Open Access, Peer-reviewed
pISSN 1226-2102
eISSN 2005-8918
    J Korean Orthop Assoc. 2008 Oct;43(5):529-538. Korean.
    Published online Oct 31, 2008.
    https://doi.org/10.4055/jkoa.2008.43.5.529
    Copyright © 2008 The Korean Orthopaedic Association
    Original Article

    Matrix Metalloproteinase-1 and Transforming Growth Factor-β1 Expression during Distraction Osteogenesis and Fracture Healing of the Rat

    Soo-Yong Kang, M.D., Ho-Joong Jung, M.D., Young-Bok Jung, M.D., Eui Chan Jang, M.D., Eun-Yong Lee, M.D., Mi-Kyung Lee, M.D.,* and Mi-Kyung Kim, M.D.†
      • Department of Orthopaedics, College of Medical, Chung-Ang University, Seoul, Korea.
      • *Department of Laboratory Medicine, College of Medical, Chung-Ang University, Seoul, Korea.
      • †Department of Pathology, College of Medical, Chung-Ang University, Seoul, Korea.
    • Address reprint requests to Ho-Joong Jung, M.D. Department of Orthopaedic Surgery, Medical Center of Chung-Ang University, 224-1, Heuksuk-dong, Dongjak-gu, Seoul 156-755, Korea. Tel: +82.2-6299-1588, Fax: +82.2-822-1710, Email: sunu@cau.ac.kr

    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.

    Conclusion

    There is specific time sequence in the expression of TGF-β1 and MMP-1 during fracture healing and distraction osteogenesis. These results suggest that TGF-β1 expression might be associated with the angiogenesis induced by MMP-1 expression during new bone formation.

    Keywords
    Distraction osteogenesis; Fracture healing; TGF-β1; MMP-1; Real-time RT PCR

    Figures

    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.

    Fig. 4
    mRNA expression analysis by real-time RT PCR across the time course of distraction osteogenesis and fracture healing. * Indicate significances compared with the fracture healing group (*p<0.05). DO, Distraction osteogenesis; FH, Fracture healing. (A) TGF-β1 mRNA, (B) MMP-1 mRNA.

    Tables

    Table 1
    List of PCR Primers and the Product Size

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    MeSH Terms
    Animals
    Diaphyses
    Endothelial Cells
    Fracture Healing
    Matrix Metalloproteinase 1
    Osteoblasts
    Osteogenesis, Distraction
    Polymerase Chain Reaction
    RNA, Messenger
    Rats
    Transforming Growth Factor beta1
    Metrics
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