外源性雌激素对人牙周膜干细胞骨分化能力影响的实验研究

Experimental study on the effects of exogenetic estrogen on the ability of human periodontal ligament stem cell osteogenic differentiation

目的检测外源性雌激素对人牙周膜干细胞(PDLSCs)骨分化能力的影响。方法将体外培养的PDLSCs加入无酚红成骨诱导培养液和不同浓度的17β-E2[分为E10^(-7)、E10^(-8)、E10^(-9)组和对照组(成骨诱导培养液+无水乙醇(0.01%)]-雌二醇[分为E10^(-7)、E10^(-8)、E10^(-9)组、ICI组(成骨诱导培养液+ICI182780)、E10^(-7)+ICI组(成骨诱导培养液+1×10^(-7)17β-E2+1×10^(-7)ICI182780)],观察各组细胞形态、测定其增值水平、碱性磷酸酶(ALP)活性、Ⅰ型胶原合成能力。结果添加外源性雌激素后,细胞增殖加快,生长密集,呈螺旋状排列。PDLSCs的增殖发生变化,与对照组相比,第3、5天雌激素干扰组细胞增殖受到抑制(P<0.05),此后雌激素干扰组细胞增殖高于对照组(P<0.05),在不同药物浓度组间,E10^(-7)组对细胞增殖影响最为明显,其细胞增殖水平高于E10^(-8)、E10^(-9)组(P<0.05)。ALP表达显示动态变化,自第3天开始,各组ALP表达均升高,而雌激素干扰组ALP表达量高于对照组(P<0.05),在不同药物浓度组间E10^(-7)组ALP表达增加高于E10^(-8)、E10^(-9)组(P<0.05),与ICI组、E10^(-7)+ICI组比较,差异无统计学意义(P>0.05);PDLSCsⅠ型胶原合成表达与对照组相比有增加趋势,且与药物浓度有剂量依赖关系。结论雌激素对PDLSCs成骨分化过程有促进作用,该作用与雌激素浓度密切相关。

Objective To investigate the effects of exogenetic estrogen on the differentiation ability of human periodontal ligament stem cell（ PDLSCs） in vitro. Methods The PDLSCs were cultured in non-phenol red osteogenesis induction medium with different concentrations of 17β-E2（ E10^（-7）,E10^（-8,）E10^- 9））,moreover,which were cultured in osteogenesis induction medium ＋ 0. 01% dehydrated alcohol（ control group）,besides,-which were cultured in the medium with different concentrations of estradiol（ E10^（-7）,E10^（-8）,E10^（-9）） and were cultured in osteogenesis induction medium ＋ICI182780（ ICI group）,and osteogenesis induction medium ＋ 10^（-7）β-E2 ＋ 10^（-7） ICI182780（ 10^（-7）＋ ICI group）,respectively.The cell shape,cell proliferation activity,alkaline phosphatase（ ALP） activity,the synthesis ability of type Ⅰ collagen were detected in every group. Results After treated by exogenetic estrogen,the cell proliferation was speeded up,cell growth was intensived,with spiral arrangement. As compared with that in control group,the cell proliferation on 3 days,5 days in estrogen intervention group was obviously inhibited（ P 0. 05）,moreover,the cell proliferation in estrogen intervention group was higher than that in control group（ P 0. 05）. Among difeerent drug concentration groups,the effects in E10^（-7） group on cell proliferation were the most obvious,and the cell proliferation levels in E10^（-7） group were significantly higher than those in E10^（-8） group and E10^（-9） group（ P 0. 05）. The espression of ALP showed dynamic changes,the levels of ALP from the third day in all the groups were increased,however,which in estrogen intervention group were significantly higher than those in control group（ P 0. 05）. Among different drug concentration groups,the levels of ALP in E10^（-7） group were significantly higher than those in E10^（-8） group and in E10^（-9） group（ P 0. 05）. However there were no significant differences between ICI group and E10^（-7） ＋ ICI group（ P 0. 05）. As compared with that in control group,the type Ⅰ collagen synthesis of PDLSCs had an enhancement trend,moreover,with a drug dose-dependent way. Conclusion The estrogen can promote osteogenic differentiation process in PDLSCs,moreover,its action is closely correlated to estrogen concentration.