Transforming growth factor-beta and the initiation of chondrogenesis and osteogenesis in the rat femur.

TLDR

The study examined whether exogenous TGF‑β can trigger osteogenesis and chondrogenesis in rat femurs, key processes in bone formation and fracture repair. Subperiosteal injections of TGF‑β1 or TGF‑β2 in newborn rat femurs caused localized intramembranous bone and cartilage formation, with TGF‑β2 generating a 375 % larger mass and dose‑dependent shifts toward bone; after cessation, endochondral ossification replaced cartilage, and TGF‑β2 stimulated autocrine TGF‑β1 production, showing that periosteal mesenchymal precursors proliferate and differentiate in response to TGF‑β, recapitulating embryonic bone formation and fracture healing.

Abstract

We have investigated the ability of exogenous transforming growth factor-beta (TGF-beta) to induce osteogenesis and chondrogenesis, critical events in both bone formation and fracture healing. Daily injections of TGF-beta 1 or 2 into the subperiosteal region of newborn rat femurs resulted in localized intramembranous bone formation and chondrogenesis. After cessation of the injections, endochondral ossification occurred, resulting in replacement of cartilage with bone. Gene expression of type II collagen and immunolocalization of types I and II collagen were detected within the TGF-beta-induced cartilage and bone. Moreover, injection of TGF-beta 2 stimulated synthesis of TGF-beta 1 in chondrocytes and osteoblasts within the newly induced bone and cartilage, suggesting positive autoregulation of TGF-beta. TGF-beta 2 was more active in vivo than TGF-beta 1, stimulating formation of a mass that was on the average 375% larger at a comparable dose (p less than 0.001). With either TGF-beta isoform, the dose of the growth factor determined which type of tissue formed, so that the ratio of cartilage formation to intramembranous bone formation decreased as the dose was lowered. For TGF-beta 1, reducing the daily dose from 200 to 20 ng decreased the cartilage/intramembranous bone formation ratio from 3.57 to zero (p less than 0.001). With TGF-beta 2, the same dose change decreased the ratio from 3.71 to 0.28 (p less than 0.001). These data demonstrate that mesenchymal precursor cells in the periosteum are stimulated by TGF-beta to proliferate and differentiate, as occurs in embryologic bone formation and early fracture healing.