Increased Lengthening Rate Decreases Expression of Fibroblast Growth Factor 2, Platelet-Derived Growth Factor, Vascular Endothelial Growth Factor, and CD31 in a Rat Model of Distraction Osteogenesis

Abstract

The rate of lengthening has a profound impact on bone regeneration during distraction osteogenesis. Rapid distraction can delay or completely inhibit union, whereas distracting too slowly may lead to premature consolidation. However, the mechanisms responsible for retardation of healing due to rapid distraction have not been elucidated. This study explored whether rapid distraction alters the expression of certain angiogenic growth factors, in particular, fibroblast growth factor 2 (FGF-2), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF-AA), and subsequent new vessel formation as evidenced by platelet endothelial cellular adhesion marker expression (CD31), an indicator of vascular budding. Unilateral femoral lengthenings were performed in 60 male Sprague-Dawley rats using a protocol that involved a 7-day latency period and distraction rates of either 0.5 (slow distraction) or 1.5 mm/d (fast distraction) for a total of 7.0 mm of lengthening. Animals were euthanized on postoperative days 8, 10, 12, 14, and 21 (n = 6 per time point and distraction rate). Expression of FGF-2, VEGF, PDGF-AA, and CD31 was characterized immunohistochemically. Cellular staining of FGF-2, PDGF-AA, VEGF, and CD31 was reduced on days 8 to 12 in the regenerate of the fast-distraction animals compared with the slow-distraction animals. Staining of all growth factors was weak on days 14 and 21 at the slow rate and absent at the fast rate. Regardless of time point, a similar spatial localization of growth factor expression was observed at the 2 rates of distraction. The reduced expression of angiogenic growth factors and CD31, a marker of new vessel formation, indicates that the angiogenic cascade and new vessel formation required for effective bone healing is disrupted at a distraction rate of 1.5 mm/d in a rat model of limb lengthening. Delayed bone healing with rapid distraction may be due in part to decreased cellular signaling required for angiogenesis. It may be possible to improve bone healing at increased distraction rates with the appropriately timed administration of growth factors.