不同浓度雌激素对人源性牙周膜干细胞成骨分化能力的影响

摘要

Objective:To compare the effect of estrogen with different concentrations on osteogenic differentiation of human periodontal ligament stem cells. Methods: Human periodontal ligament stem cell suspension was prepared with DMEM complete medium and then were cocultured with 0,10í10-10,10í10-9,10í10-8,10í10-7,10í10-6,and 10í10-5 mol/L of estrogen. Real-time quantitative PCR was used to detect the expression of osteogenic key indicators like Runx2,ALP,and OCN mRNA after coculture. Results:No toxic effect on human periodontal ligament stem cells was found in 10í10-10,10í10-9,10í10-8,10í10-7,10í10-6,and 10í10-5 mol/L estrogen groups and no statistically sig-nificant difference was noted among these groups (P>0.05). On day 7,14,and 21 during osteogenic differentiation, the expression of Runx2,ALP,and OCN mRNA were the highest in 10í10-7 estrogen group(P<0.05~0.01) and were higher in all estrogen groups compared with the negative control group (0 estrogen group,P<0.05). Conclusion:Es-trogen can promote bone-related gene expressions during osteogenic differentiation of human periodontal ligament stem cells in a dose-dependent manner when its concentration was not more than 10í10-7mol/L. When estrogen levels were more than 10í10-7mol/L,the effect was not seen.%目的：对比研究不同浓度的雌激素对人源性牙周膜干细胞成骨分化的影响。方法：采用DMEM完全培养基将人牙周膜干细胞制成单细胞悬液，分别加入浓度为0、10×10-10、10×10-9、10×10-8、10×10-7、10×10-6、10×10-5mol/L的雌激素，以未加入雌激素的成骨诱导液组（普通DMEM培养基中加入10mMβ-甘油磷酸钠、50μM维生素C及10nM地塞米松）作为阴性对照。培养后实时定量PCR方法分别检测成骨早中晚期关键指标Runx2、ALP和OCN mRNA表达。结果：浓度为10×10-10、10×10-9、10×10-8、10×10-7、10×10-6、10×10-5mol/L的雌激素对人源性的牙周膜干细胞培养均无毒性；不同浓度组间差异无统计学意义（P＞0.05）；在成骨诱导7、14、21天时，Runx2、ALP及OCN mRNA表达在10×10-7组最高（P＜0.05~0.01）。在不同浓度雌激素组中成骨相关基因的表达均高于阴性对照组（P＜0.05）。结论：当雌激素浓度＜10×10-7mol/L时，雌激素能够以剂量依赖的方式促进人牙周膜干细胞成骨分化过程中成骨相关基因的表达。当雌激素浓度＞10×10-7mol/L时，雌激素浓度的增加并未促进成骨相关基因的表达。