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Abstract
Purpose
To evaluate the temporal and spatial expression of Transforming Growth Factor-β1 and Matrix Metalloproteinase-1 in distraction osteogenesis and fracture healing models.
Materials and Methods
Distraction osteogenesis was performed on the tibial diaphyses of Sprague-Dawley rats (latent period for 1 week, distraction for 2 weeks). The rats were euthanized at each week and the level of mRNA expression was assessed by real-time RT PCR and immunohistochemical staining.
Results
Although the level of TGF-β1 mRNA and MMP-1 mRNA expression was increased during distraction osteogenesis and fracture healing, the level of mRNA expression was significantly higher in the distraction phase in the distraction group than in the fracture healing group at the same phase. After the distraction phase, the level of mRNA expression in both groups decreased to the base line. The peak expression of mRNA was followed by that of TGF-β1 mRNA. Immunohistochemical staining revealed that TGF-β1 was expressed mainly in the osteoblast and endothelial cells, and MMP-1 was expressed mainly in the endothelial cells of the vessel.
Figures and Tables
Fig. 1
Application of distraction osteogenesis. (A) Appearance of external fixator on a mouse. (B) Medial longitudinal incision and corticotomy.
Fig. 2
Serial examination of H&E and immunohistochemical staining in distraction osteogenesis. Seven days after the osteotomy, Hematoma organization and angiogenesis were observed. The expression of TGF-β1 and MMP-1 in the endothelial and proliferating mesenchymal cells. At 14 days, a zonal phenomenon was noted. MCF, Microcolumn formation; PMF, Primary mineralization front; FIZ, Fibrous interzone. Expression of TGF-β1 in osteoblasts in PMF was observed. At 21 days, angiogenesis was also observed in the FIZ. At 28 days, there was an increase in the amount of woven bone and extracellular matrix.
Fig. 3
Serial examination of H&E and immunohistochemical staining in the fracture healing model. At 14 days, enchondral ossification was observed. TGF-β1 expression was observed in the small proliferative chondrocytes, vessel wall and osteoblasts but not in the hypertropic chondrocytes. MMP-1 expression was observed only in the vessel wall. At 21 days, TGF-β1 expression was noted in the osteoblasts around the woven bone and vessel wall. At 28 days, the fracture gap was connected with new bone. TGF-β1 was expressed weakly on the vessel wall, and MMP-1 was expressed in the vessel wall.
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