通过骨基质表面培养板检测RANKL诱导破骨细胞骨侵蚀能力

目的探讨以骨基质表面培养板替代骨磨片鉴定破骨细胞骨侵蚀能力的应用方法。方法通过RANKL诱导破骨前体细胞RAW264. 7建立破骨细胞分化模型,运用TRAP染色检测破骨细胞分化程度,并以骨基质表面培养板替代骨磨片行骨陷窝试验检测破骨细胞骨侵蚀能力,以侵蚀面积反映骨侵蚀能力。结果不同浓度的RANKL因子可有效诱导破骨前体细胞RAW264. 7分化为成熟多核破骨细胞,呈浓度依赖性。成熟破骨细胞在骨基质表面培养板上形成不同面积的不规则侵蚀圆环,趋势与破骨细胞分化程度一致。结论使用骨基质表面培养板可有效反映破骨细胞形成及骨侵蚀能力,与TRAP染色结果一致,并且具有操作简便、结果直观、便于统计分析等优点。

Osteo assay surface plate for detection of RANKL-induced mature osteoclast erosion ability

Objective To investigate the application of osteo assay surface plate instead of bone grinding plate to identify the bone erosion ability of osteoclasts. Methods The osteoclast differentiation model was established by RANKL-induced osteoclast precursor cell RAW264.7 cells. The degree of osteoclast differentiation was detected by TRAP staining. The osteo assay surface plate was used instead of the bone grinding plate to test the bone erosion ability of osteoclasts, and the bone erosion ability was reflected as the erosion area. Results Different concentrations of RANKL factor could effectively induce the differentiation of osteoclast precursor cells RAW264.7 into mature multinucleated osteoclasts in a concentration-dependent manner. Mature osteoclasts formed irregular erosion rings of different areas on the surface of osteo assay surface plate, and the trend was consistent with the degree of differentiation of osteoclasts. Conclusion The osteo assay surface plate can effectively reflect the formation of osteoclasts and its bone erosion ability, consistent with the TRAP staining results, and has the advantages of simple operation, intuitive results and convenient statistical analysis.