氟暴露对人牙周膜干细胞增殖及成骨分化的影响  

Effects of Fluoride Exposure on Cell Proliferation and Osteogenic Differentiation of Periodontal Ligament Stem Cells

作　　者：邱吟枫 汤颖 沈忆芬 刘超 沈昊 顾永春 于金华 QIU Yinfeng;TANG Ying;SHEN Yifen;LIU Chao;SHEN Hao;GU Yongchun;YU Jinhua(Ninth People's Hospital of Suzhou, Suzhou 215200, China;Jiangsu Key Laboratory of Oral Diseases, Nanjing 210029, China)

机构地区：[1]苏州市第九人民医院口腔科,江苏苏州215200 [2]江苏省口腔疾病研究重点实验室,江苏南京210029

出　　处：《口腔医学研究》2020年第9期866-870,共5页Journal of Oral Science Research

基　　金：江苏省口腔疾病研究重点实验室开放课题基金(编号:JSK-LOD-KF-1705);苏州市吴江区卫健委科教兴卫项目(编号:wwk 201705)。

摘　　要：目的:探讨氟暴露对牙周膜干细胞(periodontal ligment stem cells,PDLSCs)增殖及成骨分化的影响。方法:从人牙周膜组织中分离、培养得到PDLSCs,取第3代细胞行流式细胞分析干细胞表面标志物。分别用含0.1、1、10、20、40 mg/L的含氟培养基进行培养,检测不同浓度氟对PDLSCs细胞增殖(CCK-8法)、集落形成、细胞周期变化(流式细胞分析)、成骨分化能力(茜素红染色及Western blot分析ALP及RUNX2的蛋白表达水平)的影响。结果:氟暴露对PDLSCs增殖与成骨分化的影响呈浓度及时间依赖性。0.1~10 mg/L氟对PDLSCs增殖无显著影响(P>0.05),但0.1及1 mg/L氟可显著促进其成骨分化,表现为钙化结节增多(P<0.05),ALP及RUNX2蛋白表达水平升高(P<0.05)。高浓度氟(20、40 mg/L)显著抑制PDLSCs增殖(P<0.05),具有较强的细胞毒性。结论:氟暴露会影响PDLSCs的增殖与成骨分化。高浓度氟抑制PDLSCs增殖,而氟浓度较低时(0.1、1 mg/L)对PDLSCs增殖无显著影响,并可以促进细胞成骨分化。Objective:To disclose the biological effects of fluoride(F)exposure on cell proliferation and osteogenic differentiation of periodontal ligament stem cells(PDLSCs).Methods:PDLSCs were isolated and cultured from human periodontal ligament tissues.The surface markers of passage 3 cells were analyzed by flow cytometry.After treated with different concentrations of F(0.1-40 mg/L)for indicated period of time,the effects of F exposure on cell proliferation(CCK-8 assay),capability of colony-forming,cell circle phase changes(flow cytometry),as well as the osteogenic potential(Alizarin red staining and Western blot assay of expression of RUNX2 and ALP)were examined.Results:F exposure affected the cell proliferation and osteogenic differentiation of PDLSCs in a dose-and time-dependent manner.0.1-10 mg/L F did not significantly affect cell proliferation(P>0.05),while 0.1 and 1 mg/L F significantly enhanced the osteogenic potential,which manifested as increased calcified nodules and upregualtion of RUNX2 and ALP expression(P<0.05).High dose of F(20 and 40 mg/L)significantly inhibited the cell proliferation(P<0.05),exhibiting remarkable cytotoxicity.Conclusion:F exposure affects the proliferation and osteogenic differentiation of PDLSCs.High concentration of F inhibits the proliferation of PDLSCs,while low F concentration(0.1 and 1 mg/L)has no significant effect on the proliferation of PDLSCs,and can promote cell osteogenic differentiation.

关 键 词：牙周膜间充质干细胞 氟暴露 细胞增殖 细胞分化 

分 类 号：R73[医药卫生—肿瘤]