| Project/Area Number |
14207080
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | Osaka University |
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Principal Investigator |
EBISU Shigeyuki Osaka University, Graduate School of Dentistry, Professor, 大学院・歯学研究科, 教授 (50116000)
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| Co-Investigator(Kenkyū-buntansha) |
NOIRI Yuichiro Osaka University, Graduate School of Dentistry, Instructor, 大学院・歯学研究科, 助手 (50218286)
成松 雅博 大阪大学, 大学院・歯学研究科, 助手 (80362680)
金森 市朗 大阪大学, 大学院・歯学研究科, 日本学術振興会特別研究員
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥43,030,000 (Direct Cost: ¥33,100,000、Indirect Cost: ¥9,930,000)
Fiscal Year 2004: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2003: ¥10,530,000 (Direct Cost: ¥8,100,000、Indirect Cost: ¥2,430,000)
Fiscal Year 2002: ¥23,270,000 (Direct Cost: ¥17,900,000、Indirect Cost: ¥5,370,000)
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| Keywords | Refractory periapical periodontal disease / Extraradicular biofilm / Gene analysis / Porphyromonas gingivalis / modified Robbins Device / Antimicrobial agent / Gutta-percha point / Er:YAG laser / 細菌バイオフィルム / 免疫組織化学 / 局在性 / Er : YAGレーザー / 微細形態学 / 超微形態学 / Modified Robbins Device / 16S rRNA / クローニング / 自己凝集 |
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| Research Abstract |
We investigated the identification and localization of the bacteria in human extraradicular bolls. Extraradicular biofilms to identify bacteria using a PCR-based 16S rRNA gene assay, and to observe immunohistochemical localization of selected bacterial species were taken from patients with refractory periapical periodontitis. Fusobacterium nucleatum, Porphyromonas gingivalis, and Tannellera forsythensis were frequently detected. P.gingivalis were detected in all parts of the extraradicular biofilms. F.nucleatum were predominantly scattered at the middle layer, and T.forsythensis located at the superfacial layer in the biofilms.
On the other hand, the control methods against single species biofilms of bacteria identified from the extraradicular biofilm were examined. P.gingivalis strain 381 biofilms were prepared using modified Robbins Device. Minocycline hydrochloride and Metronidazole, which showed effect against planktonic P.gingivalis cells, were not effective though chlorhexidine gl
… More
uconate were effective. Moreover, we found that the effects of Er.YAG laser against Enterococcus faecalis, Streptococcus mutans, Actinomyces viscosus and Propionibacterium acnes biofilms were different.
We analyzed the gene associated with biofilm formation of P.gingivalis. A putative glycosyltransferase gene was isolated from P.gingivalis strain 381 and designated as gtfA. GtfA-deficient P.gingivalis lacked mature fimbriae morphologically and showed quite a low ability for autoaggregation, and its ability for attachment to epithelial cells was severely impaired. These results suggested that gtfA might regulate attachment ability in subgingival biofilms and act has a potent virulence factor.
The initial biofilm-forming ability of root canal isolates on gutta-percha points was investigated. Enterococcus faecalis, Streptococcus sanguis, Streptococcus intermedius, Streptococcus pyogenes and Staphyrococcus aureus biofilms were generated on the surfaces of the specimens incubated in culture mediums supplemented with 45 or 90 v% serum. E.faecalis and S.sanguis biofilms were significantly thicker than those of S.intermedius, S.pyogenes and S.aureus. No biofilms were detected on the specimens incubated with F.nucleatum, P.acnes, P.gingivalis, and Prevotella intermedia. Less
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