摘要
背景:钙离子作为第二信使,调控着众多的生理过程,但在牙周膜干细胞成骨分化方面的生物学效应尚不明确。目的:探讨钙离子在人牙周膜干细胞增殖和成骨分化过程中的作用。方法:使用含钙离子浓度为0(对照组),1,2,5,10,15 mmol/L的完全培养基及成骨诱导液对第3代人牙周膜干细胞进行增殖能力检测及成骨诱导。培养1,3,5,7 d使用CCK-8检测细胞增殖能力;在成骨诱导第3,7天采用Real-Time PCR和Western blot检测RUNX2、OPN、OCN的基因及蛋白表达;成骨诱导第14天时进行茜素红染色分析人牙周膜干细胞的成骨矿化情况。结果与结论:①CCK-8检测结果显示,2,5 mmol/L钙离子组人牙周膜干细胞在第5,7天的增殖能力优于对照组(P<0.05);②体外成骨诱导7 d后,5,10 mmol/L钙离子组人牙周膜干细胞中RUNX2、OPN、OCN的表达高于对照组(P<0.05);③体外成骨诱导14 d后,含钙离子组人牙周膜干细胞的矿化结节数量均高于对照组(P<0.05);④以上结果表明,在一定浓度范围内,钙离子可促进人牙周膜干细胞的增殖与成骨分化。
BACKGROUND:As a second messenger,Ca^(2+) regulates many physiological processes,but its biological effect on the osteogenic differentiation of periodontal ligament stem cells is still unclear.OBJECTIVE:To investigate the effect of Ca^(2+) on the osteogenic differentiation of human periodontal ligament stem cells.METHODS:The proliferation ability and osteogenic differentiation of the third generation human periodontal ligament stem cells were detected by the complete medium containing Ca^(2+) at the concentrations of 0(control group),1,2,5,10,and 15 mmol/L and osteogenic induction solution.CCK-8 assay was used to detect cell proliferation at 1,3,5,and 7 days.Real-time PCR and western blot assay were used to detect the expression levels of genes and proteins related to osteogenic differentiation factors RUNX2,OPN and OCN on days 3 and 7 of osteogenic induction.Alizarin red staining was performed on day 14 to analyze the osteogenic mineralization of human periodontal ligament stem cells.RESULTS AND CONCLUSION:(1)CCK-8 assay showed that the proliferation ability of human periodontal ligament stem cells in 2 and 5 mmol/L groups was better than that in control group on days 5 and day 7(P<0.05).(2)On day 7 after in vitro osteogenic induction,the expression levels of osteogenic genes RUNX2,OPN and OCN in human periodontal ligament stem cells of 5 and 10 mmol/L groups were higher than those in control group(P<0.05).(3)After 14 days of in vitro osteogenic induction,the numbers of mineralization nodules of human periodontal ligament stem cells in Ca^(2+) groups were higher than that in control group(P<0.05).(4)These results suggest that Ca^(2+) can promote the proliferation and osteogenic differentiation of human periodontal ligament stem cells in a certain concentration range.
作者
李颖辉
齐芳芳
韩行
赵力如
马文盛
Li Yinghui;Qi Fangfang;Han Xing;Zhao Liru;Ma Wensheng(Department of Orthodontics,Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050000,Hebei Province,China;Hebei Key Laboratory of Stomatology,Shijiazhuang 050000,Hebei Province,China;Department of Operative Dentistry and Endodontics,Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050000,Hebei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第19期3005-3010,共6页
Chinese Journal of Tissue Engineering Research
基金
河北省卫生健康委医学科学研究课题计划(20191077),项目负责人:李颖辉
河北省人社厅引进留学人员资助项目(C201858),项目负责人:李颖辉。
关键词
钙离子
人牙周膜干细胞
增殖
成骨分化
Ca2+
human periodontal ligament stem cell
proliferation
osteogenic differentiation
