Analysis of gingival epithelial cells and fibroblast using three-dimensional culture system mimic to periodontal pockets
| Project/Area Number |
15592189
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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 |
Periodontal dentistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
NAKAE Hideaki The University of Tokushima, Institute of Health Biosciences of Tokushima Graduate School, Department of Conservative Dentistry, Associate Professor, 大学院・ヘルスバイオサイエンス研究部, 助教授 (30227730)
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| Co-Investigator(Kenkyū-buntansha) |
SHINOHARA Chihiro Tokushima University, Hospital, Department of Oral Care and Clinical Education, Instructor, 医学部・歯学部附属病院, 助手 (50332820)
中西 正 徳島大学, 医学部・歯学部附属病院, 講師 (00217770)
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| Project Period (FY) |
2003 – 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: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | gingival epithelial cell / three-dimensional culture / gingival fibroblast / PIPAAm / tight junction / claudin / tight junction / 上皮細胞 / 線維芽細胞 / 歯周病関連細菌 / 3次元培養 / IL-8 |
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| Research Abstract |
To investigate the mechanisms that periodontopathic bacteria cause periodontal tissue destruction we established the three-dimensional co-culture gingival epithelial cell and fibroblast) system using PIPAAm. Gingival epithelial cells were seeded to Rep Cell plate (CellSeed, Tokyo) and were incubated to confluent. Then, epithelial cells were incubated at 32℃ for 30 min. After the incubation epithelial cells were removed as a cell sheet. Then the epithelial cell sheet was placed onto fibroblasts cultured in type 1 collagen. After co-culture of epithelial cells and fibroblasts for a week, we obtained the three-dimensional gingiva-reconstructive culture mimic to periodontal pockets. We compared the expression of claudin, cadherin, JAM-1, component of tight junction and adherence junction, in normal gingival to that of the three-dimensional gingiva-reconstructive culture. The expression pattern of claudin, cadherin, and JAM-1 in the three-dimensional gingiva-reconstructive culture was identical to that of normal gingival.
Then, we examined the expression pattern of claudins in human periodontal pockets. In human periodontal pockets claudins-1, 4, 7, 12 were expressed constitutively. Moreover, those claudins were detected in three-dimensional gingiva-reconstructive culture. This data suggested that our three-dimensional gingiva-reconstructive culture was a useful tool to investigate the interaction between periodontal pockets and perioidontopathic bacteria. In inflamed tissues, the expression level of claudin-10 was much increased than in normal tissues. Though the mechanism of this phenomenon is unclear, this data suggested that claudin-10 played important role in developing periodontal tissue destruction.
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Report
(3 results)
Research Products
(12 results)
