聚四氟乙烯血管材料携带的血管内皮生长因子基因转染血管内皮细胞并促进其生长的可行性

Endothelial cell proliferation on polytetrafluoroethylene vascular graft materials containing VEGF gene plasmid

目的探讨聚四氟乙烯(PTFE)血管材料携载血管内皮生长因子(VEGF)基因转染内皮细胞并促进其生长的可行性。方法将pCDI-hVEGF_(121)/pCDI/pEGFP．质粒溶液处理过的PTFE人工血管修剪成圆片,置于生理盐水中并于不同的时间测溶液中DNA浓度,绘出体外释放曲线。分离和培养人脐静脉内皮细胞,将内皮细胞种植到携带上述质粒的PTFE材料表面,监测细胞增殖情况并用酶联免疫吸附试验(ELISA)法检测培养液中VEGF表达量,荧光显微镜下观察表达绿色荧光蛋白的内皮细胞比率。结果体外释放曲线提示从0．5～4．0 h的这段时间内质粒释放较快,随后减缓,直至72 h仍有质粒的释放。在24、72、120 h,pCDI-hVEGF_(121)组PTFE材料上的内皮细胞数和增长倍率都明显高于pCDI/pEGFP组材料(P＜0．05),pCDI-hVEGF_(121)组培养液的VEGF水平明显高于p CDI/pEGFP组(P＜0．01),荧光显微镜下观察到pEGFP组内皮细胞内绿色荧光蛋白的表达。结论证明了PTFE材料能够携带VEGF基因转染内皮细胞并促进其生长。

Objective To investigate whether vascular endothelial growth factor （VEGF） gene p[asmid carryiing polytetrafluoroethylene （PTFE） vascular graft material could transfect endothelial ceils （EC） and promote ECs growth. Methods The PTFE vascular graft materialscarrying pCDI-hVEGF121, pCDI or pEGFP plasmids were soaked in saline and the values of A260nm at different time points were measured, then the controlled release curve was made. ECs were derived from human umbilical vein and seeded on the PTFE materials carrying pCDI-hVEGF121, pCDI or pEGFP. The number of ECs was counted and VEGF protein concentrations were measured by ELISA method at different time points. The GFP expression in ECs on the PTFE materials carrying pEGFP was examined under the fluorescence microscopy. Results The controlled release curve showed the gene was released quickly during 0.5-4.0 h, then became slow,and released continuously even after 72 h. At 24,72 and 120 h, the number of ECs and proliferation rate on PTFE materials carrying pCDI-hVEGF121 were higher than that of PTFE materials carrying pCDI or pEGFP （P 〈 0.05）. The VEGF protein concentration in PTFE materials carrying pCDI-hVEGF121 was higher than that of PTFE materials carrying pCDI or pEGFP at 6,24,72 and 120 h （ P 〈 0.01 ）. The GFP expression in ECs on the PTFE materials carrying pEGFP could be detected under the fluorescence microscopy. Conclusion The PTFE graft can be used as carrier of VEGF gene plasmid. VEGF gene can transfect ECs and promote ECs growth.