Objective To investigate whether the lyophilized powder of mycobacterium tuberculosis dealt with ultrasonic spallation or heat spallation can induce osteoclasts formation from mouse bone marrow mononuclear phagocytes,and to discuss the role of TNF-α in the mechanism of osteoclastogenesis in osteoarticular tuberculosis.Methods The system of osteoclasts vitro culture was established.The lyophilized powder at different concentrations of mycobacterium tuberculosis dealt with ultrasonic spallation or heat spallation were added to the cell supematant of the osteoclast system in vitro culture,and their effects on the induction of osteoclasts formation from mouse bone marrow mononuclear phagocytes were observed.Collecting cell supematant after incubation 24 h and 48 h,and the concentration of TNF-α in cell supernatant was detected by ELISA.The TRAP staining and bone resorption experiments were used to identify the functions of osteoclasts.Results The lyophilized powder of mycobacterium tuberculosis dealt with heat spallation could not stimulate mouse bone marrow mononuclear cells to secrete TNF-α,could not induce cell differentiation of mouse bone marrow mononuclear cells into osteoclasts,and it also could not promote bone destruction.While the lyophilized powder of mycobacterium tuberculosis dealt with ultrasonic spallation stimulated bone marrow mononuclear cells to secrete TNF-α,induced cell differentiation of mouse bone marrow mononuclear cells into osteoclasts,and it also led to bone destruction.Conclusion The lyophilized powder of mycobacterium tuberculosis dealt with ultrasonic spallation can induce cell differentiation of mouse bone marrow mononuclear cells into osteoclasts,and it also can lead to bone destruction.Meanwhile,the osteoclastogenesis and osteoclasts activation have a certain correlation with the TNF-α concentration value.%目的 观察热处理和超声裂解结核杆菌(MTB)冻干物能否诱导小鼠骨髓单核细胞形成破骨细胞,探讨培养液中TNF-α水平与诱导破骨细胞形成的相关性及其机制.方法 建立体外破骨细胞培养体系,观察不同浓度热处理MTB冻干物和超声裂解MTB冻干物对体外诱导小鼠骨髓单核细胞形成破骨细胞的影响.收集诱导24、48h的细胞培养液,采用ELISA法定量检测细胞培养液中的TNF.-α浓度,TRAP染色及骨吸收陷窝实验鉴别破骨细胞功能.结果 热处理MTB冻干物不能刺激骨髓单核细胞分泌TNF-α,也不能诱导骨髓单核细胞形成破骨细胞和促进骨质破坏；超声裂解MTB冻干物能刺激骨髓细胞分泌TNF-α,且能促进破骨细胞形成,导致骨质破坏.结论 超声裂解MTB冻干物能够诱导小鼠骨髓单核细胞形成破骨细胞,导致骨质破坏,且破骨细胞的形成及活化与TNF-α水平相关.
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