基质细胞衍生因子-1对炎症和正常来源的人牙周膜干细胞成骨分化能力的影响

Comparative study with the effect of stromal cell derived factor-1 on osteogenic differentiation of human healthy and inflammatory periodontal ligament stem cells

目的通过体外培养炎症来源的人牙周膜干细胞(iPDLSCs)和正常来源的人牙周膜干细胞(hPDLSCs),比较基质细胞衍生因子-1(SDF-1)对于两种来源细胞的成骨分化作用。方法采用组织块酶消化法原代培养iPDLSCs和hPDLSCs,经有限稀释法纯化,通过流式细胞仪对干细胞表面标记物检测鉴定后,对其进行成骨诱导;MTT法检测并比较SDF-1对两种来源的细胞增殖能力的影响;茜素红染色检测SDF-1作用于两种来源的细胞后钙化骨量的表达;碱性磷酸酶法比较SDF-1作用于两者的成骨分化能力;逆转录聚合酶链反应(RT-PCR)法检测SDF-1作用于两种牙周膜干细胞前后成骨相关基因表达水平的变化。结果两种来源的牙周膜细胞经纯化后均阳性表达干细胞标记物。hPDLSCs较iPDLSCs增殖能力高;两种细胞经SDF-1成骨诱导培养后,成骨相关基因的表达水平均较诱导前明显上调(P<0.05),SDF-1在50、200ng·mL^-1时分别对iPDLSCs和hPDLSCs细胞成骨分化作用最明显(P<0.05)。结论正常来源和炎症来源的人牙周膜干细胞均具有成骨分化能力,SDF-1可增强两种来源的牙周膜干细胞的成骨分化能力。

Objective This study aims to compare the osteogenic differentiation capability of stem cells derived from human inflammatory periodontal ligament tissues (iPDLSCs) with those of stem cells derived from healthy periodontal ligament tissues (hPDLSCs). Both types of tissues were induced by stromal cell derived factor (SDF-1) in vitro. Methods iPDLSCs and hPDLSCs were primarily cultured by tissue digestion method and purified by limited dilution cloning. The cells were passaged and identified by stem cell surface marker expression through flow cytometry. Then, we used thiazolyl blue tetrazolium brom ide to detect and compare the proliferation capabilities of the iPDLSCs and hPDLSCs. Express of bone volumes were detected by alizarin red staining after SDF-1 was added to the cells. Using alkaline phosphatase, we evaluated the osteogenic differentiation capability of the cells induced by SDF-1. The expression levels of the osteogenesis-related genes of the cells induced by SDF-1 were determined by reverse transcription-polymerase chain reaction. Results After purification, both iPDLSCs and hPDLSCs expressed stem cell markers. hPDLCSs had a higher proliferation capability than iPDLSCs. Osteogenesisrelated genes had higher expression levels in the cells induced by SDF-1 than in those without induction (P<0.05). SDF-1 at 50 and 200 ng·mL^-1 concentration greatly affected the differentiation capabilities of iPDLSCs and hPDLSCs respectively. Conclusion iPDLSCs and hPDLSCs had osteogenic differentiation capability. The level of osteogenic differentiation in normal and inflamed periodontal ligament stem cells increases after SDF-1 induction.