Increased matrix metalloproteinase-2 in ligamentum flavum hypertrophy and the regulation of MMP-2/TIMPs by elastin-derived peptides

J Orthop Res. 2024 Sep;42(9):2061-2071. doi: 10.1002/jor.25841. Epub 2024 Mar 28.

Abstract

The study aimed to examine matrix metalloproteinase-2 (MMP-2) expression in a rat ligamentum flavum (LF) hypertrophy model in vivo, and the effect of elastin-derived peptides (EDPs) on MMP-2 and tissue inhibitors of metalloproteinases (TIMPs) in rat LF cells in vitro. Surgical destabilization was performed at the rat spinal L3/4 level to induce increased mechanical stress. Rats were killed at 6- and 12-weeks postsurgery for histological staining, immunohistochemical staining, RT-qPCR and western blot. 100 µg/mL EDPs were applied to isolated normal rat LF cells, with or without pretreatment of elastin receptor complex (ERC) inhibitors, to assess the expression of MMP-2, TIMP-1, and TIMP-2. Spinal destabilization led to LF hypertrophy, observed through increased LF thickness and area, along with histological changes of chondrometaplasia and elastic fiber degradation. LF was also stained positively for Col I and Col II, where elastic fiber has broken down. MMP-2 expression was notably elevated in the hypertrophied LF, accompanied by increased TIMP-2 and TIMP-3 levels. EDPs were found to suppress MMP-2 expression and reduce TIMP-1 and TIMP-2 levels in rat LF cells. Interestingly, exposure to EDPs led to a significant rise in MMP-2/TIMP-1 and MMP-2/TIMP-2 ratios, dependent on the ERC. Collectively, the study suggests that increased MMP-2 activity contributes to elastic fiber degradation in hypertrophied LF, generating EDPs that further enhance the MMP-2/TIMPs ratio in LF cells in an ERC-dependent manner. Further research is essential to delve into the mechanisms of EDPs in LF hypertrophy.

Keywords: MMP‐2; TIMPs; elastin‐derived peptides; ligamentum flavum; rat model.

MeSH terms

  • Animals
  • Elastin* / metabolism
  • Hypertrophy*
  • Ligamentum Flavum* / metabolism
  • Ligamentum Flavum* / pathology
  • Male
  • Matrix Metalloproteinase 2* / metabolism
  • Peptides / metabolism
  • Peptides / pharmacology
  • Rats
  • Rats, Sprague-Dawley*
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism
  • Tissue Inhibitor of Metalloproteinase-2* / metabolism

Substances

  • Matrix Metalloproteinase 2
  • Elastin
  • Tissue Inhibitor of Metalloproteinase-2
  • Mmp2 protein, rat
  • Tissue Inhibitor of Metalloproteinase-1
  • Timp2 protein, rat
  • TIMP1 protein, rat
  • Peptides