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Heparanase mRNA expression during fracture repair in mice

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Abstract

Bone fracture healing takes place through endochondral ossification where cartilaginous callus is replaced by bony callus. Vascular endothelial growth factor (VEGF) is a requisite for endochondral ossification, where blood vessel invasion of cartilaginous callus is crucial. Heparanase is an endoglucuronidase that degrades heparan sulfate proteoglycans (HSPG) and releases heparin-binding growth factors including VEGF as an active form. To investigate the role of heparanase in VEGF recruitment during fracture healing, the expression of heparanase mRNA and VEGF, and vessel formation were examined in mouse fractured bone. On days 5 and 7 after the fracture, when mesenchymal cells proliferated and differentiated into chondrocytes, heparanase mRNA was detected in osteo(chondro)clasts and their precursors, but not in the inflammatory phase (day 3). On day 10, both VEGF and HSPG were produced by hypertrophic chondrocytes of the cartilaginous callus and by osteoblasts of the bony callus; numerous osteo(chondro)clasts resorbing the cartilage expressed strong heparanase signals. Adjacent to the cartilage resorption sites, angiogenesis with CD31-positive endothelial cells and osteogenesis with osteonectin-positive osteoblasts were observed. On days 14 and 21, osteoclasts in the woven bone tissue expressed heparanase mRNA. These data suggest that by producing heparanase osteo(chondro)clasts contribute to the recruitment of the active form of VEGF. Thus osteo(chondro)clasts may promote local angiogenesis as well as callus resorption in endochondral ossification during fracture healing.

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Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan, to S.K. and R.K.. The authors thank Atsumi Kodan, Shuichi Matsuda, and Aiko Kaide for their excellent technical assistance.

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Authors and Affiliations

  1. Division of Molecular Pathology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, 650-0017, Kobe, Japan

    Masamichi Saijo, Riko Kitazawa, Sakan Maeda & Sohei Kitazawa

  2. Discovery Research, Tsukuba Research Institute, Novartis Pharma K. K., Okubo 8, Tsukuba, 300-2611, Ibaraki, Japan

    Motowo Nakajima

  3. Division of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, 650-0017, Kobe, Japan

    Masahiro Kurosaka

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  1. Masamichi Saijo
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  2. Riko Kitazawa
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  3. Motowo Nakajima
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  4. Masahiro Kurosaka
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  5. Sakan Maeda
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  6. Sohei Kitazawa
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Correspondence to Sohei Kitazawa.

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Saijo, M., Kitazawa, R., Nakajima, M. et al. Heparanase mRNA expression during fracture repair in mice. Histochem Cell Biol 120, 493–503 (2003). https://doi.org/10.1007/s00418-003-0589-1

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  • DOI: https://doi.org/10.1007/s00418-003-0589-1

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