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___________用组织工程学方法探讨兔佐剂性关节炎关节软骨修复的实验研究_________

Experimental research by tissue engineering methods for repairing cartilage defects in adjuvant arthritis in rabbits

作者:               张旭东  逯卓卉  陈伟  蒋雯雯  韦登明          
单位:           宁波大学医学院病理学与病原生物学系(浙江宁波315211)    
关键词:           佐剂性关节炎;软骨缺损;组织工程;自体骨髓间充质干细胞;软骨组织学评分      
分类号:           R318.08    
出版年·卷·期(页码):2016·35·2(185-190)
摘要:

目的 探讨海藻酸盐支架超微结构及软骨形态变化与组织工程修复佐剂性关节炎软骨缺损的关系,为临床应用组织工程方法治疗类风湿性关节炎(rheumatoid arthritis, RA)提供一定的实验依据。方法 将24只新西兰兔随机均分为假手术对照组,关节炎模型组和软骨细胞-海藻酸钙支架复合物治疗组。对模型组和治疗组膝关节处注射0.5mL完全弗氏佐剂诱导关节炎,假手术组注射等量生理盐水。抽取治疗组骨髓5mL,Percoll密度梯度分离出自体骨髓间充质干细胞,体外培养纯化,并诱导分化成软骨细胞。与海藻酸钙支架混合培养,对支架进行扫描电镜观察,并将混合物回注相应关节腔内治疗1个月。对各组兔膝关节进行组织学评分检测软骨缺损修复结果。结果 海藻酸盐支架电镜扫描显示支架具有一定的孔隙,有利于营养物质的进入,便于细胞的增殖分化。软骨组织评分结果显示软骨纤维化减轻,关节腔内积液消失,软骨缺损得到一定修复。结论 软骨组织评分显示海藻酸钙复合工程化软骨细胞对兔佐剂性关节炎软骨缺损有一定的修复作用,其机制可能与支架有利于软骨细胞生长发育有关。

Objective To explore ultrastructure of alginate scaffolds, morphological changes of cartilage and the relation with tissue engineering repairing adjuvant arthritis in order to provide certain experimental basis for the clinical application of tissue engineering. Methods We divided the rabbits into three groups: control group (n=8), arthritis model group (n=8), cartilage cells-scaffold complex treatment group (n=8). We injected 0.5mL adjuvant into the rabbits’ knee of model group and treatment group to induce arthritis, and 0.5mL physiological saline into control group. We extracted 5mL of bone marrow of treatment group, and separated the bone marrow mesenchymal stem cells. We cultivated the bone marrow mesenchymal stem cells in vitro and induced them to be cartilage cells and then mixed cartilage cells with scaffolds. We observed the growth of cartilage cells within the scaffolds by electron microscopy, and then reinjected the mixture in articular cavities of treatment group for one month. Meanwhile, we injected physiological saline into articular cavities of control group and model group. We got the histological grading by evaluating chondral repair results of rabbits. Results Alginate scaffolds had many pores in electron microscopy and these pores were conducive to nutrient movement and cell proliferation. After treatment for one month, cartilage fibrosis alleviated and articular cavity effusion disappeared, which meant cartilage defects were repaired to some extent. Conclusions We can repair cartilage defects of adjuvant-induced arthritis rabbits by injecting the mixture of engineered chondrocytes and alginate scaffolds, and its mechanism may be that scaffolds avail the growth of chondrocytes.

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