颞下颌关节骨关节病髁状突软骨细胞体外培养及细胞生物学特性研究

In vitro culture of mandibular condylar chondrocytes obtained from the rabbit model of temporomandibular joint osteoarthrosis and the study of its cell biology characteristics

目的 :从兔的颞下颌关节骨关节病模型动物获得髁状突软骨细胞并行体外培养。方法 :采用部分关节盘手术切除的方法制造颞下颌关节骨关节病模型 ,组织大体观察、超微结构观察和组化检测方法确定骨关节病的存在。在此基础上 ,通过机械分离、胰蛋白酶和胶原酶联合消化的方法从骨关节病髁状突软骨组织中获得软骨细胞 ,进行体外环境下的贴壁培养 ;通过软骨细胞特异性蛋白的免疫组化方法进行细胞鉴定。四甲基偶氮唑盐法比较病理模型细胞与正常细胞的增殖能力差异。结果 :上述颞下颌关节骨关节病模型全面地模拟了骨关节病变关节软骨组织的特征 ;利用机械分离、胰蛋白酶胶原酶联合消化的方法成功地从骨关节病髁状突软骨组织中分离出软骨细胞并在体外贴壁条件下培养成活 ;应用特异性的Ⅱ型胶原多克隆抗体和聚合性糖胺多糖单克隆抗体对培养细胞进行免疫组化检测鉴定 ,均显示阳性。骨关节病髁状突软骨细胞的增殖能力高于正常细胞。结论 :成功地建立了骨关节病髁状突软骨细胞的体外模型 ;骨关节病早期的髁状突软骨细胞表现出增殖活跃的特性。

Objective:To culture the mandibular condylar chondrocytes from the rabbit model of osetoarthrosis of temporomandibular joint(TMJ) in vitro and investigate their biological features. Methods:TMJ osteoarthrosis model was made by the partial resection of joint disc. The diagnosis was confirmed by means of macroscopy, ultrostructure observation and histochemical examination. The chondrocytes were harvested from osteoarthritic condylar cartilage by mechanical dissection and digestion with trypsin and collagenase, and then cultured in vitro under the monolayer culture condition. Identification of primary cultured cells was made histoimmunologically with type Ⅱ collagen antibody and aggrecan antibody. The proliferative potential of osteoarthritc chondrocytes was studied to outline the basic feature of the cells.Results:The TMJ osteoarthrosis model used in this study could effectively reflect the pathological feature of osteoarthritic cartilage. The chondrocytes harvested from osteoarthritic condylar cartilage by mechanical dissection and enzyme digestion grew well in vitro under the monolayer culture condition. The positive results of histoimmunological examination with cartilage-specific protein, type Ⅱ collagen and aggrecan, confirmed that the culture cells were chondrocytes. The proliferative potential of the osteoarthritic chondrocytes is much higher than that of the normal control. Conclusion:The chondrocytes of the rabbit TMJ osteoarthrosis model could be harvested by using the method suggested by the present study. The chondrocytes in the early stage of osetoarthrosis showed a rather active proliferative potential.