Novel hyperbranched polyamidoamine nanoparticles for transfecting skeletal myoblasts with vascular endothelial growth factor gene for cardiac repair
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We investigated the feasibility and efficacy of hyperbranched polyamidoamine (hPAMAM) mediated human vascular endothelial growth factor-165 (hVEGF165) 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 hVEGF165 expression for 18 days. After transplantation of hPAMAM–hVEGF165 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.
KeywordsVascular Endothelial Growth Factor Left Ventricular Ejection Fraction Reverse Transcription Polymerase Chain Reaction Transfection Efficiency Vascular Endothelial Growth Factor Expression
We are grateful for the support of Shanghai Pujiang Program (Grant No. 10PJ1402000), the Doctor Project for Young Teachers of Ministry of Education (Grant No. 20090071120032), and the National Science Foundation of China (Grant No. 20874015).
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