OPGL诱导小鼠外周血单核细胞分化为破骨细胞及其破骨活性

目的 :观察在小鼠外周血单核细胞培养中破骨细胞抑制因子配基 (osteoprotegerinligand ,OPGL)诱导单核细胞分化为破骨细胞的作用 ,以及地塞米松对破骨细胞分化和骨吸收活性的影响 .方法 :将外周血单核细胞分组培养 ,在A组中加入巨噬细胞集落刺激因子和OPGL ,B组中再加入地塞米松 .多核巨细胞形成后行抗酒石酸酸性磷酸酶 (tartratere sistantacidphosphatase,TRAP)及甲苯胺蓝染色 .将象牙骨片上骨吸收面积经计算机图像分析以确定其差异 .结果 :外周血单核细胞培养 7～ 10d ,可以见到大量的多核巨细胞 ,体积巨大 .B组中 ,多核巨细胞出现较A组早 .几乎所有的多核巨细胞表现为TRAP强阳性 ,而多数单核细胞和巨噬细胞为TRAP阴性或弱阳性 .在象牙骨片上有大量的骨吸收陷窝形成 ,A、B两组的骨吸收无明显差异 (P >0 .0 5 ) .结论 :OPGL可以诱导小鼠外周血单核细胞分化为破骨细胞并具有骨吸收活性 .地塞米松可加速OPGL诱导的破骨细胞分化作用 ,但对于破骨细胞的骨吸收活性无影响

Observation of the formation and function of osteoclasts derived from mouse peripheral blood monocytes induced by osteoprotegerin ligand

AIM: To observe the induction of osteoprotegerin ligand in the differentiation and function of osteoclasts derived from mouse peripheral blood monocytes (PBMCs). METHODS: After the mouse PBMCs were isolated and cultured in wells, which contained glass coverslips and ivory dentine slices, macrophage colony stimulating factor (M CSF) and osteoprotegerin ligand (OPGL) were added. Dexamethasone was also added in the wells of Group B but not Group A. When the multinuclear cells were observed in the culture, the coverslips were stopped and stained for tartrate resistant acid phosphatase (TRAP). Seven days after the stop of coverslips, the dentine slices were stopped and stained for toluidine blue to observe bone resorption by osteoclasts, and the areas of bone resorption were analyzed by computer. RESULTS: After 7 or 10 days culture of PBMCs, plenty of large multinuclear cells could be found on the coverslips and in the wells of Group B, the formation of multinuclear cells was seen earlier than Group A. Most of multinuclear cells were TRAP positive and most of monocytes and macrophages were negative or weakly positive. Bone resorption was observed on the dentine slices and the statistic analysis did not show any significant difference between different groups ( P >0.05). CONCLUSION: Cultured with OPGL, mouse PBMCs can differentiate into osteoclasts and induce bone resorption on dentine slices. It seems that dexamethasone can accelerate the formation of osteoclasts but it has no effects on bone resorption by osteoclasts.