The early recruitment of inflammatory cells to sites of bone fracture and trauma is a critical determinant in successful fracture healing. Released by infiltrating inflammatory cells, myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are heme-containing enzymes, whose functional involvement in bone repair has mainly been studied in the context of providing a mechanism for oxidative defense against invading microorganisms. We report here novel findings that show peroxidase enzymes have the capacity to stimulate osteoblastic cells to secrete collagen I protein and generate a mineralized extracellular matrix in vitro. Mechanistic studies conducted using cultured osteoblasts show that peroxidase enzymes stimulate collagen biosynthesis at a post-translational level in a prolyl hydroxylase-dependent manner, which does not require ascorbic acid. Our studies demonstrate that osteoblasts rapidly bind and internalize both MPO and EPO, and the catalytic activity of these peroxidase enzymes is essential to support collagen I biosynthesis and subsequent release of collagen by osteoblasts. We show that EPO is capable of regulating osteogenic gene expression and matrix mineralization in culture, suggesting that peroxidase enzymes may play an important role not only in normal bone repair, but also in the progression of pathological states where infiltrating inflammatory cells are known to deposit peroxidases.
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This work was supported in part by The Hospital Research Foundation and the National Health and Medical Research Council (Career Development Fellowship/627015; Project Grant/1050694).
Compliance with Ethical Standards
Conflict of Interest
Mark O. DeNichilo, Alexandra J. Shoubridge, Vasilios Panagopoulos, Vasilios Liapis, Aneta Zysk, Irene Zinonos, Shelley Hay, Gerald J. Atkins, David M. Findlay, and Andreas Evdokiou declare that they have no conflict of interest.
Human and Animal Rights and Informed Consent
The use of all normal human donor-derived bone tissue was approved by the human ethics committees of the Royal Adelaide Hospital/University of Adelaide (Approval No. RAH130114). Human bone samples were obtained with informed written donor consent, as required and approved by the ethics committee.
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