上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (5): 499-.doi: 10.3969/j.issn.1674-8115.2018.05.003

• 论著·基础研究 • 上一篇    下一篇

聚 (左旋乳酸 -己内酯 )/明胶静电纺丝支架对大鼠骨髓 来源内皮祖细胞成血管分化的影响

张夏，郑雷蕾，刘艳，明叶，何昊珏，胡赟   

  1. 重庆医科大学附属口腔医院，口腔疾病与生物医学重庆市重点实验室，重庆市高校市级口腔生物医学工程重点实验室，重庆 401147
  • 出版日期:2018-05-28 发布日期:2018-05-28
  • 通讯作者: 胡赟，电子信箱：Huyuncqmu@163.com。
  • 作者简介:张夏（1993—），女，硕士生；电子信箱： 522414105@qq.com。
  • 基金资助:
    国家自然科学基金（81470772）；重庆市医学科研项目（20141013, 2015HBRC009）；重庆市自然科学基金（cstc2015jcyj10028, cstc2016jcyjA0238）

Effect of poly (L-lactic acid caprolactone)/gelatin blend electrospun on angiogenesis of rat bone marrow-derived endothelial progenitor cells

ZHANG Xia, ZHENG Lei-lei, LIU Yan, MING Ye, HE Hao-jue, HU Yun   

  1. Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Affiliated Stomatological Hospital, Chongqing Medical University, Chongqing 401147, China
  • Online:2018-05-28 Published:2018-05-28
  • Supported by:
    National Natural Science Foundation of China, 81470772; Chongqing Medical Research Project, 20141013, 2015HBRC009; Chongqing Natural Science Foundation, cstc2015jcyj10028, cstc2016jcyjA0238

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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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