持续压力对体外培养的人破骨细胞形态和基质金属蛋白酶9、抗酒石酸酸性磷酸酶表达的影响

目的:探讨施加持续压力对体外培养的人破骨细胞功能的影响.方法:从脐血中提取单核细胞,α-MEM培养液中加入RANKL和M-CSF作为诱导因子,细胞培养14d.对鉴定为阳性的成熟破骨细胞施加100kPa持续性压力,加载时间分别为1h、3h、5h,观察加力后细胞形态的变化;对破骨细胞的2种重要功能酶-基质金属蛋白酶9(MMP-9)和抗酒石酸酸性磷酸酶(TRAP)进行染色,每组随机选取20个细胞进行灰度分析,应用SPSS12.0软件包对MMP-9和TRAP表达量进行t检验,分析持续压力对破骨细胞的影响.结果:在持续压力的作用下,破骨细胞形态由不规则形态向圆形变化发展.加力1h后,TRAP表达量(59.61±14.95)与未加力组(80.01±9.69)有显著性差异(P<0.05);MMP-9在加力5h后(60.44±8.49)表达量增加,有统计学意义(P<0.05).结论:体外持续压力可刺激成熟破骨细胞形态发生变化,MMP-9和TRAP表达量增加,说明应力能上调破骨细胞的骨吸收功能.

The effect of continuously compressive press on the shape of cells and expressions of MMP-9,TRAP in human osteoclasts

PURPOSE: To investigate the effect of continuously compressive pressure on the function of human osteoclasts. METHODS: Monocytes were collected from umbilical cord cells, and were cultured for 14 days. alpha-MEM which contains RANKL and M-CSF was used as the culture medium. 100kPa compressive pressure was applied to the mature osteoclasts for 1 hour,2 hours and 3 hours. The shape change was observed. TRAP,MMP-9 enzymes were stained, and measured randomly. The data were analyzed with SPSS12.0 software package for Student's t test. RESULTS: Compressive pressure made the shape of osteoclasts incline to round. After adding pressure for 1 hour, there was significant difference on the expression of TRAP between pressure group(59.61+/-14.95)and non-pressure group(80.01+/-9.69) (P<0.05). Significant increase expression of MMP-9 occured after 5 hours (P<0.05). CONCLUSION: The changes of shape and levels of functional enzymes show that compressive pressure enhances the bone-absorbing activity. Supported by National Natural Science Foundation of China (Grant No.30471911).