制备可缓释血管内皮生长因子的聚乳酸-聚乙醇酸支架及其与血管平滑肌细胞的生物相容性

Preparation of poly (DL-lactic-co-glycolic acid) scaffold for sustained release of vascular endothelial growth factor and its biocompatibility with vascular smooth muscle cells

目的:制备一种新型的可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架,探讨其与血管平滑肌细胞的细胞相容性。方法:实验于2006-12/2007-05在解放军第四军医大学西京医院心内科实验室(省部级重点实验室)完成。以明胶微球作为缓释载体,加载血管内皮生长因子和聚乳酸-聚乙醇酸复合构建组织工程血管支架并考察体外释放效果。采用体外培养的兔血管平滑肌细胞种植在可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架膜片上,用相差显微镜观察细胞的生长情况,绘制细胞生长曲线,并用MTT法测定细胞增殖情况。实验过程中对动物处置符合动物伦理学要求。结果:血管内皮生长因子在体外释放14d以上,血管平滑肌细胞在可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架膜片上生长良好;MTT法检测细胞增殖情况显示,可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架组平滑肌细胞增殖较快,细胞较为活跃(P<0.05)。结论:可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架制备方法简便易行,生长因子活性得以保存,对血管平滑肌细胞具有良好的相容性,可以用于组织工程血管的进一步构建。

AIM： To prepare a biodegradable poly（DL-lactic-co-glycolic acid） （PLGA） scaffold with vascular endothelial growth factor （VEGF） sustained-release system and explore its biocompatibility with vascular smooth muscle cells （SMC）. METHODS： The experiment was carried out in the Laboratory of Angiocardiopathy （Key Laboratory of Provincial Level）, Xijing Hospital, the Fourth Military Medical University of Chinese PLA from December 2006 to May 2007. VEGF incorporated into the gelatin microspheres was combined with PLGA scaffold and the release efficiency in vitro was observed. The rabbit vascular SMC cultured in vitro were inoculated onto VEGF-PLGA membrane, their growth was observed by phase, contrast microscopy, cell growth curve was drawn, and proliferation of cells was determined by MTT method. All the experimental procedures met the ethnical requirement of animals. RESULTS： VEGF was released in vitro for over 14 days, vascular SMC grew well on VEGF-PLGA scaffold and showed significant better proliferation than other groups by MTT （P 〈 0.05）. CONCLUSION： The preparative method of biodegradable VEGF-PLGA scaffold is convenient and benefits for releasing VEGF, it has well compatibility with rabbit vascular SMC, thus VEGF-PLGA scaffold might be a promising tissue engineering vascular scaffold.