Experimental Strain-Dependent Control of Transforming Growth Factor- 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 suchasmatrixsynthesis, therateofcell replication, and enzyme activities. The authors further asked whether these changes relate to differences in the biochemical response of osteoblasts totransforminggrowthfactor(TGF), 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 forceassociatedwithadistraction 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 TGFat 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 TGFon this event. However, neither intermittent nor continuous strain significantly altered collagen or noncollagen protein synthesis or the relative effect of TGFon 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 TGFtreatment, and even more so in response to both conditions. In addition, TGFbinding to the type III TGFreceptor was increased in proportion to strain intensity. Conclusions: This study shows that cyclic strain can alter osteoblast activity in multiple ways and predicts that TGFhas different effects during thedistractionprocessonosteoblastsandtherefore 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. (Plast. Reconstr. Surg. 116: 224, 2005.)