPAMAM 在成肌细胞中的转染效率为 82.6±7.0%，细胞活力为 94.6±1.4%。与低细胞毒性的聚醚酰亚胺和 Lipofectamine 2000 相比，hPAMAM 显示出更高的转染效率（P<0.05）。转染的骨骼肌成肌细胞可稳定表达 hVEGF（165）达 18 天。移植 hPAMAM-hVEGF（165）转染细胞后，第 1 天凋亡的心肌细胞减少，第 28 天心脏功能改善，新生血管增加（P<0.05）。这些结果表明，hPAMAM 介导的成肌细胞基因转染是可行和有效的，可能成为心肌梗死基因治疗的一种新型有前途的非病毒 DNA 载体。
