[Regulation of osteoclastic bone resorption. 1. Limitation of osteoclast motile activity by fibroblast-like cells].

The cell-to-cell interaction between osteoclasts and fibroblast-like cells, which were mechanically disaggregated from long bones of neonatal rabbits, was investigated to clarify the cellular mechanisms responsible for the regulation of osteoclastic bone resorption by using phase-contrast time lapse recording and fluorescent microscopy for visualization of F-actin cytoskeleton. Osteoclasts showed rapid extension and retraction of pseudopodia with intense ruffling activity and active translocation over a glass surface when there was enough open space around them, while fibroblast-like cells seemed much more stationary. Osteoclasts were not able to cross over cell bodies of fibroblast-like cells which interfered with migration of the osteoclasts. As fibroblast-like cells proliferated, they gradually took over the open glass surface and finally tightly surrounded the osteoclasts. Such osteoclasts usually became contracted and pseudopodial and migratory activities decreased. Fluorescent microscopy of well-spread osteoclaststs stained with rhodamine-labeled phalloidin revealed development of a band-like structure composed of a number of brightly fluorescent F-actin dots at the cell periphery and fine F-actin bundles which run through the cell bodies. On the other hand, contracted osteoclasts tightly surrounded by fibroblast-like cells showed conglomerates composed of accumulated F-actin dots instead of the band-like structure. An F-actin pattern of fibroblast-like cells, which was very different from that of osteoclasts, displayed thick and straight bright bundles with in the cell bodies. The present observation suggests that ongoing osteoclastic bone resorption may be regulated by fibroblast-like cells through limitation of osteoclastic motile activity, and alteration of cytoskeleton and cell-substratum interaction which occurred in osteoclasts may be involved in the mechanism.