周期性压力对组织工程软骨的影响及其作用机制

The explore of the effects of the cyclic pressure on constructing of tissue engineering cartilage and its mechanisms

目的探讨周期性压力对组织工程软骨的影响及其作用机制。方法取3个月龄新西兰兔关节软骨细胞,培养至第4代后,将软骨细胞随机分为两组。一组为压力组,即关节软骨细胞在周期性压力下培养；另一组为对照组,关节软骨细胞在常规条件下培养。采用组织化学、免疫组织化学观察周期性压力对组织工程软骨的结构以及Ⅱ型胶原分泌的影响。采用激光共聚焦显微镜观察周期性压力对软骨细胞内游离钙离子浓度变化的影响。结果周期性压力下培养的组织工程软骨与常规条件下培养的组织工程软骨相比,支架内软骨细胞数量多、排列规则,Ⅱ型胶原丰富,而且软骨细胞内游离钙离子浓度明显高于对照组。结论周期性压力能够通过增加细胞内游离钙离子浓度来促进软骨细胞的增殖,刺激软骨细胞合成分泌Ⅱ型胶原等细胞外基质。

Objective To explore the effect of the cyclic pressure on the constructing of tissue engineering cartilage in vitro and its mechanisms, Methods Free chondrocytes isolated from rabbit articular cartilage were seeded into PGA scaffolds after expansion in vitro, Experimental group were subjected to the cyclic pressure while the control group were cultured under normal condition. Both groups were cultured in vitro. 5days later, the concentration of calcium within chondrocytes was detected with the use of the laser confocal microscopy. After 4 weeks, the tissue engineering cartilage was observed. The type Ⅱ collagen was assessed by histological staining of HE and immunohistochemistrical stain from the tissue engineering cartilage, Results At the 5th day, the concentration of the calcium of the experiment group was higher than the control group. And after 4 weeks culture, the tissue engineered cartilage under cyclic pressure is morphologically thicker than control group, and shows a higher extra cellular matrix secretion and type Ⅱ collagen synthesis. Conclusion The cyclic pressure promotes chondrocytes to secrete type Ⅱ collagen. And it is good for the construction of tissue engineering cartilage in vitro, the cyclic pressure promote chondrocytes to secrete the type Ⅱ collagen by adjusting the intracellular free calcium concentration.