化学萃取同种异体肌腱联合同种异体软骨细胞重建兔肩关节前盂唇实验研究  

Experimental study on reconstruction of anterior labrum of shoulder joint by chemical extraction of allogeneic tendon and allogeneic chondrocytes

作　　者：陈国飞 崔磊 陈鹏[2] 李伟[2] 尤田[2] 王琛 江长青[2] 刘岗 CHEN Guofei;CUI Lei;CHEN Peng;LI Wei;YOU Tian;WANG Chen;JIANG Changqing;LIU Gang(Department of Traumatic Osteoarthritis,Shenzhen Hospital,University of Chinese Academy of Sciences,Shenzhen Guangdong,518000,P.R.China;Department of Sports Medicine,Shenzhen Hospital,Peking University,Shenzhen Guangdong,518036,P.R.China)

机构地区：[1]中国科学院大学深圳医院(光明)创伤骨关节科,广东深圳518000 [2]北京大学深圳医院运动医学科,广东深圳518036

出　　处：《中国修复重建外科杂志》2020年第9期1184-1189,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：深圳市三名工程(SZSM201612078);深圳市科技研发资金(JCYJ20180228175214999)。

摘　　要：目的研究化学萃取同种异体肌腱移植联合注射同种异体软骨细胞重建兔肩关节前盂唇损伤的效果。方法成年新西兰大白兔45只,体质量2.5~3.0 kg。取其中15只兔的跟腱,采用化学萃取法进行抗原灭活制备同种异体肌腱,将萃取后肌腱与未处理肌腱行HE染色和Masson染色对比;取膝关节软骨,采用胰蛋白酶法分离培养软骨细胞并行Ⅱ型胶原免疫组织化学染色鉴定。取剩余30只兔制备肩关节前盂唇缺损模型,将同种异体肌腱移植至受损盂唇后,随机分为两组(每组15只),A组移植术后即刻关节内注射同种异体软骨细胞,B组不作任何处理。术后4、6、8周每组各取5只兔的移植肌腱,大体观察后采用HE染色观察细胞核数量,Masson染色观察肌纤维组织胶原纤维的表达情况,AB染色检测移植后糖胺聚糖含量,评估两组同种异体肌腱组织内细胞生长情况。结果 HE染色、Masson染色显示,采用化学萃取法制备的同种异体肌腱抗原被灭活且纤维组织结构保持完好;Ⅱ型胶原免疫组织化学染色示,培养细胞为软骨细胞。肌腱移植术后AB染色示,各时间点A组糖胺聚糖含量均显著高于B组,差异有统计学意义(P<0.05)。术后6周HE染色示,A组肌腱组织内的细胞核数量明显多于B组(t=20.043,P=0.000)。术后6周Masson染色示,A组肌腱组织中的细胞核明显增多,肌纤维及胶原纤维相互交错,组织结构更加紧凑,肌腱组织以蓝染为主;而B组细胞核较少,主要为原移植物的胶原纤维。结论经化学萃取法灭活抗原的同种异体肌腱,能够修复重建兔肩关节前盂唇缺损,且联合同种异体软骨细胞移植能够促进移植肌腱的愈合。Objective To study the effect of chemical extraction of allogeneic tendon and allogeneic chondrocytes for reconstruction of anterior labrum of shoulder joint in rabbits.Methods The body weight of 45 adult New Zealand white rabbits ranged from 2.5 to 3.0 kg.The Achilles tendons of 15 rabbits were taken and the allogeneic tendons were prepared by chemical extraction with antigen inactivation.The extracted tendons were compared with untreated tendons by HE and Masson stainings.Chondrocytes were isolated and cultured by trypsin method and identified by immunohistochemical staining of collagen typeⅡ.The remaining 30 rabbits were used to prepare the model of anterior labrum defect of shoulder joint.After the allogeneic tendon was transplanted to the damaged labrum,the rabbits was randomly divided into two groups(15 in each group).In group A,the allogeneic chondrocytes were injected into the joint immediately after transplantation,while in group B,no treatment was made.At 4,6,and 8 weeks after operation,5 transplanted tendons of each group were taken.After general observation,HE staining was used to observe the number of nuclei,Masson staining was used to observe the expression of collagen fibers in muscle fiber tissues,and AB staining was used to detect the glycosaminoglycan level after transplantation,to evaluate the cell growth in the tissues of the two groups of allogeneic tendon.Results By HE and Masson stainings,the allogeneic tendon antigen prepared by chemical extraction method was inactivated and the fibrous tissue structure was intact;collagen typeⅡimmunohistochemistry staining showed that the cultured cells were chondrocytes.After tendon transplantation,the content of glycosaminoglycan in group A was significantly higher than that in group B(P<0.05).At 6 weeks after operation,HE staining showed that the nuclear in tendon tissue of group A was significantly more than that of group B(t=20.043,P=0.000).Masson staining showed that the number of nuclei in tendon tissue of group A was significantly increased,th

关 键 词：化学萃取法 同种异体肌腱 软骨细胞 盂唇重建 组织工程 兔 

分 类 号：R684[医药卫生—骨科学]