Transplantation of nanoparticle transfected skeletal myoblasts overexpressing vascular endothelial growth factor-165 for cardiac repair
Transplantation of nanoparticle transfected skeletal myoblasts overexpressing vascular endothelial growth factor-165 for cardiac repair
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DOI：
10.1161/circulationaha.106.680124
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发表时间：
2007-09-11
期刊：
影响因子：
37.8
通讯作者：
Sim, Eugene K. W.
中科院分区：
文献类型：
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作者：
Ye, Lei;Haider, Husnain Kh;Sim, Eugene K. W.
背景-我们研究了基于聚乙烯亚胺（PEI）的人血管内皮生长因子-165的可行性和有效性（hVEGF（165））基因转移到人骨骼肌成肌细胞（HSM）中用于基于细胞的递送到梗塞心肌。方法和结果-基于使用增强型绿色荧光蛋白（pEGFP）的优化转染程序，用PEI（PEI-phVEGF（165））纳米颗粒携带的质粒-hVEGF（165）（phVEGF（165））转染HSM。将转染hVEGF（165）的HSM移植到急性心肌梗死（AMI）大鼠心脏模型中，观察hVEGF（165）的转染和表达情况：第1组为DMEM注射组，第2组为HSM移植组，第3组为PEI-phVEGF（165）转染的HSM（PEI-phVEGF（165）myoblast）移植组。共48只大鼠从细胞移植前3天至移植后4周接受环孢素注射。进行超声心动图以评估心脏功能。处理后4周处死动物，对心脏组织进行分子和组织学研究。基于优化的转染条件，转染的HSM表达hVEGF（165）18天，体外细胞存活率>90%。与组1相比，组2和组3的凋亡指数降低。在第4周，与第1组和第2组相比，第3组中PECAM-1免疫染色的血管密度（x400）显著更高（两者均为P = 0.043）。与组1和组2相比，组3中左心室前壁的局部血流量（ml/min/g）较高（两组均为P = 0.043）。结论PEI纳米粒介导的hVEGF（165）基因转染HSM是安全可行的。它可能作为一种新的和有效的替代血管平滑肌在心脏修复。
Background - We investigated the feasibility and efficacy of polyethylenimine (PEI) based human vascular endothelial growth factor-165 (hVEGF(165)) gene transfer into human skeletal myoblasts (HSM) for cell based delivery to the infarcted myocardium.Methods and Results - Based on optimized transfection procedure using enhanced green fluorescent protein (pEGFP), HSM were transfected with plasmid-hVEGF(165) (phVEGF(165)) carried by PEI (PEI-phVEGF(165)) nanoparticles. The transfected HSM were characterized for transfection and expression of hVEGF(165) in vitro and transplanted into rat heart model of acute myocardial infarction (AMI): group-1 = DMEM injection, group-2 = HSM transplantation, group-3 = PEI-phVEGF(165)-transfected HSM (PEI-phVEGF(165) myoblast) transplantation. A total of 48 rats received cyclosporine injection from 3 days before and until 4 weeks after cell transplantation. Echocardiography was performed to assess the heart function. Animals were sacrificed for molecular and histological studies on the heart tissue at 4 weeks after treatment. Based on optimized transfection conditions, transfected HSM expressed hVEGF(165) for 18 days with >90% cell viability in vitro. Apoptotic index was reduced in group-2 and group-3 as compared with group-1. Blood vessel density (x400) by immunostaining for PECAM-1 in group-3 was significantly higher (P = 0.043 for both) as compared with group-1 and group-2 at 4 weeks. Regional blood flow (ml/min/g) in the left ventricular anterior wall was higher in group-3 (P = 0.043 for both) as compared with group-1 and group-2. Improved ejection fraction was achieved in group-3 (58.44 +/- 4.92%) as compared with group-1 (P = 0.004).Conclusion - PEI nanoparticle mediated hVEGF(165) gene transfer into HSM is feasible and safe. It may serve as a novel and efficient alternative for angiomyogenesis in cardiac repair.