Cortisol Enhances the Anabolic Effects of Insulin-Like Growth Factor I on Collagen Synthesis and Procollagen Messenger Ribonucleic Acid Levels in Cultured 21-Day Fetal Rat Calvariae*

Abstract

We examined the ability of cortisol to modulate the stimulatory effects of recombinant human insulin-like growth factor-I (IGF-I) on collagen synthesis, procollagen messenger RNA (mRNA) levels and DNA synthesis in 21-day fetal rat calvariae maintained in serum-free organ culture for 24-96 h. Collagen synthesis was quantitated by measuring the incorporation of [3H]proline into collagenase-digestible protein (CDP) and alpha-1(I) procollagen mRNA transcripts were assessed by Northern blot analysis. Cell replication was quantitated by measuring the incorporation of [3H]thymidine into bone. As described previously, 100 nM cortisol had a biphasic effect on CDP labeling, increasing CDP after 24 h and decreasing CDP after 48, 72, and 96 h of culture. IGF-I alone increased CDP labeling by 1.6-fold after 24 h and by 2-fold after 48 or 72 h of culture, and cortisol potentiated this anabolic effect. In the presence of 100 nM cortisol, IGF-I increased CDP labeling by 2.6-fold after 24 h, by 5-fold after 48 h, and by 8-fold after 72 h of culture. A higher concentration of cortisol (1000 nM) also potentiated the IGF-I response on CDP labeling after 96 h of culture. In the presence of 100 nM cortisol, concentrations of IGF-I lower than 10 nM consistently increased CDP labeling and the percent collagen synthesized whereas these concentrations were not always effective without cortisol. PTH, which like cortisol decreased basal CDP labeling, did not enhance the stimulatory effects of IGF-I. Cortisol also enhance the stimulatory effects of IGF-I on alpha-1(I) procollagen mRNA levels indicating that the potentiation of CDP labeling occurs via a pretranslational mechanism. IGF-I had little effect on the incorporation of [3H]thymidine into bone except in the presence of cortisol. Nevertheless, the ability of cortisol to potentiate the stimulatory effect of IGF-I on CDP labeling was independent of cell replication since the enhancement persisted in the presence of aphidicolin, a DNA synthesis inhibitor. Our findings show that physiological concentrations of cortisol can modulate the responsiveness of cells within cultured fetal rat calvariae to the anabolic effects of exogenous IGF-I.