关节软骨损伤后体外培养软骨细胞的功能变化

Functional changes in articular chondrocytes cultured in vitro after articular cartilage injury

背景:关节软骨损伤可以影响软骨细胞功能,诱发创伤性骨关节炎。目的:观察关节软骨损伤后体外培养的软骨细胞功能的变化。方法:通过酶消化法分离培养高能量、低能量撞击后和正常兔膝关节透明软骨细胞,观察创伤能量对软骨细胞生存能力的影响;检测软骨细胞合成蛋白多糖和Ⅱ型胶原能力,检测细胞中白细胞介素1β和核转录因子κB mRNA表达水平,检测细胞合成白细胞介素1β和基质金属蛋白酶1的表达。结果与结论:高能量和低能量关节软骨损伤后,软骨细胞的存活率下降,原代细胞的贴壁细胞数量减少,贴壁时间延长,生长曲线下移,细胞甲苯胺蓝染色异染反应减弱,Ⅱ型胶原免疫组化染色强度减弱,软骨细胞中白细胞介素1β和核转录因子κB mRNA表达水平上升,细胞培养液中白细胞介素1β和基质金属蛋白酶1的质量浓度升高,其中高能量组效果更为显著(P<0.05)。说明关节软骨损伤后软骨细胞的功能受到影响,受损程度与创伤强度及炎性细胞因子的表达相关。

BACKGROUND：Articular cartilage injury can influence articular chondrocyte function and induce post-traumatic osteoarthritis. OBJECTIVE：To investigate the functional changes in articular chondrocyte cultured in vitro after articular cartilage injury. METHODS：Rabbit models of articular cartilage injury were established by direct impact of low （0.9 J） and high （6.3 J） energy. Knee joint chondrocytes from normal and articular cartilage injury rabbits were isolated by enzyme digestion to observe the effects of impact on survival capacity of chondrocytes, detect the ability of chondrocytes to synthesize proteoglycan and type Ⅱ collagen, detect intracellular mRNA expression of interleukin-1β and nuclear factor-κB, and measure interleukin-1β and matrix metalloproteinase 1 levels in the culture medium. RESULTS AND CONCLUSION：After high and low energy-induced articular cartilage injury, chondrocytes showed decreased survival rate, primary cells had lower ability of adherence and growth, the intensity of toluidine blue staining and type-Ⅱ collagen immunohistochemical staining were weakened, intracellular mRNA expression of interleukin-1β and nuclear factor-κB was increased,and interleukin-1β and matrix metalloproteinase 1 levels in the culture medium were increased, and these symptoms were more obvious after high energy-induced articular cartilage injury （P 0.05）. These findings suggest that after articular cartilage injury, chondrocyte function is influenced, and the injury degree is related to traumatic intensity and the expression level of inflammatory factors.