antagomir-30d转染BMSCs联合CPC支架材料的异位成骨作用  

Ectopic study of calcium phosphate cement seeded with antagomir-30d transfected BMSCs

作　　者：陈威[1] 汪艳 金琼[2] 黄慧[3] CHEN Wei;WANG Yan;JIN Qiong;HUANG Hui(Department of Pediatric Dentistry,School&Hospital of Stomatology Wenzhou Medical University,Wenzhou 325000,China;Department of Implant Prosthodontics,School&Hospital of Stomatology Wenzhou Medical University,Wenzhou 325000,China;Department of Prosthodontics,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]温州医科大学附属口腔医院儿童口腔科,浙江温州325000 [2]温州医科大学附属口腔医院种植科,浙江温州325000 [3]上海交通大学附属第九人民医院修复科,上海200011

出　　处：《温州医科大学学报》2023年第1期1-6,共6页Journal of Wenzhou Medical University

基　　金：国家自然科学基金项目(81170988)。

摘　　要：目的:构建antagomir-30d/种子细胞骨髓间充质干细胞(BMSCs)/磷酸钙骨水泥(CPC)复合物,并将该复合物植入裸鼠皮下,研究antagomir-30d促进体内骨形成的作用。方法:将转染有150 nmol/L antagomir-30d、NC的BMSCs以及未经转染的空白BMSCs转移到CPC支架进行孵育,后植入到BALB/c-nu裸鼠皮下,以此研究裸鼠异位成骨。术后2、4、8周分别取出植入体。2周时,取出植入体,提取RNA进行RT-PCR检测,分析成骨基因碱性磷酸酶(ALP)、骨钙素(OC)以及Runt相关转录因子2(RUNX2)mRNA表达情况。此外,4周和8周的植入体分别进行苦味酸品红组织学染色和组织形态学分析新骨形成的情况。结果:植入2周RT-PCR结果显示,各成骨基因mRNA水平的表达量中转染有antagomir-30d组最高,并显著高于另外2组(P<0.05)。组织学染色结果表明,植入后4周,antagomir-30d组有少量新骨形成,而另外2组则少有新骨形成。组织形态学分析显示,新骨面积百分比antagomir-30d组为1.28%±0.19%。植入后8周,3组均可见明显新骨形成,但antagomir-30d组新骨形成量明显大于NC组及空白对照组(P<0.05),各组新骨面积百分比分别为17.79%±1.15%,1.82%±0.53%,2.33%±0.52%。结论:antagomir-30d可诱导裸鼠体内异位成骨,antagomir-30d/BMSCs/CPC复合物有希望作为组织工程复合物用于颌骨骨缺损及其他类型骨缺损的修复。Objective:To constructed a compound of antagomir-30d/bone marrow mesenchymal stem cells(BMSCs)/calcium phosphate cement(CPC)and implant s.c.in BALB/c-nu mice to demonstrate that antagomir-30d could promote ectopic osteogenesis.Methods:The BMSCs transfected with the optimum concentration of antagomir-30d/NC/blank was loaded on calcium phosphate cement scaffolds and s.c.was implanted into BALB/c-nu mice.Implants were removed after 2,4,8 week(wk)respectively.2-wk implants were subjected to RNA extraction,and then gene expression of osteoblast marker genes alkaline phosphatase(ALP),osteocalcin(OC)and Runt-related transcription factor 2(RUNX2)was analyzed.4-wk and 8-wk implants were subjected to histological staining and histomorphometric analysis was made to observe new bone formation.Results:Gene expression of osteoblast marker genes was analyzed after 2 wk implantation.RT-PCR analysis revealed upregulation of ALP,OC and RUNX2 in the antagomir-30d-treated implants compared with implants transfected with miR negative control and the un-transfected group.The histological staining revealed that the antagomir-30d-treated implants had a small amount of new bone formation at 4 wk.Implants transfected with miR negative control and the un-transfected group showed new bone formation barely.Histomorphometric analysis showed that the percentage of new bone area of antagomir-30d-treated implants was 1.28%±0.19%.At 8 wk,new bone formation was clearly visible in all groups,while the new bone formation of antagomir-30d-treated group was more significant than NC and un-transfected groups(P<0.05).New bone area percentage was 17.79%±1.15%,1.82%±0.53%,2.33%±0.52%respectively.Conclusion:Antagomir-30d accelerates ectopic bone formation in vivo,and a compound of antagomir-30d/BMSCs/CPC is promising as tissue engineering complexes in repairing jaw bone defects.

关 键 词：miR-30d拮抗剂 骨髓基质干细胞 磷酸钙骨水泥 异位成骨 

分 类 号：R780.1[医药卫生—口腔医学]