人牙周膜细胞在矿化液作用下的增殖、成骨分化与相关基因表达

Cell Proliferation,Osteogenic Differentiation,and Relative Gene Expression of Human Periodontal Ligament Cells Affected by Mineralizaion Medium.

目的:分离培养人牙周膜细胞(PDLCs),观察矿化液对PDLCs细胞增殖和成骨分化的影响。方法:酶解组织块后获得人PDLCs,矿化液处理后观察对细胞影响。采用MTT法检测对细胞增殖;流式细胞分析细胞周期改变;分别通过碱性磷酸酶(ALP)染色、茜素红染色,RT-PCR观察矿化液诱导成骨分化的效果。结果:矿化液对人PDLCs的增殖无显著影响,高浓度地塞米松(Dex)的矿化液有一定抑制作用。含Dex的矿化液可促进成骨分化,包括ALP活性提高,矿化结节形成,以及成骨基因的表达上调。S100A4和PLAP-1是相对特异的牙周膜标志,也参与矿化液诱导的成骨分化的调控过程。结论:含地塞米松的矿化液可有效促进人PDLCs的成骨分化,对细胞增殖影响较小。PDLCs是一种新型的种子细胞,可应用于骨组织工程。

Objecticve： To isolate human periodontal ligament cells （PDLCs）, and determine the effects of miner- alizaion medium on proliferation and osteogenic differentiation of PDLCs. Methods： PDLCs were purified from ex- tracted teeth using enzyme digestion, and mineralizaion medium was prepared to determine its effect on PDLCs. Cell proliferation was examined by MTT, and cell cycle analysis was performed using flow cytometry. Osteogenic differentiation was assessed by alkaline phosphatase activity （ALP）, Alizarin red staining, and RT--PCR. Results： Mineralizaion medium was not cytotoxic to PDLCs, although high dose of dexamethasone （Dex） could slightly in- hibit cell proliferation. Mineralizaion medium containing Dex promoted osteogenic differentiation of PDLCs in a dose --dependent manner, demonstrating increasing ALP activity, mineral deposition, and related osteogenic--gene ex- pression. PDL markers $100A4 and PLAP--1 were involved in the regulation of mineralizaion medium induced os- teogenic differentiation. Conclusion： The present study demonstrates that mineralizaion medium containing Dex pro- motes osteogenic differentiation of PDLCs effectively, but not cytotoxic, which is promising for future application of PDLCs in bone tissue engineering.