骨髓间充质干细胞荷载微小RNA-29对大鼠胫骨骨缺损的治疗作用  

Effect of microRNA-29 loading on bone marrow mesenchymal stem cells on tibial bone defect in rats

作　　者：石宁宁[1] 谢艳[2] 单海民[1] 王新江[1] 肖建伟 郑吉全 Shi Ningning;Xie Yan;Shan Haimin;Wang Xinjiang;Xiao Jianwei;Zheng Jiquan(Department of Hand External Microscopic Orthopedics,Luoyang Orthopedic-Traumatological Hospital of Henan Province,Henan Provincial Orthopedic Hospital,Luoyang 471002,China;Luoyang Orthopedic-Traumatological Hospital Of Henan Province,Henan Provincial Orthopedic Hospital Biomedical Engineering Research Office,Luoyang 471002,China)

机构地区：[1]河南省洛阳正骨医院、河南省骨科医院手外显微骨科,洛阳471002 [2]河南省洛阳正骨医院、河南省骨科医院生物医学工程研究室,洛阳471002

出　　处：《中华实验外科杂志》2024年第10期2301-2304,共4页Chinese Journal of Experimental Surgery

基　　金：河南省科技发展项目(212102310855)。

摘　　要：目的:探讨骨髓间充质干细胞荷载微小RNA(miR)-29对大鼠胫骨骨缺损的治疗作用。方法:将购自河南实验动物中心的30只SD大鼠按照随机数字表法随机分为对照组、模型组和治疗组,每组10只,模型组和治疗组大鼠采用手术制造胫骨骨缺损模型,对照组大鼠仅手术,不做骨缺损手术。治疗组大鼠骨缺损部位注射荷载miR-29的骨髓间充质干细胞,对照组和模型组只注射骨髓间充质干细胞。治疗8周后,X线分析骨缺损部位愈合情况;苏木精-伊红(HE)染色和Masson染色测定新生骨面积百分比;采用抗酒石酸酸性磷酸酶染色观察破骨细胞分化程度;采用酶联免疫吸附实验分析3组大鼠降钙素、Ⅰ型胶原C-末端肽、血管内皮生长因子等水平;Micro-CT检测骨缺损区内骨体积分数(BV/TV),组织骨密度(tBMD)和骨小梁数量(Tb.N)等参数的变化。组间计量数据比较采用单因素方差分析。结果:治疗组大鼠X线片评分(3.87±0.75)高于模型组(2.35±0.57),差异统计学意义(t=3.012,P<0.05)。治疗组大鼠胶原纤维面积比和新生骨面积比[(12.20±2.78)%、(35.87±3.59)%]高于模型组[(5.70±1.16)%、(20.68±2.14)%],差异有统计学意义(t=6.822、11.480,P<0.05)。对照组大鼠骨缺损部位破骨细胞数量[(6.10±1.52)个]低于模型组[(12.50±2.22)个],差异有统计学意义(t=7.508,P<0.05)。治疗组大鼠骨缺损部位破骨细胞数量[(8.20±1.23)个]低于模型组[(12.50±2.22)个],差异有统计学意义(t=5.352,P<0.05)。治疗组大鼠骨缺损部位降钙素和Ⅰ型胶原C-末端肽水平[(8.62±0.70)、(21.00±4.13)ng/ml]低于模型组[(16.88±2.69)、(29.78±4.46)ng/ml],差异有统计学意义(t=9.397、4.566,P<0.05)。治疗组大鼠血管内皮生长因子水平[(74.72±6.93)ng/ml]高于模型组[(55.49±8.12)ng/ml],差异有统计学意义(t=5.695,P<0.05)。治疗组大鼠骨缺损部位tBMD和BV/TV和Tb.N[(0.26±0.04)g/cm^(2)、(22.20±2.04)%、(1.52±0.15)/mm]高于模型组[(0.14±0.02Objective:To investigate the therapeutic effect of microRNA(miR)-29 loaded bone marrow mesenchymal stem cells on tibial bone defect in rats.Methods:Thirty SD rats from Henan experimental animal center were randomly divided into control group,model group and treatment group according to random number table method,with 10 rats in each group.The model group and treatment group were operated to create the tibial bone defect model,while the control group was operated only without bone defect operation.Bone marrow mesenchymal stem cells loaded with miR-29 were injected into the bone defect site of rats in the treatment group,while only bone marrow mesenchymal stem cells were injected into the control group and model group.After 8 weeks of treatment,the healing of bone defect was analyzed by X-ray.The percentage of new bone area was determined by hematoxylin and eosin(HE)staining and Masson staining.The differentiation degree of osteoclasts was observed by tartrate-resistant acid phosphatase staining.The levels of calcitonin,C-terminal peptide of typeⅠcollagen and vascular endothelial growth factor were analyzed by enzyme-linked immunosorbent assay.The changes of bone volume fraction(BV/TV),tissue bone mineral density(tBMD)and trabecular number(Tb.N)were measured by Micro-CT.One-way analysis of variance was used to compare the measurement data between groups.Results:The X-ray score of rats in the treatment group(3.87±0.75)was significantly higher than that of rats in the model group(2.35±0.57,t=3.012,P<0.05).The area ratio of collagen fibers and the area ratio of new bone in the treatment group[(12.20±2.78)%,(35.87±3.59)%]were significantly lower than those in the model group[(5.70±1.16)%,(20.68±2.14)%,t=6.822,11.480,P<0.05].The number of osteoclasts in the bone defect site in the control group[(6.10±1.52)cells]was significantly lower than that in the model group[(12.50±2.22)cells,t=7.508,P<0.05].The number of osteoclasts in the treatment group was the number of osteoclasts in the bone defect site of rats[(8.2

关 键 词：骨髓间充质干细胞 微小RNA-29 胫骨骨缺损 骨愈合 

分 类 号：R681.8[医药卫生—骨科学]