Osteoclasts differentiate from resident precursors in an in vivo model of synchronized resorption: A temporal and spatial study in rats

Osteoclasts differentiate from mononucleated precursors expressing monocyte markers, which gradually evolve to preosteoclasts expressing the osteoclast phenotype, Although the role of osteogenic cells in these changes has been well documented in vitro, their contribution in vivo has not been established, In this study, a synchronized wave of resorption was activated along the mandibular periosteum, The periosteum adjacent to the bone surface studied was separated by a computer-assisted technique into an osteogenic alkaline phosphatase-positive compartment and an outer nonosteogenic compartment. Specific markers (nonspecific esterase [NSE], tartrate-resistant acid phosphatase [TRAP], and EDI antibody, a marker of the monocyte-macrophage lineage) were used to follow osteoclast differentiation quantitatively as a function of time after activation of resorption, from day 0 to day 4 (peak of resorption in this model). Local cell proliferation was assessed in parallel. Between day 0 and day 3, the thickness of the osteogenic compartment decreased by 50% (p < 0.0002), In the osteogenic compartment, proliferating fell numbers fell by 80% at 1/2 day, NSE+ cells (located farthest from the bone surface) increased 3.9-fold on day 4 vs. day 0 (p < 0.005), ED1(+) cells decreased between day 0 and day 2 (p < 0.02) before returning to their initial value, and TRAP(+) cells increased 2.7-fold between day 1 and day 3 (p < 0.0005), Resorption was absent in the site studied on day 0, but on day I there were 20.5 osteoclast nuclei per millimeter of bone surface. The cell ratio changed from 30.3 NSE+ and ED1(+) (some of which were also TRAP(+)) cells per millimeter on day 0 to 37.6 mononucleated cells plus 20.5 osteoclast nuclei on day 4, In the nonosteogenic compartment, an entry of ED1(+)/NSE- was observed on 1/2 day (+23 cells, p < 0.02 vs. day 0). This was followed by a return of ED1(+) cell numbers to the control level on day 1, and a transient increase in NSE+ cells (+47% on day 2 vs. day 1, p < 0.02), TRAP(+) cells were never seen in this compartment. Proliferating cell numbers did not change throughout the study. Our results strongly suggest that the osteoclasts present on day 4 differentiated from the pool of TRAP(+) ED1(+), and NSE+ cells present at the site on day 0, The osteogenic compartment was gradually replenished by cells migrating from the nonosteogenic compartment, which was supplemented by ED1(+) cells recruited from the circulation early after activation. Moreover, osteogenic cells appeared to be as crucial in vivo for the acquisition of the TRAP phenotype as previously shown in vitro. (C) 2000 by Elsevier Science Inc. All rights reserved.