Granzyme B promotes matrix metalloproteinase-1 (MMP-1) release from gingival fibroblasts in a PAR1- and Erk1/2-dependent manner: A novel role in periodontal inflammation

J Periodontal Res. 2024 Feb;59(1):94-103. doi: 10.1111/jre.13190. Epub 2023 Oct 24.


Objective: To gain insights into how proteases signal to connective tissues cells in the periodontium.

Background: The connective tissue degradation observed in periodontitis is largely due to matrix metalloproteinase (MMP) release by gingival fibroblasts. Granzyme B (GzmB) is a serine protease whose role in periodontitis is undefined.

Methods: Human gingival crevicular fluid (GCF) samples were obtained from sites with periodontal disease and healthy control sites. GzmB was quantified in the GCF ([GzmB]GCF ) by ELISA. Gingival fibroblasts (GF) were cultured in the presence or absence of recombinant GzmB. Culture supernatants were analyzed by ELISA to quantify GzmB-induced release of interstitial collagenase (MMP-1). In some experiments, cells were pre-treated with the inhibitor PD98059 to block MEK/ERK signaling. The protease-activated receptor-1 (PAR-1) was blocked with ATAP-2 neutralizing antibody prior to GzmB stimulation. Systemic MMP-1 levels were measured in plasma from wild-type (WT) and granzyme-B-knockout (GzmB-/- ) mice.

Results: The [GzmB]GCF in human samples was ~4-5 fold higher at sites of periodontal disease (gingivitis/periodontitis) compared to healthy control sites, suggesting an association between GzmB and localized matrix degradation. GzmB induced a ~4-5-fold increase in MMP-1 secretion by cultured fibroblasts. GzmB induced phosphorylation of Erk1/2, which was abrogated by PD98059. GzmB-induced upregulation of MMP-1 secretion was also reduced by PD98059. Blockade of PAR-1 function by ATAP-2 abrogated the increase in MMP-1 secretion by GF. Circulating MMP-1 was similar in WT and GzmB-/- mice, suggesting that GzmB's effects on MMP-1 release are not reflected systemically.

Conclusion: These data point to a novel GzmB-driven signaling pathway in fibroblasts in which MMP-1 secretion is upregulated in a PAR1- and Erk1/2-dependent manner.

Keywords: Granzyme B; MAP kinases; MMP-1; gingival crevicular fluid; periodontitis.

MeSH terms

  • Animals
  • Fibroblasts / metabolism
  • Gingival Crevicular Fluid / chemistry
  • Granzymes
  • Humans
  • Inflammation
  • Matrix Metalloproteinase 1* / metabolism
  • Matrix Metalloproteinase 13 / analysis
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 8 / analysis
  • Mice
  • Periodontitis*
  • Receptor, PAR-1


  • Matrix Metalloproteinase 1
  • Granzymes
  • Receptor, PAR-1
  • Matrix Metalloproteinase 8
  • Matrix Metalloproteinase 13
  • Matrix Metalloproteinase 3