软骨细胞形态、表型与胞间通讯研究

SHAPE,PHENOTYPE AND GAP JUNCTION OF THE RABBIT CHONDROCYTES

目的 探讨体外培养下新生兔软骨细胞的缝隙连接与软骨细胞形态、表型的关系。方法 向 96孔板内的第 5代新生兔膝关节软骨细胞培养基内加入荧光染料 CFDA- AM,激光共聚焦显微镜测量不同形态的单个软骨细胞内的荧光强度 ;以激光淬灭软骨细胞内的荧光后定时测量荧光恢复曲线。将因股骨颈骨折行人工关节置换的人股骨头关节软骨切成 2 0μm薄片 ,同样加入荧光染料后测定激光淬灭前后荧光强度。结果 各种形态的扁平状软骨细胞摄入CFDA- AM后平均荧光强度差异很大平均 83(1～ 2 74 ) ,平均值明显低于单个球形软骨细胞者平均 2 0 5 7(340～ 35 38) ;只有集落中央的球形软骨细胞才有逐渐的荧光恢复 ;但关节软骨陷窝内软骨细胞内的荧光经激光淬灭后可以恢复。结论 软骨细胞的形态影响其荧光染料的摄入量 ;缝隙连接只出现在球形软骨细胞之间 ,与软骨细胞的特有表型同时出现 ;股骨头人关节软骨陷窝内的软骨细胞间有缝隙连接存在 。

Objective To study the gap junction and phenotype of cultured chondrocyte of rabbit ,and the gap junction between the chondrocytes in the same cartilage cavities in human femoral head articular cartilage. Methods CFDA AM was added into the medium of the fifth passage of chondrocyte of rabbit in the 96 well plate. The fluorescent in spherical and fibroblast like chondrocytes was detected separately. The recurrence of the fluorescent in accordant with time in 16 minutes was recorded after blanching the fluorescent with laser. And the fluorescent after blanching of chondrocyte in the cartilage cavities in the proliferative zone of articular cartilage of adult human femoral head was recorded, too. Results The average fluorescent of the single layer of the fibroblast like chondrocyte was 83(ranged from 1 to 274), the highest was found in the spherical shaped cell (averaged 2 057, ranged from 340 to 3 538). The recurrence of the fluorescent after the blanching appeared only in the spherical chondrocyte, the gap junctions reappeared only in the spherical chondrocytes, as well as in the cells in the cartilage cavities in the articular cartilage of the human femoral head. Conclusion The appearance of the gap junction is corresponded with the spherical shape, secretion of the cartilage matrix of the chondrocyte. There are gap junctions in the cells in the same cartilage cavities in the articular cartilage of the human femoral head, while no gap junctions in the isolated chondrocytes in the cartilage.