BMP-2、TGF-β1、干扰素γ参与人牙髓干细胞增殖及成骨向分化的机制探究  

Mechanism of BMP-2,TGF-β1 and IFN-γin proliferation and osteogenic differentiation of human dental pulp stem cells

作　　者：邵贝贝 段铸辉 贺龙龙[2] SHAO beibei;DUAN Zhuhui;HE Longlong(Department of Stomatology,Luoyang Central Hospital Affiliated to Zhengzhou University,Henan,Luoyang,471000 China;Department of dental implant,School of Stomatology,Xi'an Jiaotong University,Shaanxi Clinical Research Center for dental and maxillofacial diseases,Shaanxi Key Laboratory of craniomaxillofacial precision medicine,Shanxi,Xi'an,710000 China)

机构地区：[1]郑州大学附属洛阳中心医院口腔科,河南洛阳471000 [2]西安交通大学口腔医学院口腔种植科陕西省牙颌面疾病临床研究中心陕西省颅颌面精准医学研究重点实验室,陕西西安710000

出　　处：《实验与检验医学》2022年第4期391-395,共5页Experimental and Laboratory Medicine

基　　金：陕西省重点研发计划,编号2018SF-003。

摘　　要：目的 探究人骨形态发生蛋白-2(BMP-2)、转化生长因子-β1(TGF-β1)、干扰素γ(IFN-γ)参与人牙髓干细胞增殖及成骨向分化的机制。方法 选取因阻生或正畸减数拔除的健康恒牙,经组织块酶消化法分离并提取牙髓干细胞进行原代培养,光镜观察形态学变化,取对数生长期人牙髓干细胞,加入不同浓度BMP-2(50 ng/mL、100 ng/mL、150 ng/mL)、TGF-β1(1 ng/mL、5 ng/mL、10 ng/mL)、IFN-γ(50 U/mL、100 U/mL、200 U/mL),同时设置阴性对照组,分别在第1 d、3 d、7 d检测细胞增殖情况(OD值),第7 d、14 d检测成骨向分化指标[碱性磷酸酶(ALP)、Ⅰ型胶原蛋白(COL-Ⅰ)]表达情况,Spearman分析BMP-2、TGF-β1、IFN-γ与OD值、成骨向分化指标相关性。结果 (1)当BMP-2、TGF-β1、IFN-γ浓度分别为100 ng/mL、5 ng/mL、200 U/mL时,OD值达到峰值;(2)第7 d联合组OD值高于对照组(P<0.05);(3)第7 d、14 d BMP-2组、TGF-β1组、IFN-γ组、联合组ALP、COL-Ⅰ水平高于对照组(P<0.05);⑷Spearman分析得知,BMP-2、TGF-β1、IFN-γ与OD值、ALP、COL-Ⅰ呈正相关(P<0.05)。结论 BMP-2、TGF-β1、IFN-γ联合能显著促进人牙髓干细胞增殖及成骨向分化,对牙髓及牙周膜等纤维结缔组织的再生具有一定理论意义。Objective To investigate the mechanisms of human bone morphogenetic protein-2(BMP-2),transforming growth factor-β1(TGF-β1),and interferonγ(IFN-γ)in the proliferation and differentiation of human dental pulp stem cells.Methods Healthy permanent teeth extracted by impaction or orthodontic reduction were selected,and dental pulp stem cells were isolated and extracted by tissue block enzyme digestion for primary culture,the morphological changes were observed under a light microscope,and human dental pulp stem cells in logarithmic growth phase were taken,added different concentrations of BMP-2(50 ng/mL,100 ng/mL,150 ng/mL),TGF-β1(1 ng/mL,5 ng/mL,10 ng/mL),IFN-γ(50 U/mL,100 U/mL,200 U/mL).At the same time,a negative control group was set up,and the cell proliferation(OD value)was measured on the 1st,3rd,and 7th days,the expression of osteogenic differentiation indicators[alkaline phosphatase(ALP),typeⅠcollagen(COL-Ⅰ)]was detected on the 7th and 14th days,respectively,Spearman analyzed the correlations between BMP-2,TGF-β1,IFN-γand OD values and osteogenic differentiation indicators.Results⑴When the concentrations of BMP-2,TGF-β1,and IFN-γwere 100 ng/mL,5 ng/mL,and 200 U/mL,the OD value reached its peak;⑵The OD value of the combined group on the 7th day was higher than that of the control group(P<0.05);⑶The levels of ALP and COL-Ⅰof the BMP-2 group、TGF-β1 group、IFN-γgroup and combined group on the 7th and 14th days were higher than the control group(P<0.05);⑷Spearman analysis showed that BMP-2,TGF-β1,and IFN-γwere positively correlated with OD values,ALP,and COL-I(P<0.05).Conclusion The combination of BMP-2,TGF-β1 and IFN-γcan significantly promote human dental pulp stem cell proliferation and osteogenic differentiation,and has certain theoretical significance for the regeneration of fibrous connective tissue such as dental pulp and periodontal ligament.

关 键 词：牙髓干细胞 增殖 成骨向分化 BMP-2 TGF-Β1 

分 类 号：R394.2[医药卫生—医学遗传学]