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目的 探讨体外培养状态下新生兔软骨细胞的缝隙连接(gap junction)与软骨细胞形态、表型的关系。方法 1.向形成集落的兔膝关节软骨细胞(第5代)培养基内加入荧光染料CFDA-AM,经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。2.以激光淬灭软骨细胞内的荧光后定时测量其荧光的恢复曲线。3.将因股骨颈骨折而行人工关节置换的人股骨头关节软骨切成20um薄片,同样加入荧光染料后测定淬灭前后荧光强度。结果 1.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(范围1-274,n=30),平均值明显低于单个球形软骨细胞者(平均2057,范围340-3538,n=30)。2.只有集落中央的球形软骨细胞才有逐渐的荧光恢复。3.人股骨头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 1.软骨细胞的形态影响其荧光染料的摄入量。2.缝隙连接只出现在球形软骨细胞之间,它与软骨细胞的特有表型同时出现。
Objective To investigate the relationship between the gap junctions of newborn rabbit chondrocytes and the morphology and phenotype of chondrocytes cultured in vitro. Method 1. Fluorescent dye CFDA-AM was added into the culture medium of rabbit knee articular chondrocytes (5th passage) which formed colonies, and the fluorescence intensity of different forms of single chondrocytes was measured by confocal laser scanning microscope. 2. Fluorescence in chondrocytes was laser-quenched to measure the fluorescence recovery curve. 3. Femoral neck fractures due to artificial joint replacement of femoral head articular cartilage cut into 20um sheet, the same fluorescence dye was added before and after quenching the fluorescence intensity. Results 1. The average fluorescence intensity of CFDA-AM in various forms of flat-shaped chondrocytes varied greatly (range 1-274, n = 30), and the mean values were significantly lower than those of single spherical chondrocytes (mean 2057, range 340 -3538, n = 30). Only spherical colony at the center of the colony has a gradual fluorescence recovery. 3. Human femoral head articular cartilage lacunar within the chondrocytes of fluorescence after quenching can be restored by laser. Conclusions 1. The morphology of chondrocytes affects their fluorescent dye uptake. 2. Gap junctions occur only between spherical cartilage cells, which coincide with the unique phenotypes of chondrocytes.