| Project/Area Number |
14571991
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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 | Kanagawa Dental College |
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Principal Investigator |
DEGUCHI Shinji Kanagawa Dental College, Dentistry, Professor, 歯学部, 教授 (60121018)
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| Co-Investigator(Kenkyū-buntansha) |
SUGAYA Akira Kanagawa Dental College, Dentistry, Associate Professor, 歯学部, 助教授 (30162853)
IKAI Hiroyuki Kanagawa Dental College, Dentistry, Lecturer, 歯学部, 講師 (80277904)
KOKE Yukiko Kanagawa Dental College, Dentistry, Assistant, 歯学部, 助手 (10339781)
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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 |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | periodontum derived cells / regeneration / spheroid / cell growth |
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| Research Abstract |
The ultimate goal of periodontal therapy is the regeneration of destroyed periodontal tissues, and is in the original form and recovery of a function. It is thought that the re-adhesion by the new bone, new cementum, and new connective tissue is required for the destroyed rebirth of an organization. We previously reported that the spheroid was produced with periodontal ligament fibroblasts and might be used for periodontal reconstructive therapy. Because spheroid has spherical 3-dimensional structure and that contained the cells which have cell proliferation activity, and produced extracellular matrix proteins.
This time, we produced spheroid using the human alveolar bone derived cell(HABC) with containing fetal calf serum medium(D-MEM). We performed an immunohistochemical study for spheroids using the monoclonal antibody against collagen type-I, osteopontin(OP), osteocalcin(OC) and Ki-67 antibodies. Another approach is to establish of the culture method with growth factor which were a recombinant human platelet derived growth factor(rhPDGF) and a recombinant human b-fibroblast growth factor(rhb-FGF), in the serum free medium to introducing the periodontal tissue regeneration therapy with the spheroids.
The positive reaction of Ki-67 was observed in the whole of spheroid on the first day and the third day, but observed only in the surface layer on the seventh day and the 21^<st> day. Collagen type-I was observed on the fist day to on the 21^<st> day and an alkaline phosphatase was observed in the whole spheroid on the first day to 7^<th> day. HABC could produce OC and OP at inside surface of the spheroid on the 21^<st> day. The culture with serum free medium contained with 100 ng/ml rhb-FGF appeared the highest growth rate was 42.0 % of the culture with D-MEM at the 5 day. HABC expressed Collagen type-I, OC and OP in the spheroid environment, so spheroids may be capable of being applied to periodontal reconstructive therapy.
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