Domon T, Wakita M
Department of Oral Anatomy II, Hokkaido University School of Dentistry, Sapporo, Japan.
Am J Anat. 1991 Sep;192(1):35-44. doi: 10.1002/aja.1001920105.
Osteoclasts collected from the long bones of mice were cultured on dentin slices. To identify osteoclasts, the tartrate-resistant acid phosphatase (TRACPase) activity of cultured cells was histochemically examined by the azo dye method. The TRACPase-positive cells could be distinguished from other cells by light microscopy. The cells were sectioned by alternating semithin and ultrathin sections to observe their ultrastructure and three-dimensional structure. TRACPase activity was detected both in multi-nucleated osteoclasts and in mononuclear cells. Most of the mononuclear TRACPase-positive cells had features similar to preosteoclasts. A mononuclear TRACPase-positive cell was a ruffled border and clear zone was reconstructed three-dimensionally by NIKON COSMOZONE 2SA. The reconstruction showed that this cell possessed a large clear zone and small ruffled border. Under the ruffled border, no lacuna was apparent; but there was disruption of the dentin surface. The results suggest that this cell was a mononuclear osteoclast and that it might have been in the process of making a new lacuna.
从小鼠长骨采集的破骨细胞在牙本质切片上进行培养。为鉴定破骨细胞,采用偶氮染料法对培养细胞的抗酒石酸酸性磷酸酶(TRACPase)活性进行组织化学检测。通过光学显微镜可将TRACPase阳性细胞与其他细胞区分开来。通过交替制作半薄切片和超薄切片对细胞进行切片,以观察其超微结构和三维结构。在多核破骨细胞和单核细胞中均检测到TRACPase活性。大多数单核TRACPase阳性细胞具有类似于破骨前体细胞的特征。一个单核TRACPase阳性细胞的皱褶缘和透明区通过尼康COSMOZONE 2SA进行三维重建。重建显示该细胞具有大的透明区和小的皱褶缘。在皱褶缘下方,未见明显的陷窝;但牙本质表面有破坏。结果表明该细胞是单核破骨细胞,可能正处于形成新陷窝的过程中。