THYROID-HORMONE SUPPRESSES THE DIFFERENTIATION OF OSTEOPROGENITOR CELLS TO OSTEOBLASTS, BUT ENHANCES FUNCTIONAL ACTIVITIES OF MATURE OSTEOBLASTS IN CULTURED RAT CALVARIA CELLS

The effects of thyroid hormone on osteoblastic differentiation and act ivity were studied in fetal rat calvaria (RC) cells cultured for up to 30 days in medium supplemented with thyroid hormone-depleted serum. I n this condition, the cells proliferated and differentiated to form mi neralized bone nodules (BN) and expressed osteoblastic markers such as alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). The continuous presence of triiodothyronine (T3) at 10(-9)-10(-8) M i n the medium inhibited the osteoblastic differentiation: 34% decrease in ALP activity on day 12 and 60% decrease in BN formation on day 15 a t 10(-8) M. T3 at these doses had no effect on the DNA content of RC c ells at confluence (day 6). Short-term (48-h) exposure of T3 at 10(-9) M or higher decreased ALP activity when RC cells were differentiating (days 7-11). However, when BN formation by the cells had already reac hed a plateau (day 28), the activity was increased by treatment with T 3 at 10(-7)-10(-6) M. OCN production was increased dose dependently by this treatment with T3 (2.1-fold and 1.3-fold of control at 10(-8) M on days 11 and 28, respectively). Similar increases were observed in t he levels of OCN mRNA. In addition, increases in phosphorylated OPN in the medium (day 11) and mineralized matrix (day 28) were observed (1. 5-fold at 10(-8)-10(-6) M), while OPN synthesis and the level of its m RNA were depressed by T3 (60-70% of control at 10(-8) M). These result s suggest that T3 regulates osteoblastic differentiation and activity depending on the state of cell differentiation: T3 suppresses the diff erentiation of osteoprogenitor cells to osteoblasts, but enhances the functional activity of mature osteoblasts. (C) 1994 Wiley-Liss, Inc.