人生长激素对人牙周膜干细胞增殖及成骨分化的影响  

Effects of human growth hormone on proliferation and osteogenic differentiation of human periodontal ligament stem cells

作　　者：杨俊辉 罗金莉 袁小平 Yang Junhui;Luo Jinli;Yuan Xiaoping(Department of Orthodontics of Stomatological Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Laboratory of Oral and Maxillofacial Reconstruction and Regeneration of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属口腔医院正畸科,四川省泸州市646000 [2]西南医科大学口颌面修复重建与再生实验室,四川省泸州市646000

出　　处：《中国组织工程研究》2021年第25期3956-3961,共6页Chinese Journal of Tissue Engineering Research

基　　金：泸州市政府-四川医科大学联合项目[2015LZCYD-S05(1/12)],项目负责人:袁小平。

摘　　要：背景:生长激素已被证明对于非牙源性间充质干细胞的增殖和骨向分化有促进作用,但对于人牙周膜干细胞的生物学效应还不明确。目的:探讨人生长激素对人牙周膜干细胞增殖以及成骨分化的影响。方法:用含0(对照组),10,100,200μg/L人生长激素的α-MEM完全培养基干预第3代人牙周膜干细胞,分别在第1,3,5,7天采用CCK-8法检测各组细胞增殖情况;用含0(对照组),10,100,200μg/L人生长激素的成骨诱导液干预第3代人牙周膜干细胞,第7天时采用RealTime PCR和Western blot检测骨向分化相关因子opn、runx2的基因和蛋白表达,第14天时进行茜素红染色以及茜素红半定量分析人牙周膜干细胞成骨后期矿化情况。结果与结论:100,200μg/L人生长激素组的人牙周膜干细胞增殖能力明显优于对照组(P <0.05);100,200μg/L人生长激素组opn、runx2的基因和蛋白表达高于对照组(P <0.05);100,200μg/L人生长激素组人牙周膜干细胞的矿化能力优于对照组(P <0.05);结果显示,在体外实验中,100,200μg/L人生长激素能促进人牙周膜干细胞的增殖和成骨分化。BACKGROUND:Growth hormone has been proven to promote the proliferation and bone differentiation of non-dental mesenchymal stem cells,but the biological effect on human periodontal ligament stem cells is still unclear.OBJECTIVE:To explore the effect of human growth hormone on the proliferation and osteogenic differentiation of human periodontal ligament stem cells.METHODS:Theα-MEM complete medium containing 0(control group),10,100,200μg/L human growth hormone was added to the third generation of human periodontal ligament stem cells.CCK-8 assay was used to detect cell proliferation at 1,3,5,and 7 days.The osteoinductive fluids containing 0(control group),10,100,200μg/L human growth hormone was added to the third generation of human periodontal ligament stem cells.At 7 days,real-time PCR and western blot assay were used to detect the gene and protein expression of the bone differentiation-related factors opn and runx2.At 14 days,Alizarin Red staining and Alizarin Red semi-quantitative detection were used to analyze the mineralization of human periodontal ligament stem cells in the late stage of osteogenic formation.RESULTS AND CONCLUSION:The cell proliferation of 100 and 200μg/L groups was significantly better than that of the control group(P<0.05).The gene and protein expression of opn and runx2 was higher in the 100 and 200μg/L groups than that in the control group(P<0.05).The mineralization of human periodontal ligament stem cells in the 100 and 200μg/L groups was better than that of the control group(P<0.05).Results confirm that in vitro experiments,human growth hormone at concentrations of 100 and 200μg/L can promote the proliferation and osteogenic differentiation of human periodontal ligament stem cells.

关 键 词：干细胞 人牙周膜干细胞 人生长激素 增殖 分化 成骨 因子 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]