兔膝关节骨性关节炎模型的 建立及软骨细胞的分离、培养

The rabbit knee osteoarthritis model and the separation and culture of cartilage cells

关键词:  骨性关节炎  造模方法  软骨细胞  实验研究

Key words:  osteoarthritis  molding method  cartilage cell  experimental research

摘要:

[目的] 探讨复制兔膝骨性关节炎(KOA)的实验动物模型及其软骨细胞的分离、培养。[方法] 用改良Hulth造模法复制膝骨性关节炎, 进行X线摄片、透射电镜观察造模后细胞形态的改变, 用双酶解法分离膝骨关节软骨细胞, 末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记测定法(TUNEL法)观察细胞的凋亡。[结果] 正常组膝关节面光滑平整, 软骨细胞膜完整。造模组膝关节内侧间隙明显变窄, 关节面粗糙有骨赘生成, 软骨细胞胞核固缩, 细胞膜破损。[结论] 改良Hulth造模法复制KOA操作简单、创伤小, 可保持膝关节一定的稳定性, 为KOA可靠的造模方法。机械-酶消化法可获得数量多、纯度高的软骨细胞, 且软骨的存活率高。

Abstract:

[Objective] To investigate the methods of copying rabbit osteoarthritis of the knee (KOA) model and its isolation, culture of cartilage cells in the experimental animals. [Methods] Using improved Hulth molding method to copy knee osteoarthritis, X-ray examination and transmission electron microscope were used to observe the morphological changing of model cells. The dual enzymatic method was used to separate the cartilage cells, and TUNEL method was used to observe the apoptosis of cells. [Results] Knee joint surface of the normal rabbit was smooth, and the membrane of cartilage cell was integrity. In model group, the knee medial clearance was significantly narrow; the surface of articular was roughness and had osteophyte formation; the cell nucleus of cartilage was pycnosis, and cell membrane was breakage. [Conclusion] Modified Hulth molding method copying KOA is simple, and the operation trauma is small, which can keep the stability of the knee. So KOA is the reliable molding method. Machine-enzyme digestion method may obtain massive cartilage cells with good purity, and the survival rate of cartilage cell is high.