过表达ephrinB2调控犬牙周膜干细胞的成骨能力  

Overexpression of ephrinB2 in canine periodontal ligament stem cells regulates osteogenic differentiation

作　　者：刘雨苗 杨金鑫 朱绍跃 马毓临 夏寒萍 周猛 杨姝蕾 黄泰宁 丁珂欣 刘宗响 Liu Yumiao;Yang Jinxin;Zhu Shaoyue;Ma Yulin;Xia Hanping;Zhou Meng;Yang Shulei;Huang Taining;Ding Kexin;Liu Zongxiang(Xuzhou Medical University,Xuzhou 221004;Dept of Medical,The Affiliated Stomatological Hospital of Xuzhou Medical University,Xuzhou 221002;Dept of Science and Education,The Affiliated Stomatological Hospital of Xuzhou Medical University,Xuzhou 221002)

机构地区：[1]徐州医科大学,徐州221004 [2]徐州医科大学附属口腔医院医务科,徐州221002 [3]徐州医科大学附属口腔医院科教科,徐州221002

出　　处：《安徽医科大学学报》2023年第3期423-428,共6页Acta Universitatis Medicinalis Anhui

基　　金：江苏省自然科学基金青年项目(BK20210080);江苏省大学生创新创业训练计划(202010313003Z)。

摘　　要：目的观察促红细胞生成素肝细胞激酶受体B4/促红细胞生成素肝细胞激酶受体配体B2(EphB4/ephrinB2)信号通路中过表达ephrinB2在比格犬来源的牙周膜干细胞(cPDLSCs)中成骨分化的能力。方法采用Beagle犬前磨牙及磨牙中分离PDLSCs,用EfnB2-GFP-Bsd载体和空载体(GFP-Bsd)转染细胞后进行成骨诱导分化,Western blot检测转染后ephrinB2蛋白表达水平,行CCK-8实验、茜素红S染色实验、碱性磷酸酶染色实验(ALP)、反转录聚合酶链反应(RT-PCR)分别检测EfnB2-cPDLSCs组、空载体对照组(Vector-cPDLSCs)的成骨分化效果。结果CCK-8显示:EfnB2-cPDLSCs组和Vector-cPDLSCs组在细胞增殖上没有差异。茜素红染色实验和ALP染色实验证实:EfnB2-cPDLSCs组较Vector-cPDLSCs显示更高的ALP活性和矿化能力。RT-PCR显示:EfnB2-cPDLSCs组成牙本质/成骨标志物的表达高于Vector-cPDLSCs组。结论通过过表达犬牙周膜细胞中的ephrinB2可提高其成骨分化的能力。Objective Investigate osteogenic differentiation of canine periodontal ligament stem cells(cPDLSCs)via over-expression ephrinB2 in cPDLSCs.Methods cPDLSCs were isolated from the premolars and molars of Beagle.After transfected with EfnB2-GFP-Bsd and GFP-Bsd empty Vector,cPDLSCs were induced to osteogenic differentiation.Western blot was used to invest the expression of ephrinB2 protein.The effect of osteogenic differentiation of EfnB2-cPDLSCs and Vector-cPDLSCs were analyzed by RT-PCR,CCK-8,Alizarin-red S staining and ALP.Results There was no significant difference in cell proliferation between EfnB2-cPDLSCs and Vector-cPDLSCs.While EfnB2-cPDLSCs displayed an enhanced ALP activity and more prominent mineralized nodules compared with Vector-cPDLSCs.The odonto-/osteogenic genes in EfnB2-cPDLSCs were also highly enhanced.Conclusion The results of our study indicated that ephrinB2 gene-transfected cPDLSCs showed enhanced osteogenic differentiation.

关 键 词：犬牙周膜细胞 促红细胞生成素肝细胞激酶受体配体B2 基因转染 成骨分化 

分 类 号：R781.42[医药卫生—口腔医学]