正常和炎症来源的人牙周膜干细胞内皮向分化能力的比较

A comparative study on the endothelial cell differentiation of human healthy and inflammatory periodontal ligament stem cells

目的:比较炎症和正常牙周膜组来源的牙周膜干细胞(iPDLSCs和PDLSCs)体外成血管的能力。方法:组织块酶消化法培养iPDLSCs和PDLSCs,并经有限稀释法纯化和干细胞表面标记物检测鉴定后,对其进行成血管诱导,并通过免疫荧光染色、实时定量PCR、Matrigel assay检测其内皮细胞相关标记物的表达水平及毛细血管网形成状况。结果:两种细胞经纯化后均阳性表达干细胞标记物CD90、CD29、CD105、CD146;iPDLSCs比PDLCSs具有更高的克隆形成能力;两种细胞经内皮向诱导培养后其内皮细胞标记物VEGF、vWF、CD31、VE-cadherin mRNA的表达水平均较诱导前明显上调(P<0.05),并且均能在体外形成管腔样结构;iPDLSCs中CD31、VE-cadherin mRNA的表达水平和管腔长度、分支节点数等均明显高于PDLSCs(P<0.05)。结论:正常和炎症来源的人牙周膜干细胞均具有成血管能力,并且炎症来源的成血管能力更高。

AIM： To compare the angiogenesis ability of the stem cells derived from inflammatory and healthy periodontal ligament tissues （iPDLSCs and PDLSCs）in vitro. METHODS： iPDLSCs and PDLSCs were prima- rily cultured by tissue digestion method. Cells were passaged and identified by stem cell surface marker expression using flow cytometry. The angiogenesis ability of the cells were determined by angiogenesis induction, immunofluorescence test, real-time quantitative PCR and Matrigel assay. RESULTS ： The two types of cells both expressed stem cell markers CDg0, CD29, CD105 and CD146. iPDLSCs showed higher colony forming capacity. After angiogenesis induction, both cells could express endothelial cell markers VEGF, vWF, CD31, VE- cadherin and form tube structure in vitro. However, CD31 and VE - cadherin expression of iPDLSCs was higher than that of PDLSCs （P 〈 0.05）, the ves- sel length and branch points in the culture of iPDLSCs were more than those in the culture of PDLSCs. CONCLUSION： Human PDLSCs and iPDLSCs both have angiogenesis ability in vitro, and the ability of iPDLSCs is stronger than that of PDLSCs.