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目的探讨不同浓度RANKL对骨髓单核细胞向破骨细胞分化和活性的影响。方法 50、100和150μg/L三种RANKL浓度诱导骨髓单核细胞向破骨细胞分化。抗酒石酸酸性磷酸酶染色观察破骨细胞形态并计数,抗酒石酸酸性磷酸酶活性检测试剂盒检测上清中的抗酒石酸酸性磷酸酶活性,骨板吸收实验检测破骨细胞骨吸收活力。结果 100μg/L和150μg/L RANKL浓度组在破骨细胞数量、细胞大小、核数目,抗酒石酸酸性磷酸酶活性和骨吸收率方面均明显高于50μg/L组(P<0.01),而100μg/L和150μg/L组间没有明显差异。结论100μg/L RANKL浓度下破骨细胞形成和活性能力强且相对较经济,是破骨细胞诱导培养的理想浓度值。
Objective To investigate the effects of different concentrations of RANKL on the differentiation and activity of bone marrow mononuclear cells into osteoclasts. Methods Three RANKL concentrations of 50, 100 and 150 μg / L induced differentiation of bone marrow mononuclear cells into osteoclasts. Tartrate-resistant acid phosphatase staining was used to observe the morphology of osteoclasts. The tartrate-resistant acid phosphatase activity assay kit was used to detect the tartrate-resistant acid phosphatase activity in the supernatant. The bone resorption assay was used to detect the bone resorption activity of osteoclasts. Results The levels of osteoclasts, cell size, nucleus number, tartrate-resistant acid phosphatase activity and bone resorption in 100μg / L and 150μg / L RANKL groups were significantly higher than those in 50μg / L group (P <0.01) / L and 150μg / L group no significant difference between groups. Conclusion The ability of osteoclast formation and activity at 100μg / L RANKL is strong and relatively economical, which is an ideal concentration for osteoclast induction culture.