组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2012, Vol. 8 ›› Issue (2): 65-68.doi: 10.3969/j.issn.1673-0364.2012.02.002

• 论著 • 上一篇    下一篇

P17-BMP2多肽促进大鼠骨髓间充质干细胞成骨分化的实验研究

李佳滨，张文杰，崔福斋，杜晓岩   

  1. 佳木斯大学附属第二医院、口腔医院整形外科；上海市组织工程研究重点实验室；上海交通大学医学院附属第九人民医院整复外科；清华大学材料科学与工程系
  • 发布日期:2020-07-23

Effect of P17-BMP2 Peptide in the Osteogenic Differentiation of BMSCs in Rats

LI Jiabin1,ZHANG Wenjie2,CUI Fuzhai3,DU Xiaoyan1   

  1. 1 Department of Orthopaedics,Hospital of Stomatology,Jiamusi University,Jiamusi 154004,China;2 Shanghai Key Laboratory of Tissue Engineering,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China;3 Department of Materials Science and Engineering,Tsinghua University,Beijing 100084,China.
  • Published:2020-07-23
  • Contact: 佳木斯大学研究生创新科研项目　　（YJSCX2011-026JD）

摘要： 目的探讨P17-BMP2对大鼠骨髓间充质干细胞(BMSCs)增殖和成骨分化的影响。方法体外培养Wistar大鼠BMSCs,分4组培养:对照组(A组)、成骨诱导组(B组)、成骨诱导+P17-BMP2组(C组)、单纯P17-BMP2组(D组)。A组:采用普通DMEM培养基培养;B组:含地塞米松10-7mol/L、Vit C 10 mg/L及β-甘油磷酸钠(β-sodium glycerophos-phate,β-GP)10 mmol/L的DMEM培养基培养;C组:成骨诱导液+P17-BMP2(10μg/mL)培养;D组:含P17-BMP2(10μg/mL)的DMEM培养基培养。CCK-8法检测培养第1、3、5、7天时细胞增殖情况;培养1周后,碱性磷酸酶染色(Alkaline phosphatase,ALP),实时定量PCR检测骨钙素(Osteocalcin,OCN)、骨桥蛋白(Osteopontin,OPN)、转录因子(Runt-related transcription factor 2,Runx2)的mRNA表达。结果与A组相比,D组对大鼠BMSCs增殖无影响,而B组对BMSCs细胞增殖具有促进作用,C组细胞增殖能力显著提高;各组碱性磷酸酶染色均为阳性,但染色强度有差异,C组>B组>D组>A组;Q-PCR检测结果显示:与A相比,B组和C组中所有检测基因的表达均显著提高。结论 P17-BMP2能协同成骨诱导剂促进BMSCs的成骨分化。

关键词: 骨髓间充质干细胞, P17-骨形态发生蛋白2, 增殖, 成骨分化

Abstract: Objective To investigate the effects of P17-BMP2 peptide on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs derived from 4-week-old Wistar rats were isolated and expanded.Four groups were set in this study: Group A,which were cultured in regular medium(DMEM with 10% fetal calf serum);Group B,which were cultured in the regular medium with dexamethasone 107 mol/L,Vitamin C 10 mg/L and beta-glycerophosphate 10 mmol/L(induction medium);Group C,which were cultured in induction medium with P17-BMP2 10 μg/mL;and Group D,which were cultured in regular medium with P17-BMP2 10 μg/mL.Cell proliferation were measured at 1、3、5、7 days after culture by CCK-8 kit.Alkaline phosphatase(AKP) staining were performed 7 days after culture.The expressions of osteocalcin(OCN),osteopontin(OPN),runt-related transcription factor 2(Runx2) were evaluated by Q-PCR.Results Compared with group A,the proliferation of BMSCs were not promoted in group D,but promoted in group B and highly enhanced in group C.The alkaline phosphatase staining were all positive in each group,but the dyeing intensity were different: group C > group B > group D > group A.Compared with group A,the expressions of OCN,OPN and Runx2 were

Key words: Bone marrow mesenchymal stem cells, P17-BMP2 peptide, Proliferation, Osteogenic differentiation

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