Study on the mechanism of periodontal pocket formation using Matrix Metalloproteinase inhibitors

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The aim of this study was to clarify the behavior of Matrix Metalloproteinases (MMPs) produced by gingival cells in relation to the mechanism of periodontal pocket formation.Cultured human gingival fibroblast (HGF), cultured human gingival epithelial cell (HE), and their culture supernatants were used for all experiments. By gelatin zymography, MMP-9 from HE and MMP-2 from HGF were detected clearly, while MMP-2 from HE were faint and MMP-9 from HGF ware detected only when HGF were stimulated by TNFα. Three kinds of hydroxamic acid based MMP inhibitors (ONO-4817, ONO-MI1-514, ONO-MI1-570) were tested on these MMP activity. First, cytotoxic assay was performed to verify the safety toward host cells. The cell viability and the cell growth of HGF and HE were not suppressed by all inhibitors at final concentration of 50μM. By gelatin zymography, it was found that all inhibitors clearly inhibited MMP-2 and MMP-9 produced by HGF or HE, and the intensity of their inhibitory effect was found in order of ONO-4817, ONO-MI1-514, ONO-MI1-570. The same effect was clarified with an experiment using a colorimetric assay kit. Further investigation revealed that all inhibitors acted in a dose dependent manner. To clarify the role of MMP derived from gingival cells during a periodontal pocket formation, we investigated the effects of MMP inhibitors on keratinocyte migration in vitro.These data suggest that MMP, especially MMP-9 may participate in a pocket formation, which is concomitant with epithelial cell migration, and also support the possibility that these MMP inhibitors could be applied to the patients with periodontal disease as preventive and/or therapeutic use.