H+ stimulation of cell-mediated bone resorption in tissue culture.

The addition of protons in the form of hydrochloric acid (10.5, 17.2, or 26.6 meq/l resulting in an initial media pH of 7.28, 7.15, and 6.94, respectively) to neonatal mouse calvaria maintained in a chemically defined medium in tissue culture for 1 wk increased calcium release in a dose-response fashion. The same amounts of protons added to the media of devitalized calvaria caused no increase in calcium release into the medium. The net cell-mediated calcium release resulting from the addition of 26.6 meq/l of protons amounted to approximately 50% of the initial calvarial calcium content. Hydroxyproline determinations revealed that active resorption was taking place, wherein both mineral and organic matrix are removed simultaneously. Histological examination of the extensively resorbed calvaria demonstrated the presence of numerous osteoclasts in different stages of bone destruction. The addition of indomethacin (100 ng/ml) strongly inhibited the increase in calcium release by added protons, suggesting that prostaglandin synthesis is involved in the phenomenon. The addition of thyrocalcitonin also inhibited proton-induced calcium release, providing additional evidence that the calcium release from cultures exposed to added protons involved osteoclastic activity.