骨形态发生蛋白-2提高骨髓间充质干细胞对骨质疏松牙槽骨缺损的修复作用  

Bone morphogenetic protein-2 enhances the repairing effect of bone marrow mesenchymal stem cells on osteoporotic alveolar bone defect

作　　者：吴萌萌[1] 吕晓丹[1] 张扬[2] 戴东晓[1] 王会超[3] WU Meng-meng;LV Xiao-dan;ZHANG Yang;DAI Dong-xiao;WANG Hui-chao(Department of Stomatology,Shijiazhuang Second Hospital,Shijiazhuang 050000,China;Department of Medical Imaging,Hebei Medical Univer-sity Stomatology Hospital,Shijiazhuang 050000,Chin)

机构地区：[1]石家庄市第二医院口腔科,石家庄050000 [2]河北医科大学口腔医院医学影像科,石家庄050000 [3]石家庄市第二医院口腔科国际部,石家庄050000

出　　处：《口腔颌面修复学杂志》2025年第2期81-89,共9页Chinese Journal of Prosthodontics

基　　金：河北省医学科学研究课题计划(项目编号:20201339)。

摘　　要：目的:探究骨形态发生蛋白-2(bone morphogenetic protein 2,BMP-2)诱导骨髓间充质干细胞对骨质疏松牙槽骨缺损的修复作用。方法:分离小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),流式细胞仪鉴定表面标志物;BMP-2腺病毒转染至BMSCs,分为NC组、Ad-EGFP组和AdBMP-2/EGFP组,荧光显微镜下观察转染效率,逆转录-定量聚合酶链反应(reverse transcription-quantitative polymerase chain reaction,RT-qPCR)检测细胞中BMP-2 mRNA相对表达量;茜素红染液检测BMSCs成骨能力;制备骨质疏松大鼠模型并用Micro-CT扫描;制备牙槽骨缺损大鼠模型,随机分为Sham组(正常大鼠双侧骨缺损内植入明胶海绵)、OP组(OP大鼠双侧骨缺损内植入明胶海绵)、BMSCs+Ad-EGFP组(OP大鼠双侧骨缺损内植入BMSCs+Ad-EGFP明胶海绵复合体)和BMSCs+Ad-BMP-2/EGFP组(OP大鼠双侧骨缺损内植入BMSCs+Ad-BMP-2/EGFP明胶海绵复合体)。采用微型计算机断层扫描(micro-computed tomography,micro-CT)检测各组大鼠骨缺损情况;血红素-伊奥红素染色(hematoxylin-eosin,HE)检测牙槽骨组织病理学变化;蛋白质印迹法检测牙槽骨组织中碱性磷酸酶(alkaline phosphatase,ALP)、骨钙素(osteocalcin,OCN)、骨形成蛋白2(runt-related transcription factor 2,Runx2)蛋白表达。结果:流式细胞仪鉴定BMSCs表面CD29、CD90、CD105、CD44抗体呈阳性,CD45、CD34、CD19抗体呈阴性,表明分离提取的细胞即为BMSCs。和NC组相比,Ad-EGFP组和Ad-BMP-2/EGFP组转染效率均超过90%,提示AdEGFP和Ad-BMP-2/EGFP转染到BMSCs中。Ad-BMP-2/EGFP组细胞中BMP-2 mRNA表达明显高于NC组(P<0.05)。茜素红染色显示,Ad-EGFP组仅有少量的钙化结节,Ad-BMP-2/EGFP组可见大量的矿化结节,且被染成红色。造模组骨密度、Tb.Th、Tb.N、BV/TV均低于Sham组,Tb.Sp高于Sham组(P<0.05),提示OP模型制备成功。OP组BMD、BV/TV、组织病理学评分、牙槽骨组织中ALP、OCN、Runx2蛋白表达明显低于Sham组,Tb.Sp明显高�Objective:Investigation of the reparative effect of bone marrow mesenchymal stem cells induced by bone morphogenetic protein 2(BMP-2)on alveolar bone defects in osteoporotic periodontitis.Methods:Mice bone marrow mesenchymal stem cells(BMSCs)were isolated and the surface markers were identified using flow cytometry.BMSCs were then transfected with BMP-2 adenovirus,and divided into NC group,Ad-EGFP group,and Ad-BMP-2/EGFP group.Transfection efficiency was observed under fluorescence microscope,and the relative mRNA expression of BMP-2 in cells was detected by Reverse Transcription-quantitative Polymerase Chain Reaction(RT-qPCR).Von Kossa staining was used to evaluate the osteogenic ability of BMSCs.Osteoporotic rat model was established and scanned using Micro-CT to compare trabecular bone thickness(Tb.Th),trabecular bone number(Tb.N),bone volume/total volume(BV/TV),and trabecular separation(Tb.Sp)between the two groups.Periodontal bone defect rat model was created,and rats were randomly divided into Sham group(gelatin sponge implanted in bilateral bone defects of normal rats),OP group(gelatin sponge implanted in bilateral bone defects of osteoporotic rats),BMSCs+Ad-EGFP group(gelatin sponge composite of BMSCs+Ad-EGFP implanted in bilateral bone defects of osteoporotic rats),and BMSCs+Ad-BMP-2/EGFP group(gelatin sponge composite of BMSCs+Ad-BMP-2/EGFP implanted in bilateral bone defects of osteoporotic rats).Micro-CT was used to evaluate the bone defect condition of each group.Hematoxylin-Eosin(HE)staining was performed to examine the histopathological changes in periodontal bone tissue.Protein expression of alkaline phosphatase(ALP),osteocalcin(OCN),and runt-related transcription factor 2(Runx2)in periodontal bone tissue was detected using Western blotting.Results:Flow cytometry analysis revealed that the surface markers CD29 and CD90 were positive,while CD45 and CD34 were negative,indicating that the isolated cells are bone marrow mesenchymal stem cells(BMSCs).Compared with the NC group,the transfection efficien

关 键 词：骨形态发生蛋白-2 骨髓间充质干细胞 成骨 骨质疏松牙槽骨缺损 

分 类 号：R780.2[医药卫生—口腔医学]