Matrix Metalloproteinase-9 Enhances Osteoclastogenesis: Insights from Transgenic Rabbit Bone Marrow Models and In Vitro Studies

Int J Mol Sci. 2025 Mar 29;26(7):3194. doi: 10.3390/ijms26073194.

Abstract

Osteoclastogenesis is tightly regulated by receptor activator of nuclear factor kappa-B ligand (RANKL) signaling, yet the role of matrix metalloproteinase-9 (MMP-9) in this process remains controversial. We established a high-yield osteoclastogenesis system using cryopreserved rabbit bone marrow cells (1 × 109 cells/femur) treated with Macrophage colony-stimulating factor (M-CSF) and RANKL. Bone marrow cells from MMP-9 transgenic rabbits (macrophage-specific overexpression) and MMP-9-transfected RAW264.7 macrophages were compared to wild-type controls. MMP-9 overexpression increased osteoclastogenesis 5.5-fold (20 ng/mL RANKL, * p < 0.01) while suppressing inflammatory cytokines (IL-1β, TNF-α). RAW264.7 macrophages stably transfected with human MMP-9 similarly exhibited reduced inflammatory cytokine levels and enhanced osteoclastogenesis. MMP-9 acts as a dual regulator of osteoclastogenesis and inflammation, suggesting therapeutic potential for osteoporosis management.

Keywords: MMP-9; bone marrow cells; inflammation; osteoclast; transgenic rabbit.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Bone Marrow Cells* / cytology
  • Bone Marrow Cells* / metabolism
  • Cell Differentiation
  • Cytokines / metabolism
  • Humans
  • Macrophage Colony-Stimulating Factor / pharmacology
  • Macrophages / metabolism
  • Matrix Metalloproteinase 9* / genetics
  • Matrix Metalloproteinase 9* / metabolism
  • Mice
  • Osteoclasts* / cytology
  • Osteoclasts* / metabolism
  • Osteogenesis* / genetics
  • RANK Ligand / metabolism
  • RANK Ligand / pharmacology
  • RAW 264.7 Cells
  • Rabbits

Substances

  • Matrix Metalloproteinase 9
  • RANK Ligand
  • Macrophage Colony-Stimulating Factor
  • Cytokines