Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
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NOX4 and its association with myeloperoxidase and osteopontin in regulating endochondral ossification

  • Kayoung Ko (BK21 Four Project, Department of Medical Sciences, Soonchunhyang University) ;
  • Seohee Choi (BK21 Four Project, Department of Medical Sciences, Soonchunhyang University) ;
  • Miri Jo (BK21 Four Project, Department of Medical Sciences, Soonchunhyang University) ;
  • Chaeyoung Kim (BK21 Four Project, Department of Medical Sciences, Soonchunhyang University) ;
  • Napissara Boonpraman (Department of Translational Neuroscience, College of Human Medicine, Michigan State University) ;
  • Jihyun Youm (Department of Gerontology, Graduate School of East-West Medical Science, Kyunghee University) ;
  • Sun Shin Yi (BK21 Four Project, Department of Medical Sciences, Soonchunhyang University)
  • Received : 2024.02.26
  • Accepted : 2024.05.13
  • Published : 2024.07.31
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Abstract

Importance: Endochondral ossification plays an important role in skeletal development. Recent studies have suggested a link between increased intracellular reactive oxygen species (ROS) and skeletal disorders. Moreover, previous studies have revealed that increasing the levels of myeloperoxidase (MPO) and osteopontin (OPN) while inhibiting NADPH oxidase 4 (NOX4) can enhance bone growth. This investigation provides further evidence by showing a direct link between NOX4 and MPO, OPN in bone function. Objective: This study investigates NOX4, an enzyme producing hydrogen peroxide, in endochondral ossification and bone remodeling. NOX4's role in osteoblast formation and osteogenic signaling pathways is explored. Methods: Using NOX4-deficient (NOX4-/-) and ovariectomized (OVX) mice, we identify NOX4's potential mediators in bone maturation. Results: NOX4-/- mice displayed significant differences in bone mass and structure. Compared to the normal Control and OVX groups. Hematoxylin and eosin staining showed NOX4-/- mice had the highest trabecular bone volume, while OVX had the lowest. Proteomic analysis revealed significantly elevated MPO and OPN levels in bone marrow-derived cells in NOX4-/- mice. Immunohistochemistry confirmed increased MPO, OPN, and collagen II (COLII) near the epiphyseal plate. Collagen and chondrogenesis analysis supported enhanced bone development in NOX4-/- mice. Conclusions and Relevance: Our results emphasize NOX4's significance in bone morphology, mesenchymal stem cell proteomics, immunohistochemistry, collagen levels, and chondrogenesis. NOX4 deficiency enhances bone development and endochondral ossification, potentially through increased MPO, OPN, and COLII expression. These findings suggest therapeutic implications for skeletal disorders.

Keywords

Acknowledgement

This research was supported by the National Research Foundation (NRF) NRF-2018R1D1A3B07047960, and Soonchunhyang University Research Fund.

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