人牙周膜干细胞成骨相诱导后相关基因的表达变化

目的:对人牙周膜干细胞进行筛选纯化,骨向诱导后观察其成骨能力,并对成骨相分化过程中相关基因的表达变化进行检测。方法:用组织块联合磁珠分选法分离纯化人牙周膜干细胞,初步探讨其成骨性能,碱性磷酸酶(ALP)和茜素红染色观察其成骨情况,实时定量PCR检测其相关成骨基因的表达变化。利用SPSS12.0软件包对数据进行统计学分析。结果:组织块联合磁珠分选法能成功培养出高纯度的人牙周膜干细胞。体外培养中,PDLSCs具备成脂和成骨的双相分化能力。在成骨诱导过程中,FOXO1和RUNX2的表达先升高再降低,ALP和OCN的基因表达水平持续增高。结论:成骨相关基因 ALP、RUNX2、FOXO1和OCN均参与其向成骨样组织分化的过程,其表达变化具有时间规律性,与成骨过程类似。

Osteogenic gene expression of human periodontal ligament stem cells during osteogenic induction

PURPOSE: To isolate and identify the human periodontal ligament stem cells, evaluate osteogenetic capacity, and investigate the changes of osteogenic bone related gene expression in mineralized medium at different times. METHODS: PDLSCs were isolated by tissue culture and magnetic activated cell sorting. Immunofluorescence staining was used for identification. The general situation of osteogenesis was assessed with alkaline phosphatase (ALP) and alizarin red staining. Real-time PCR was used to detect the expression of genes in osteoinduction. SPSS12.0 software package was used for data analysis. RESULTS: Tissue culture with magnetic cell sorting could isolate high-purity human periodontal ligament stem cells. During the osteogenic process, the expression of FoxO1 and Runx2 firstly increased and then decreased, ALP and OCN gene levels continued to increase. CONCLUSIONS: Similar to osteogenesis, ALP, Runx2, FoxO1 and OCN are regularly expressed during osteogenic induction.