Electron microscopic and histochemical studies of the mononuclear osteoclast of the mouse

Abstract

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.

Ultrastructural characterization of preosteoclasts derived from bone marrow progenitors stimulated by osteoclast colony stimulating factor

“…Abundant lysosomal vacuoles were also observed in mononuclear osteoclast precursor cells (Baron et al, 1986;Domon and Wakita, 1991). Furthermore, coated pits and coated patches described in mononuclear precursors of osteoclasts in a rat model (Baron et al, 1986) were also observed in our cells.…”

“…Our cells cultured with 0-CSF showed a striking resemblance to cells described by Domon and Wakita (1991) in their studies of cultured bone marrow mononuclear cells. The TRAPase-positive cells in our study, however, were derived from bone marrow progenitors that had the capacity to proliferate and form colonies in response to 0-CSF after 14-day incubation; thus, they must be derived from much earlier precursors than the preosteoclasts studied by Domon and Wakita in their 3-day culture experiments.…”

“…Although a number of studies have defined light and electron microscopic features of mature functional osteoclasts in vivo and in vitro, (Lucht, 1972a,b;Holtrop and King, 1977;Jones et al, 1985;Fukushima et al, 1991;McKee and Nanci, 1993), morphological features of immature osteoclasts in the bone marrow are less characterized, with only a few studies found in the literature (Baron et al, 1986;Domon and Wakita, 1991). Osteoclasts play essential roles in bone resorption.…”

Odontoclasts in the Chinook salmon differ from mammalian odontoclasts by exhibiting a great proportion of cells with high nuclei number

“…These reports disregarded the presence of mononuclear osteoclasts which were observed in serial sections based on criteria of both acid phosphatase activity and resorptive lacunae (Kaye, 1984) or odontoclasts because their presence was not proven precisely. Previously, we have demonstrated the presence of mononuclear osteoclasts with ruffled borders in vitro (Domon and Wakita, 1991) and mononuclear odontoclasts in vivo (Domon et al, 1994), and investigated the distribution of number of nuclei per human odontoclast, and reported that 93.8% of cells had 10 or fewer, including mononucleus (Domon et al, 1997). These reports indicate that the distribution of nuclei in osteoclasts or odontoclasts varies little in mammals.…”

Culturing of cells from giant cell tumour of bone on natural and synthetic calcified substrata: the effect of leukaemia inhibitory factor and vitamin D3 on the resorbing activity of osteoclast-like cells

“…Since the number of multinucleated cells was similar in control and experimental 7-day cultures, the effects of both substances were very likely to increase the intrinsic cellular resorption activity. We cannot however, exclude the possibility that VD3 and HILDA/LIF increased the number of mononuclear TRAP-positive cells which possess osteoclastic activity [12,34]. The scoring of these cells in 7-day cultures was very difficult.…”