牙源性间充质干细胞对成骨前体细胞成骨分化的影响  

Effects of Odontogenic Mesenchymal Stem Cells on Osteogenic Differentiation of Osteoblast Precursor Cells

作　　者：李慧 马博 余日月 郑明珠 宋慧妍 LI Hui;MA Bo;YU Ri-yue;ZHENG Ming-zhu;SONG Hui-yan(Department of Stomatology,Beijing Shijitan Hospital,Capital Medical University,Beijing,100038,China)

机构地区：[1]首都医科大学附属北京世纪坛医院口腔科,北京100038

出　　处：《现代生物医学进展》2021年第2期205-209,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年基金项目(81901001)。

摘　　要：目的:探讨牙源性间充质干细胞对成骨前体细胞成骨分化的影响。方法:将小鼠成骨前体细胞MC3T3-E1分为两组,观察组为牙源性间充质干细胞与MC3T3-E1细胞共培养,对照组为单一MC3T3-E1细胞培养。采用CCK-8法检测细胞增殖水平,采用酶联免疫法检测碱性磷酸酶(Alkaline phosphatase,ALP)活性并进行茜素红染色,采用qRT-PCR、Western blot检测ALP与骨桥素(osteopontin,OPN) m RNA与蛋白表达水平。结果:细胞共培养1 d与3 d后,观察组的细胞增殖指数、ALP活性显著高于对照组(P<0.05)。与对照组相比,观察组的矿化结节显著增加,经茜素红染色呈红褐色。细胞共培养1 d与3 d后,观察组的ALP、OPN m RNA与蛋白相对表达水平显著高于对照组(P<0.05)。结论:牙源性间充质干细胞能促进成骨前体细胞的ALP、OPN表达,提高ALP活性,增加细胞增殖能力,诱发矿化,从而促进成骨分化。Objective: To investigate the effects of odontogenic mesenchymal stem cells on osteogenic differentiation of osteoblast precursor cells. Methods: The MC3 T3-E1 mouse osteoblast precursor cells were divided into two groups. The observation group were co-cultured with odontogenic mesenchymal stem cells and MC3 T3-E1 cells. The control group were cultured with single MC3 T3-E1 cell. CCK-8 method were used to detect cell proliferation level, enzyme-linked immunoassay were used to detect Alkaline phosphatase(ALP) activity and alizarin red staining, and qRT-PCR and Western blot were used to detect ALP and osteopontin(osteopontin, OPN)m RNA and protein expression levels. Results: After the cells were co-cultured for 1 and 3 days, the cell proliferation index and ALP activity in the observation group were significantly higher than those in the control group(P<0.05). Compared with the control group, the mineralized nodules in the observation group increased significantly, and they were reddish brown after stained with alizarin red. After co-culture of cells for 1 and 3 days, the relative expression levels of ALP, OPN m RNA and protein in the observation group were significantly higher than those in the control group(P<0.05). Conclusion: Dental derived mesenchymal stem cells can promote the expression of ALP and OPN of osteoblast precursor cells, increase ALP activity, increase cell proliferation ability, induce mineralization, and thus promote osteogenic differentiation.

关 键 词：牙源性间充质干细胞 成骨前体细胞 碱性磷酸酶 骨桥素 成骨分化 

分 类 号：R-33[医药卫生] R331.2