Chambers T J, Revell P A, Fuller K, Athanasou N A
J Cell Sci. 1984 Mar;66:383-99. doi: 10.1242/jcs.66.1.383.
A carborundum wheel was used to prepare slices of cortical bone that demonstrate a predictable surface appearance in the scanning electron microscope. Osteoclasts were mechanically disaggregated from neonatal rabbit long bones and settled onto these slices. After 24h in culture osteoclasts were associated with areas of excavation in the bone surface. These excavated areas typically showed a well-defined outline and a distinctive fibrillar base, which resembled the pattern of collagen fibrils in bone. The majority of such concavities were of approximately circular outline and of smaller diameter than the associated osteoclast, but other excavations were elongated or of complex morphology, and may have been produced by osteoclasts that were resorbing bone while they migrated. Irregular concavities tended to be more shallow but to occupy a greater area of the bone surface than circular concavities. Roughening of the bone surface without detectable excavation was also seen adjacent to osteoclasts. Calcitonin and cytochalasin B, which inhibit osteoclastic motility, also inhibited bone resorptive activity by these cells. The techniques described in this paper represent a model system with which to assess the direct and indirect effects of hormones, cells and substrate composition on the induction, stimulation and inhibition of osteoclastic bone resorption and to investigate the mechanisms by which cells degrade extracellular matrices.
用碳化硅砂轮制备皮质骨切片,这些切片在扫描电子显微镜下呈现出可预测的表面外观。破骨细胞从新生兔长骨中机械分离出来并附着在这些切片上。培养24小时后,破骨细胞与骨表面的侵蚀区域相关联。这些侵蚀区域通常显示出轮廓清晰且有独特的纤维状基底,类似于骨中胶原纤维的模式。大多数此类凹陷轮廓近似圆形,直径小于相关的破骨细胞,但其他侵蚀区域呈细长形或形态复杂,可能是由迁移时正在吸收骨的破骨细胞产生的。不规则凹陷往往较浅,但比圆形凹陷占据更大的骨表面积。在破骨细胞附近还可见到骨表面粗糙但无明显侵蚀的情况。抑制破骨细胞运动的降钙素和细胞松弛素B也抑制了这些细胞的骨吸收活性。本文所述技术代表了一个模型系统,可用于评估激素、细胞和底物成分对破骨细胞性骨吸收的诱导、刺激和抑制的直接和间接影响,并研究细胞降解细胞外基质的机制。