转化生长因子β对体外破骨细胞分化的影响

The effect of TGF-β on osteoclast differentiation in vitro

目的:研究在核因子κB受体活化因子配基(RANKL)和巨噬细胞集落刺激因子(M-CSF)联合应用诱导体外骨髓细胞培养系统中,转化生长因子(TGF-β)对破骨细胞分化的影响。方法:选用小鼠M-CSF依赖性非附着性骨髓细胞,在含有25 ng/m l sM-CSF和30 ng/m l sRANKL的α-MEM培养液中进行培养,比较加入和不加入1 ng/m lTGF-β1培养4、9 d后,所形成的抗酒石酸酸性磷酸酶染色(TRAP)阳性多核细胞的数目和骨吸收面积。结果:小鼠骨髓细胞在RANKL和M-CSF共同作用下可形成TRAP阳性多核巨细胞,并具有骨吸收功能。加入TGF-β后,破骨样细胞的数目及骨吸收面积显著增加。结论:TGF-β对RANKL和M-CSF诱导的破骨细胞的分化及其功能的促进作用,可能为炎症状态下发生过度骨吸收的机制之一。

Objective：To investigate the effect of TGF-β on osteoclast differentiation in primary murine bone marrow cell culture in the presence of receptor activator of NF-κB Ligand （RANKL） and macrophage colony stimulating facotr（M-CSF）. Methods：M-CSF-dependent bone marrow cells were isolated from 5 -6 weeks old mice, and cultured in the presence of RANKL at 30 ng/ml and M-CSF at 25 ng/ml with or without I ng/ml of TGF-β for 4 and 9 days respectively. TRAP positive multinucleated cells and resorption pits on dentine slices were counted under light microscope. Results：TRAP positive multinucleated cells were induced from murine bone marrow cell cultures with both RANKL and M-CSF, and these cells formed resorption pits on dentine slices. The number of TRAP positive multinucleated cells and resorption pits were significantly increased by 1 ng/ml of TGF-β（ P 〈0. 01 ）. Conclusion： TGF-β can augment osteoclast formation and function, which may partially account for the bone destruction under inflammation.