| Project/Area Number |
13672015
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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 |
Conservative dentistry
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| Research Institution | Aichi Gakuin University School of Dentistry |
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Principal Investigator |
NAKAMURA Hiroshi Aichi Gakuin University School of Dentistry, Department of Endodontics, Professor, 歯学部, 教授 (40064878)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Yoshitaka Aichi Gakuin University School of Dentistry, Department of Endodontics, Assistant professor, 歯学部, 助手 (00329608)
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| Project Period (FY) |
2001 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2001: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Human Periodontal Ligament Cells / Sonicated Bacterial Extracts(SBE) / P.gingivalis / MMP-2 / TIMP / P.sinngivalis |
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| Research Abstract |
Matrix metalloproteinases (MMPs) play a critical role in the homeostasis of the extracellular matrix (ECM). The imbalance between production of MMPs and that of their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), plays an important role in tissue destruction. It has been reported that MMP-1 and MMP-9 are activated by the bacteria associated with the production of periapical diseases. However few reports regarding the activation of pro-MMP-2 and inactivation of TIMP have been published. The purpose of the present study was to evaluate the activation of pro-MMP-2 and inactivation of TIMP in cultured cells.
l )Measurement of MMP-2 activity
The supernatant obtained from medium containing PL cells or HT1080 cells and a sonicated bacterial extract (SEE) from P. gingivalis (P.g.) was analyzed by the gelatin zymography method. When PL cells or HT1080 cells were treated with the SBE, visible bands that corresponded to active MMP-2 were observed on the gelatin zymograms.
2) Measurement of TIMP-1 and TIMP-2 proteins
The amount of TIMP-1 was measured by the EIA method of Kodama et al. TIMP-1 was not detected in the supernatant obtained from the medium of cultures of P.g. SBE-treated PL cells. The amount of TIMP-1 and TIMP-2 in the supernatant obtained from HT1080 cell cultures treated with the extract decreased in a P.g. SBE concentration-dependent manner.
3) Measurement of TIMP-1 activity
After a solution of purified TIMP-1 had been reacted with SBE, TIMP-1 retained in the solution was evaluated by gelatin zymography in terms of its inhibitory activity toward MMP-1 or MMP-2. P.g. SEE remarkably decreased the inhibitory activity of TIMP-1 toward MMP-2.
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