Abstract
The effects of cortisol on basal levels of indices of osteoblast metabolic activity and on PTH regulation of osteoblast activity in vitro were examined in intact bone preparations from neonatal rat calvaria. Uniform punch sections from the frontal portion of calvaria of 3-day-old rats were cultured for 24 h at 37 C in modified BGJb medium. When bone sections were incubated in medium supplemented with cortisol (100 nM) for 24 h, indices of osteoblast metabolic activity, expressed both per total bone section and per micrograms bone DNA, were significantly increased relative to control values. Expressed per micrograms DNA, the following percentage increases were observed in cortisol-treated cultures: alkaline phosphatase activity, +22% (P less than 0.02); [3H] collagen synthesis, +41% (P less than 0.001); and [14C]citrate decarboxylation, + 108% (P less than 0.001). Total DNA per bone section after 24 h was increased by 18% (P less than 0.01), and [3H]thymidine incorporation at 24 h was increased by 26% (P less than 0.01) relative to control values. Stimulation by cortisol occurred in a dose-related manner over concentrations from 1 nM to 1 microM. The stimulatory effects of cortisol were first seen after 6 h of exposure and increased steadily through 24 h of exposure. Incubation in the presence of PTH-(1-34) (100 ng/ml) resulted in significant decrease in alkaline phosphatase activity, collagen synthesis, and citrate decarboxylation after 24 h of exposure (P less than 0.001). The relative order of sensitivity to PTH suppression was identical to the relative sensitivity to cortisol stimulation. In the presence of cortisol (100 nM), the suppressive effect of PTH on all three indices was increased significantly by a factor of 2- to 4-fold. It is concluded that in intact cultured bone, physiological concentrations of cortisol produce both an initial enhancement of indices of osteoblast metabolism and increased osteoblast sensitivity to regulation by PTH.
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