| Project/Area Number |
13672156
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
矯正・小児・社会系歯学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
YOKOYAMA Masaaki The University of Tokushima, School of Dentistry, Research Associate, 歯学部, 助手 (10314882)
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| Co-Investigator(Kenkyū-buntansha) |
TAMATANI Kanako The University of Tokushima, School of Dentistry, Research Associate, 歯学部, 助手 (40243711)
HINODE Daisuke The University of Tokushima, School of Dentistry, Associate Professor, 歯学部, 助教授 (70189801)
NAKAMURA Ryo The University of Tokushima, School of Dentistry, Professor, 歯学部, 教授 (30034169)
NOZOMI Yokoyama The University of Tokushima, School of Dentistry, Research Associate, 歯学部, 助手 (40325270)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2001: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | human gingival epithelial cell / human gingival fibroblast / cytotoxicity assay / MMP-2 / MMP-9 / MMP inhibitor / periodontal pocket formation / keratinocyte migration / ヒト歯肉繊維芽細胞 / 歯周病原細菌 / 歯周ポケット / 歯肉上皮細胞 / 歯肉繊維芽細胞 / MMP / TNFα / TGFβ |
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| Research Abstract |
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.
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