不同浓度血管内皮细胞生长因子对人牙周膜干细胞内皮向分化的影响

›› 2020, Vol. 40 ›› Issue (6): 506-512.

不同浓度血管内皮细胞生长因子对人牙周膜干细胞内皮向分化的影响

石笑¹,赵寅华²,赵萤²,程百祥¹,陈永进³,张旻²

1. 空军军医大学第三附属口腔医院
2. 第四军医大学口腔医院
3. 第四军医大学口腔医学院急诊与综合临床科

收稿日期:2019-12-10 修回日期:2020-01-21 出版日期:2020-06-28 发布日期:2020-06-28
通讯作者: 张旻 E-mail:zhangmin@fmmu.edu.cn
基金资助:
国家自然科学基金;国家自然科学基金

Effects of vascular endothelial growth factor of different concentrations on endothelial differentiation of human periodontal ligament stem cells.

Received:2019-12-10 Revised:2020-01-21 Online:2020-06-28 Published:2020-06-28
Contact: ZHANG MIN E-mail:zhangmin@fmmu.edu.cn

摘要/Abstract

摘要： 【摘要】目的：探讨不同浓度血管内皮细胞生长因子（VEGF）诱导牙周膜干细胞（PDLSCs）血管内皮向分化的作用。方法：体外分离培养的PDLSCs用bFGF（10 ng/mL），IGF-1（2 ng/mL）及不同浓度的VEGF（10, 20, 50 ng/mL）诱导。7 d、14 d和21 d后，倒置相差显微镜观察细胞形态学变化，管腔形成实验检测细胞的管腔形成能力，细胞免疫荧光法检测vWF、VEGFR2的表达水平，western blotting法检测细胞中血管内皮特异性指标ANG2、VEGFR-2、VEGFA、IGF-1及vimentin的表达水平。结果：诱导后的PDLSCs形态明显改变，管腔形成实验显示，细胞互相连接，形成不同数量的管腔样结构，细胞免疫荧光染色显示，vWF和VEGFR-2表达阳性，western blotting结果显示，ANG2、VEGFR-2、VEGFA、IGF-1呈阳性表达，vimentin表达阴性，并且以高浓度VEGF组诱导14 d和21 d最为显著。结论：PDLSCs具有向内皮细胞分化的潜能，且以含10 ng/mL bFGF，2 ng/mL IGF-1及50 ng/mL VEGF的内皮分化诱导液较为理想，诱导时间以14 d和21 d为宜。

关键词: 【关键词】牙周膜干细胞, 血管内皮细胞生长因子, 血管内皮细胞, 诱导分化

Abstract: Abstract: Objective To investigate the effects of vascular endothelial growth factor (VEGF) on endothelial differentiation of periodontal ligament stem cells (PDLSCs). Methods PDLSCs were isolated and induced by bFGF (10 ng/mL), IGF-1 (2 ng/mL) and VEGF of different concentrations (10, 20, 50 ng/mL) in vitro. After 7 d, 14 d and 21 d of induction, the morphology of PDLSCs were observed by inverted microscope. The lumen forming ability of PDLSCs were measured by the tube formation assay. The expression levels of vWF and VEGFR-2 were detected by immunofluorescence. The expression of ANG2, VEGFR-2, VEGFA, IGF-1 and vimentin were detected by western blotting. Results After induction, the morphology of PDLSCs was obviously changed, tube formation assay showed PDLSCs all connected with each other to form different numbers of lumen like structures. Immunofluorescence results showed positive for vWF and VEGFR-2, western blotting results showed ANG2, VEGFR-2, VEGFA and IGF-1 were positively expressed, while vimentin was negatively expressed, and the high-concentration VEGF group was most obvious at days 14 d and 21 d post-induction. Conclusion?PDLSCs possessed the potential for differentiation into endothelia like cells, and all results suggested that 50 ng/mL VEGF was an optimum induction concentration and the optimum duration of induction were 14 d and 21 d.

Key words: [keywords] periodontal ligament stem cells, vascular endothelial growth factor, vascular endothelial cells, induced differentiation.

中图分类号:

R349.5

石笑赵寅华赵萤程百祥陈永进张旻. 不同浓度血管内皮细胞生长因子对人牙周膜干细胞内皮向分化的影响[J]. 口腔医学, 2020, 40(6): 506-512.

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