组织块法与传统破骨细胞培养方法对骨吸收功能的比较研究

The Comparison of Osteoclast Bone Resorption for Tissue Mass Culture Method and Traditional Culture Method

目的:比较组织块法与传统破骨细胞培养方法对破骨细胞骨吸收功能的影响。方法:用组织块法与传统破骨细胞培养方法进行破骨细胞培养,抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色鉴定破骨细胞,超声去除骨片上细胞后,骨片经1%甲苯胺蓝染色,光镜下观察骨片上吸收陷窝。根据骨片上陷窝数目、面积以及深度定量比较两组破骨细胞骨吸收功能。结果:吸收陷窝经甲苯胺蓝染色后光镜下可清晰识别,组织块法在骨片上的吸收陷窝数目比传统法明显少,而且骨片上陷窝面积及深度也比传统法小而浅(P<0.05)。结论:组织块法培养的破骨细胞无论从数量上还是从功能上均没有传统法培养的好。数量大的成骨细胞对共培养破骨细胞骨吸收功能起到负面作用。

Objective：To compare osteoclast bone resorption function between the tissue mass culture method and the traditional culture method.Methods：The tissue mass culture method and the traditional culture method were used to culture osteoclasts,tartrate-resistant acid phosphatase（TRAP） staining was used to identify osteoclasts,the cells on the bone chip were removed by ultrasonic,the bone chips were stained with 1% toluidine blue for 3~4min,absorb lacunaes were observed by light microscope on the whole chip.According to the number of absorb lacunae,the area of lacunae and lacunae depth quantitative were compared in bone resorption functions of two groups.Results：The absorption lacuna staining by toluidine blue in light microscopy had been clearly identified,the absorption lacuna number of tissue mass culture method in bone chip was less than that of the traditional culture method,and the lacunae size and depth in bone chip were smaller and shallower than that of the traditional method（P0.05）.Conclusion：In terms of quantity and function,the osteoclasts for tissue mass cultured method are not well than that of traditional cultured methods.Larger number osteoblasts co-culture with osteoclast plays a negative role in the bone resorption for osteoclasts.