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Nitrogen-containing bisphosphonates were shown to cause macrophage apoptosis by inhibiting enzymes in the biosynthetic pathway leading from mevalonate to cholesterol. This study suggests that, in osteoclasts, geranylgeranyl diphosphate, the substrate for prenylation of most GTP binding proteins, is likely to be the crucial intermediate affected by these bisphosphonates. We report that murine osteoclast formation in culture is inhibited by both lovastatin, an inhibitor of hydroxymethylglutaryl CoA reductase, and alendronate. Lovastatin effects are blocked fully by mevalonate and less effectively by geranylgeraniol whereas alendronate effects are blocked partially by mevalonate and more effectively by geranylgeraniol. Alendronate inhibition of bone resorption in mouse calvaria also is blocked by mevalonate whereas clodronate inhibition is not. Furthermore, rabbit osteoclast formation and activity also are inhibited by lovastatin and alendronate. The lovastatin effects are prevented by mevalonate or geranylgeraniol, and alendronate effects are prevented by geranylgeraniol. Farnesol and squalene are without effect. Signaling studies show that lovastatin and alendronate activate in purified osteoclasts a 34-kDa kinase. Lovastatin-mediated activation is blocked by mevalonate and geranylgeraniol whereas alendronate activation is blocked by geranylgeraniol. Together, these findings support the hypothesis that alendronate, acting directly on osteoclasts, inhibits a rate-limiting step in the cholesterol biosynthesis pathway, essential for osteoclast function. This inhibition is prevented by exogenous geranylgeraniol, probably required for prenylation of GTP binding proteins that control cytoskeletal reorganization, vesicular fusion, and apoptosis, processes involved in osteoclast activation and survival.

The vast majority of total hip prostheses currently implanted consist of a hard metal or ceramic femoral head articulating against an ultra-high molecular weight polyethylene (UHMWPE) acetabular cup. Over the last 10 years, evidence has accumulated to show that these prostheses are prone to failure due to late aseptic loosening and few survive beyond 25 years. With an increasing need to implant hip prostheses in the younger, more active patient the need to understand the mechanisms of failure and to develop artificial hip joints using alternative materials have become major issues in the orthopaedic community. This review focuses initially on our current understanding of the biological reactions to UHMWPE prosthetic wear debris in vivo and in vitro since this is believed to be the main cause of late aseptic loosening. While the precise mechanisms of osteolysis induced by UHMWPE wear debris have not been elucidated, the major message to emerge is that it is not the wear volume that determines the biological response to the debris, but the concentration of the wear volume that is within the critical size range (0.2-0.8 micron) for macrophage activation. The review then considers whether the problem of wear-debris-induced osteolysis may be overcome with the use of new generation metal-on-metal or ceramic-on-ceramic prostheses. For metal-on-metal prostheses, the prospects for increasing the osteolysis free life of the implant are good but additional biological problems associated with the nanometre size and reactivity of the wear particles in vivo may emerge. For the ceramic-on-ceramic prostheses, although initial prospects are encouraging, more data are needed on the characteristics of the wear particles generated in vivo before predictions can be made. It is concluded that the pre-clinical testing of any new materials for joint replacement must include an analysis of the wear particle characteristics and their biological reactivity in addition to the usual assessment of wear.

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Friction of cartilage on metal, metal on cartilage and cartilage on cartilage contact configurations, within a mixed lubrication regime, was measured using synovial fluid, Ringer's solution or with no lubricant present. The main test variable was the period of stationary loading which ranged from 5 s to 45 min, prior to sliding and consequently measuring friction. The coefficient of friction rose gradually with increasing stationary loading time, up to a value of approximately 0.3 at 45 min for all the contact configurations. Following the re-application of load, after short periods of load removal, friction was also found to drop sharply. The flow of liquid in the biphasic cartilage and load carriage by the fluid phase was highlighted as being an important factor in reducing friction within the mixed or boundary lubrication regime. Movement of the contact zone over the cartilage counterface ensured very low friction as the slider moved over fully hydrated cartilage. For the cartilage--cartilage contacts synovial fluid significantly reduced friction compared to Ringer's solution. This was attributed to an effective boundary lubrication action, which was not as effective for the cartilage--metal contacts.

Multinucleated bone-resorbing osteoclasts (Ocl) are cells of hematopoietic origin that play a major role in osteoporosis pathophysiology. Ocl survival and activity require M-CSF and RANK ligand (RANKL). M-CSF signals to Akt, while RANKL, like TNFa, activates NF-jB. We show here that although these are separate pathways in the Ocl, signaling of all three cytokines converges on mammalian target of rapamycin (mTOR) as part of their antiapoptotic action. Accordingly, rapamycin blocks M-CSF-and RANKL-dependent Ocl survival inducing apoptosis, and suppresses in vitro bone resorption proportional to the reduction in Ocl number. The cytokine signaling intermediates for mTOR/ribosomal protein S6 kinase (S6K) activation include phosphatidylinositol-3 kinase, Akt, Erks and geranylgeranylated proteins. Inhibitors of these intermediates suppress cytokine activation of S6K and induce Ocl apoptosis. mTOR regulates protein translation acting via S6K, 4E-BP1 and S6. We find that inhibition of translation by other mechanisms also induces Ocl apoptosis, demonstrating that Ocl survival is highly sensitive to continuous de novo protein synthesis. This study thus identifies mTOR/S6K as an essential signaling pathway engaged in the stimulation of cell survival in osteoclasts.

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