Strain-dependent control of transforming growth factor-beta function in osteoblasts in an in vitro model: biochemical events associated with distraction osteogenesis.
BACKGROUND
Distraction osteogenesis is an important clinical method for increasing bone mass, but its effects on bone-forming cells are not well understood. In this study, the authors asked how the mechanical forces that occur during this procedure alter specific osteoblast activities such as matrix synthesis, the rate of cell replication, and enzyme activities. The authors further asked whether these changes relate to differences in the biochemical response of osteoblasts to transforming growth factor-beta (TGF-beta), a potent regulator of bone formation.


METHODS
Osteoblasts were plated on flexible, collagen-coated membranes. One group was unstrained, a second group experienced a single maximum strain load once every 6 hours to simulate intermittent force associated with a distraction protocol of four screw turns per day, and a third group was strained continuously for 24 hours. In the third group, some cell cultures were allowed to recover from strain before analysis. Subsequently, each group was treated with vehicle or TGF-beta at 12 pM (0.3 ng/ml) or 120 pM (3 ng/ml). Data were collected from a minimum of 15 replicate cell culture wells obtained from at least three separate primary culture preparations. Results were assessed with statistical software. Differences were considered significant with values of p < 0.05.


RESULTS
Both strain protocols increased basal osteoblast DNA synthesis but suppressed the relative stimulatory effect of TGF-beta on this event. However, neither intermittent nor continuous strain significantly altered collagen or noncollagen protein synthesis or the relative effect of TGF-beta on these processes in osteoblasts. Basal alkaline phosphatase activity, an intermediate marker of osteoblast differentiation and an early marker of matrix mineralization, decreased significantly in response to continuous strain or to TGF-beta treatment, and even more so in response to both conditions. In addition, TGF-beta binding to the type III TGF-beta receptor was increased in proportion to strain intensity.


CONCLUSIONS
This study shows that cyclic strain can alter osteoblast activity in multiple ways and predicts that TGF-beta has different effects during the distraction process on osteoblasts and therefore on their ability to effect bone formation. They further indicate that mechanical load permits early aspects of osteoblast activation but delays in part later biochemical parameters associated with mineralization to allow new bone growth before consolidation.
Knoll
Bianka I
BI
Department of Surgery, Yale University School of Medicine, New Haven, Conn, USA.
McCarthy
Thomas L
TL
Centrella
Michael
M
Shin
Joseph
J
D000818:Animals; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D006133:Growth Substances / Q000502:physiology*; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology*; D019857:Osteogenesis, Distraction; D011506:Proteins / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013314:Stress, Mechanical; D015854:Up-Regulation
D016326:Extracellular Matrix Proteins; D006133:Growth Substances; D011506:Proteins