血小板衍生生长因子BB促进人牙周膜干细胞的增殖和成骨分化  

Platelet-derived growth factor BB promotes proliferation and osteogenic differentiation of human periodontal ligament stem cells

作　　者：陈佳娜 聂敏海 胡馨月 刘旭倩 Chen Jiana;Nie Minhai;Hu Xinyue;Liu Xuqian(Department of Periodontics&Oral Mucosal Diseases,Affiliated Stomatology Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Luzhou Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,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牙周黏膜病科,四川省泸州市646000 [2]西南医科大学口颌面修复重建和再生泸州市重点实验室,四川省泸州市646000

出　　处：《中国组织工程研究》2023年第10期1534-1540,共7页Chinese Journal of Tissue Engineering Research

基　　金：四川省干部保健普及应用项目(川干研2022-2101),项目负责人:刘旭倩;四川省泸州市人民政府-西南医科大学科技战略合作重点项目(2019LZXNYDZ10),项目负责人:刘旭倩;西南医科大学校级科研应用基础重点项目(2021ZKZD010),项目负责人:刘旭倩。

摘　　要：背景:人牙周膜干细胞具有多向分化的潜能,在一定条件下能够发生成骨向分化,诱导骨组织再生,因此,人牙周膜干细胞作为骨组织工程种子细胞之一,在牙槽骨缺损骨再生修复中具有良好的研究前景和临床应用价值。目的:探讨血小板衍生生长因子BB在促进人牙周膜干细胞增殖和成骨分化中的功能特征,旨在为牙槽骨缺损的骨再生修复提供相关的实验室依据。方法:人牙周膜干细胞取自就诊于西南医科大学附属口腔医院口腔颌面外科正畸拔牙患者的牙周膜组织,CCK-8法绘制第1-6代人牙周膜干细胞生长曲线,以及不同质量浓度血小板衍生生长因子BB作用下人牙周膜干细胞的增殖曲线。将第3代人牙周膜干细胞分为空白对照组、成骨诱导液组、成骨诱导液+血小板衍生生长因子BB组,分别诱导7,14,21 d,矿化实验观察人牙周膜干细胞钙化结节形成情况,RT-qPCR、Western blot检测Runx2、骨形态发生蛋白2的mRNA和蛋白的相对表达量。结果与结论:(1)CCK-8实验结果显示第2,3代人牙周膜干细胞的对数生长期比其余代次细胞增殖水平显著升高(P <0.05);(2)同一时间点上,100μg/L血小板衍生生长因子BB组的人牙周膜干细胞增殖能力明显优于0,50,200μg/L血小板衍生生长因子BB组(P <0.05);(3)100μg/L血小板衍生生长因子BB与成骨诱导液联合使用对人牙周膜干细胞具有明显的成骨诱导作用,在成骨诱导第14天时,与成骨诱导液组和空白对照组相比,成骨诱导液+血小板衍生生长因子BB组Runx2、骨形态发生蛋白2的mRNA和蛋白相对表达量明显升高(P <0.05);(4)结果表明,血小板衍生生长因子BB能够促进人牙周膜干细胞的增殖及成骨方向分化。BACKGROUND:Human periodontal ligament stem cells have the potential for multidirectional differentiation.Under certain conditions,they can differentiate towards osteogenesis and induce bone tissue regeneration.Therefore,human periodontal ligament stem cells,as one of bone tissue engineering seed cells,have good research prospects and clinical application value in bone regeneration and repair of alveolar bone defects caused by periodontal disease.OBJECTIVE:To explore the functional characteristics of platelet derived growth factor-BB in promoting the proliferation and osteogenic differentiation of human periodontal ligament stem cells,aiming to provide relevant laboratory evidence for bone regeneration and repair of alveolar bone defects.METHODS:Human periodontal ligament stem cells were taken from the periodontal ligament tissue of orthodontic extraction patients in the Department of Oral and Maxillofacial Surgery,Affiliated Stomatology Hospital of Southwest Medical University.The growth curve of passages 1-6 human periodontal ligament stem cells and the proliferation curve of human periodontal ligament stem cells under the action of different concentrations of platelet-derived growth factor BB were drawn by CCK-8 assay.Passage 3 human periodontal ligament stem cells were divided into blank control group,osteoinduction liquid group,osteoinduction liquid + platelet-derived growth factor BB group.The formation of mineralized nodules after induction for 7,14,and 21 days was observed by mineralized experiment.RT-qPCR and western blot assay were used to detect the relative expression of Runx2 and bone morphogenetic protein 2 mRNA and protein.RESULTS AND CONCLUSION:(1) CCK-8 assay results showed that the passages 2 and 3 human periodontal ligament stem cells in logarithmic growth phase were significantly higher than that of the other passages(P < 0.05).(2) At the same time point,the proliferation of human periodontal ligament stem cells in 100 μg/L platelet-derived growth factor BB group was significantly better than

关 键 词：人牙周膜干细胞 血小板衍生生长因子 增殖 成骨 牙槽骨缺损 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R781.4