No abstract

Extracts of tumors from patients with humoral hypercalcemia of malignancy (HHM) were tested using an in vitro bone resorption assay in order to investigate the pathogenesis of the hypercalcemia. Bone resorption was assessed by comparing the percent release of previously incorporated 45Ca from paired halves of newborn mouse calvaria. Saline extracts of three out of five tumors from HHM patients caused a significant increase in 45Ca release relative to controls. Extracts of liver and non-HHM tumor did not cause significant resorption. Tumor-stimulated bone resorption was blocked by indomethacin and eicosatetraynoic acid, inhibitors of the synthesis of prostaglandins (PGs) and related metabolites of arachidonic acid, whereas resorption stimulated by parathyroid hormone (PTH), PGE2, or 1,25-(OH)2D3 was not. Furthermore, levels of immunoreactive PTH or PGE2 in tumor extracts were not sufficient to account for the degree of resorption observed. These observations indicate that PTH or PGE2 are not responsible for the bone resorption caused by extracts of tumors from these patients with HHM. Furthermore, they suggest that hypercalcemia in these patients may result from bone resorption stimulated by the local production in bone of PGs or related metabolites of arachidonic acid in response to a humoral factor elaborated by the tumor.

Mononuclear phagocytes have been implicated as important cellular elements in the process of bone resorption. We have postulated that the recruitment and migration of mononuclear phagocytes to bone occurs via a mechanism(s) in which bone-derived chemotactic factors (BDCF) are released from foci undergoing resorption. In the experiments presented here we have used newborn mouse calvaria and examined a variety of extraction protocols, both dissociative and non-dissociative, as means of obtaining stable and reproducible chemotactic activity for mouse peritoneal macrophages. Chemotaxis and chemokinesis were assessed using a multi-well chamber modification of the Boyden transfilter method. Further, we have attempted to purify the BDCF by both molecular sieve and anion exchange chromatography. Our results indicated that non-dissociative extraction with 0.5 M EDTA in the presence of 1% DMSO yielded the most potent and reproducible chemotactic activity. The results of molecular sieve and anion exchange chromatography suggested that there were several BDCF activities in these preparations and that their molecular weights were probably in the range of from 14,000-67,000 daltons. Anion exchange chromatography also demonstrated the presence of a fraction, eluted with 2 M NaCl, with high chemotactic activity and minimal protein concentration. These observations confirmed the suggestion that there are several macrophage chemotactic factors in bone which have as yet to be identified, and suggest methods for pursuing their isolation.

No abstract

The mechanisms that control cycles of bone formation and bone resorption are not well understood. In this report we provide evidence that compound 48/80 is a potent inhibitor of bone resorption in vitro. Resorption was assessed by the release of calcium-45 from pre-labelled newborn mouse calvaria that were treated with compound 48/80 and/or parathyroid hormone (PTH) in organ culture. Our results demonstrate that co-incubation of calvaria with PTH plus compound 48/80 (concentrations 1-10 mcg/ml) produces a marked reduction of calcium-45 release compared to PTH alone. Furthermore, pre-incubation of calvaria with compound 48/80, for as little as three hours, inhibits resorption by subsequent treatment with PTH alone. Measurement of lactate dehydrogenase (LDH) released into the culture medium indicated that treatment with compound 48/80, at the doses and time periods studied, was not cytotoxic. This novel effect of compound 48/80 may provide a useful tool for studying the cellular mechanisms involved in the bone resorption process.