低强度脉冲超声促进BMSCs定向分化对兔软骨缺损影响研究  

Effect of low-intensity pulsed ultrasound on directional differentiation of BMSCs on cartilage defects in rabbits

作　　者：程光[1] 吕敏[2] 刘爱华[3] CHENG Guang;LYU Min;LIU Ai-hua(Department of Ultrasound,Fourth People’s Hospital of Tai’an,Taian 271000,P.R.China;Tai’an Central Hospital,Taian 271000,P.R.China)

机构地区：[1]泰安市第四人民医院超声科,山东泰安271000 [2]泰安市中心医院超声科,山东泰安271000 [3]泰安市中心医院中心实验室,山东泰安271000

出　　处：《社区医学杂志》2020年第12期858-864,共7页Journal Of Community Medicine

摘　　要：目的膝关节内关节面软骨缺损是临床常见病,其治疗主要集中膝关节镜下滑膜清理联合微骨折处理,但对于缺损较大的患者,效果不佳。本研究探究低强度脉冲超声促进骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)定向分化在兔软骨缺损模型中的修复作用,以期为临床上利用超声技术对骨缺损的治疗提供新思路和方法。方法以健康清洁级新西兰大白兔为研究对象,提取和培养兔源性BMSCs,采用免疫荧光染色和流式细胞仪鉴定BMSCs,构建新西兰大白兔膝关节软骨缺损模型。根据处理方案的不同进行分组,BMSCs组单纯膝关节注射BMSCs细胞,超声组利用低强度脉冲超声进行处理,联合组利用低强度脉冲超声技术联合膝关节注射BMSCs细胞,对照组未做处理。采用组织大体学方法和Micro-CT检测软骨缺损的修复情况。采用HE、番红染色和Ⅱ型胶原(CollagenⅡ)及多聚蛋白多糖(Aggrecan)免疫组织化学染色检测软骨缺损模型。采用RT-qPCR定量检测CollagenⅡ及Aggrecan mRNA表达水平。结果兔膝关节软骨缺损修复组织Ramallal评分结果显示,处理6周后联合组评分为(7.69±0.71)分,高于对照组的(2.89±0.34)分,F=9.389,P=0.016;12周后,联合组评分为(10.22±0.69)分,高于对照组的(4.11±0.46)分,F=9.786,P=0.019。HE和番红染色结果显示,处理6周联合组评分分别为(7.7±0.3)和(4.5±0.9)分,高于对照组的(5.9±0.7)和(1.4±0.2)分,F值分别为9.987和10.981,P值分别为0.021和<0.001;12周后,联合组评分分别为(8.7±1.1)和(5.6±0.6)分,高于对照组的(6.0±0.5)和(1.5±0.2)分,F值分别为14.076和18.008,均P<0.001。免疫组织化学染色结果显示,处理6周联合组CollagenⅡ和Aggrecan相对表达量分别为3.3±0.9和3.6±1.1,高于对照组的1.1±0.1和1.7±0.5,F值分别为15.116和19.987,均P<0.001;12周后,联合组分别为4.7±1.6和5.3±1.2,高于对照组的1.2±0.3和1.8±0.7,F值分别为19.221和16.654,均P<0.001。OBJECTIVE The articular cartilage defect in the knee joint is a common clinical disease.The treatment of this type of disease mainly focuses on knee arthroscopic synovial membrane cleaning combined with microfracture treatment.However,for patients with larger defects,the effect is not good.This study explored the role of low-intensity pulsed ultrasound in the repair of BMSCs directional differentiation in rabbit cartilage defect models,with a view to providing new ideas and methods for the clinical treatment of bone defects using ultrasound.METHODS Taking healthy and clean New Zealand white rabbits as research objects,extract and culture rabbit bone marrow mesenchymal stem cells(BMSCs),identify BMSCs with immunofluorescence staining and flow cytometry,and construct a New Zealand white rabbit knee cartilage defect model.In the BMSCs group,knee joints were injected with BMSCs alone,and the ultrasound group was treated with low-intensity pulsed ultrasound.The combined group was treated with low-intensity pulsed ultrasound and knee joints were injected with BMSCs.Tissue gross method and Micro CT were used to detect the repair of cartilage defects.The cartilage defect model was detected by HE staining,saffron staining,collagenⅡ and collagen aggrecan(Aggrecan)immunohistochemical staining.RT-qPCR was used to quantitatively detect the expression level of collagenⅡ and Aggrecan.RESULTS The Ramallal score of rabbit knee joint cartilage defect repair tissue showed that the score of the combined group after treatment for 6 weeks was(7.69±0.71)points,which was higher than that of the control group(2.89±0.34)points,F=9.389,P=0.016.After treatment for 12 weeks,the score of the combined group was(10.22±0.69)points,which was higher than that of the control group(4.11±0.46)points,F=9.786,P=0.019.The results of HE staining and saffron staining showed that the scores of the combined group at 6 weeks after treatment were(7.7±0.3)and(4.5±0.9)points,which were higher than the control group(5.9±0.7)and(1.4±0.2)points,and the

关 键 词：软骨缺损 低强度脉冲超声 骨髓间充质干细胞 修复 

分 类 号：R684[医药卫生—骨科学]