摘要/Abstract

摘要： 目的 ·探讨聚（左旋乳酸 -己内酯）［poly（L-lactic acid caprolactone），PLCL］/明胶静电纺丝支架对内皮祖细胞（ endothelial progenitor cells，EPCs）成血管分化的影响。方法 ·分离大鼠 EPCs后进行培养、鉴定。制作 PLCL/明胶静电纺丝支架，进行扫描电子显微镜及水接触角测试。 EPCs种植于 PLCL/明胶静电纺丝支架上， CCK8法检测细胞增殖。采用 RT-PCR检测成血管相关基因血管内皮生长因子（vascular endothelial growth factor，Vegf）、激酶插入区受体（kinases region receptor，Kdr）表达，Western blotting技术检测 VEGF蛋白表达。结果 ·密度梯度离心配合差速贴壁法可有效分离 EPCs，PLCL/明胶静电纺丝纳米纤维高密度多孔，亲水性能有利于细胞黏附， EPCs在支架上生长良好。 PLCL/明胶组 Vegf及 Kdr基因的表达较对照组显著增加（ P0.000），VEGF蛋白表达也显著增强（P0.000）。结论 · PLCL/明胶是组织工程理想支架，且对 EPCs成血管分化有促进作用。
关键词: 聚（左旋乳酸 -己内酯）, 内皮祖细胞, 分离培养, 生物学鉴定, 静电纺丝, 成血管
Abstract:
Objective · To investigate the effect of poly (L-lactic acid caprolactone) (PLCL) /gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs). Methods · Rat bone marrow-derived EPCs were isolated and cultured, then identification was performed. After preparation of PLCL/gelatin blend electrospun scaffold, scanning electron microscopy and water contact angle test were carried out. EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation. The of vascular endothelial growth factor (Vegf ) and kinases region receptor (Kdr) was observedRT-PCR and the of VEGF protein was observedWestern blotting. Results · The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs. PLCL/gelatin electrospun nanofibers were porous, and the hydrophilic properties were favorable for cell adhesion, and EPCs grew well on the scaffold. The of Vegf and Kdr gene in PLCL/gelatin group was higher than that in control group (P0.000), and the of VEGF protein was also increased (P0.000). Conclusion · PLCL/gelatin is an ideal scaffold for tissue engineering, and it can promote the angiogenesis differentiation of EPCs.
Key words: poly (L-lactic acid caprolactone), endothelial progenitor cell, isolation and culture, biological identification, electrospun, angiogenesis


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