Our knowledge of the concentration of growth factors in growing bone is limited. In the present study, we examined the developmental changes in the concentrations of insulin-like growth factor I (IGF-I) and transforming growth factor beta (TGF-β) in the rat femur between weanling and maturity. We show that during the rapid growth phase there is a continuous rise in bone matrix IGF-I and TGF-β in all compartments of the femoral bone. The association between IGF-I and TGF-β is not only temporal, but with few exceptions is also observed within the animals of each age class. These data support the hypothesis that IGF-I and TGF-β play an important role in the growth-associated accumulation of bone mass.

Previous studies have indicated that the mitogenic responsiveness of human bone cells may change with age. In the present study, we examined whether aging affects the capacity of transforming growth factor β (TGF-β) to stimulate the colony formation of human osteoprogenitor cells. Outgrowths of bone cells from 98 iliac crest biopsies were plated at a density of 25 cells/cm2 and cultured for 3 weeks in the presence of 10% fetal calf serum. Approximately 5% of the plated cells gave rise to clonal colonies. TGF-β (10−11 M) significantly increased the estimated number of cells per colony. However, the stimulatory effect of TGF-β significantly declined with donor age (r=−0.26, P=0.01). Whereas TGF-β raised the average number of cells per colony in cultures from donors below the age of 50 years by 136±50%, the average increase was only 43±16% in donors older than 60 years. These data raise the possibility that aging may be associated with a declining capacity of TGF-β to enlarge the pool of bone cells that can be generated from a single human osteoblast progenitor cell.

Insulin-like growth factor-I (IGF-I) plays an important role in bone metabolism, but data on the regulation of IGF-I in bone tissue in vivo are still limited. In the present study, we examined the effects of ovariectomy (ovx) and estrogen replacement on the skeletal concentration of IGF-I in the femur shaft of 6-10 week-old female rats. Ovx had no consistent effect on bone matrix IGF-I concentration regardless of animal age at ovx. In contrast, administration of estradiol in doses that exceeded physiological replacement (50 and 150 nmol/kg per day, subcutaneously) significantly increased the bone matrix IGF-I concentration. These are the first in vivo data which demonstrate that estrogens are capable of increasing the concentration of IGF-I in bone tissue. However, this stimulatory effect appears to be limited to supraphysiological estrogen concentrations.

Transforming growth factor beta (TGF-beta) is thought to play an important role in bone metabolism, but its relationship to human bone turnover and bone mass has not been examined yet. In this study, we measured the concentration of TGF-beta in 811 samples of male and female bone from four representative sites of the human skeleton and in the supernatants of 72 short-term human bone marrow cultures from the iliac crest. The concentrations of TGF-beta1 and TGF-beta2 in the bone matrix were positively correlated with histomorphometric indices of bone resorption and bone formation and with serum levels of osteocalcin and bone-specific alkaline phosphatase. We also observed a positive association between the release of TGF-beta in the bone marrow cultures and serum osteocalcin. Changes in the rate of cancellous or cortical bone remodeling with age or menopause were accompanied by corresponding changes in skeletal TGF-beta. In contrast, there was no significant relationship between the concentration of TGF-beta and bone volume at any skeletal site. In conclusion, our study supports the hypothesis that TGF-beta plays an important role in human bone remodeling, but fails to demonstrate an association between the skeletal concentration of TGF-beta and human bone mass.

Plasminogen activators (PA) and plasminogen activator inhibitors (PAI) have been implicated in the process of extracellular matrix degradation. To study their role in bone matrix turnover, we examined the activity and regulation of PA and PAI in cultures of periosteal osteoblast-like precursor cells and mature osteoblast-like cells from fetal rat calvariae. Both cell populations released PA activity of the tissue type and a 50K PAI species into the culture medium. However, mature osteoblasts had a strikingly lower PA activity and higher PAI activity than periosteal precursor cells, indicating that osteoblast differentiation is associated with a marked decrease in the PA/PAI ratio. PTH and prostaglandin E2 transiently increased PA activity and decreased PAI activity. In contrast, transforming growth factor-beta decreased PA activity and increased PAI activity. Differential effects of these factors on PA and PAI activity may be involved in the regulation of extracellular matrix deposition by osteoblasts.