TGF-β1 As Possible Link between Loss of Bone Mineral Density and Chronic Inflammation

Background: The TGF family plays a key role in bone homeostasis. Systemic or topic application of proteins of this family apparently positively affects bone healing in vivo. However, patients with chronic inflammation, having increased TGF-beta(1) serum-levels, often show reduced bone mineral content and disturbed bone healing. Therefore, we wanted to identify intracellular mechanisms induced by chronic presence of TGF-beta(1) and their possible role in bone homeostasis in primary human osteoblasts. Methodology/Principal Findings: Osteoblasts were isolated from femur heads of patients undergoing total hip replacement. Adenoviral reporter assays showed that in primary human osteoblasts TGF-beta(1) mediates its signal via Smad2/3 and not Smad1/5/8. It induces proliferation as an intermediate response but decreases AP-activity and inorganic matrix production as a late response. In addition, expression levels of osteoblastic markers were strongly regulated (AP down arrow; Osteocalcin down arrow; Osteopontin up arrow; MGP down arrow; BMP 2 down arrow; BSP2 down arrow; OSF2 down arrow; Osteoprotegerin down arrow; RANKL up arrow) towards an osteoclast recruiting phenotype. All effects were blocked by inhibition of Smad2/3 signaling with the Alk5-Inhibitor (SB431542). Interestingly, a rescue experiment showed that reduced AP-activities did not recover to base line levels, even 8 days after stopping the TGF-beta(1) application. Conclusions/Significance: In spite of the initial positive effects on cell proliferation, it is questionable if continuous Smad2/3 phosphorylation is beneficial for bone healing, because decreased AP-activity and BMP2 levels indicate a loss of function of the osteoblasts. Thus, inhibition of Smad2/3 phosphorylation might positively influence functional activity of osteoblasts in patients with chronically elevated TGF-beta(1) levels and thus, could lead to an improved bone healing in vivo.