Calcium signalling via multiple P2 purinoceptor subtypes in rat osteoclasts.

Extracellular nucleotides bind to P2 purinoceptors in many tissues. P2X purinoceptors are intrinsic ion channels that mediate depolarization and influx of Ca(2+), whereas P2Y purinoceptors are coupled through G-proteins to mobilization of intracellular Ca(2+). Previous studies have yielded conflicting information on the responses of osteoclasts to nucleotides. The purpose of this study was to investigate the pathways underlying purinoceptor-mediated Ca(2+) signalling in authentic mammalian osteoclasts. Osteoclasts, isolated from the long bones of neonatal rats, were loaded with the Ca(2+)-sensitive probe fura-2 and [Ca(2+)](i) was monitored by microspectrofluorimetry. ATP (10-100 microM) induced transient elevation of [Ca(2+)](i) in 74% of osteoclasts tested. Similar responses were observed in Ca(2+)-free media, consistent with release of Ca(2+) from intracellular stores. Oscillations in [Ca(2+)](i) were observed only in osteoclasts that had a 'rounded' morphology. Responses to selective P2 agonists were consistent with the presence of multiple purinoceptor subtypes, including members of both the P2Y and P2X families. Alendronate, a bisphosphonate with structural similarities to methylene ATP analogues, neither activated nor blocked the Ca(2+) response mediated by osteoclast purinoceptors. Mechanical stimulation of osteoclasts elicited transient elevation of [Ca(2+)](i) which involved Ca(2+) influx and, in some cases, release from stores. The nucleotidase apyrase did not inhibit deformation-induced elevation of [Ca(2+)](i) in the presence of extracellular Ca(2+), indicating that nucleotide release is not essential for mechanically induced Ca(2+) influx. These findings indicate that osteoclasts exhibit multiple P2 purinoceptor subtypes, linked to elevation of [Ca(2+)](i).

Osteoclast activation in inflammatory periodontal diseases.

OBJECTIVE: In this paper, we review the mechanisms thought to be involved in the activation of osteoclasts in periodontitis. SUMMARY: Osteoclasts are regulated by both microbial and host factors. Some factors act directly on cells of the osteoclast lineage, whereas others act indirectly through other cell types in the bone environment. The proinflammatory cytokines (interleukins 1 and 6, tumor necrosis factors) have been implicated in the stimulation of osteoclastic resorption. The roles of the immunoregulatory cytoknes (interleukins 2 and 4, interferon gamma) are less clear, but decreased levels of these factors may contribute to periodontitis. A number of lipid mediators may be involved in stimulation of bone resorption. These include bacterial lipopolysaccharide and host-derived platelet-activating factor and prostaglandins. More recently, reactive oxygen intermediates and extracellular nucleotides, both present at sites of inflammation, have been investigated as possible modulators of osteoclast activity. The potential use of antiresorptive therapies in periodontitis is reviewed. CONCLUSIONS: A wide range of host and bacterial factors contribute to the loss of alveolar bone in periodontitis. However, much remains to be understood about the complex mechanisms through which these factors regulate osteoclast activity. Further studies at the cellular and molecular level will lead to a better understanding of these processes and perhaps suggest new approaches for periodontal therapy.