| Project/Area Number |
10470446
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | OKAYAMA UNIVERSITY |
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Principal Investigator |
YAMAMOTO Teruko Okayama University Dental School, Professor, 歯学部, 教授 (00127250)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Kazuo Okayama University Dental School, Research assistant, 歯学部, 助手 (50304316)
MIYAMOTO Manabu Okayama University Dental School, Research assistant, 歯学部, 助手 (40252978)
YAMASHIRO Tkashi Okayama University Dental School, Associate Professor, 歯学部, 講師 (70294428)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥9,800,000 (Direct Cost: ¥9,800,000)
Fiscal Year 1999: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1998: ¥6,500,000 (Direct Cost: ¥6,500,000)
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| Keywords | periodontal ligament / Clonal analysis / Alklinephosphatase / trkA / in situ ハイブリダイゼーション |
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| Research Abstract |
Clonal analysis of periodontal ligament (PDL) cells isolated from human extracted teeth was performed for establishing a culture method of homologous, single cell derived PDL cells, and for elucidating roles of PDL cells in orthodontic tooth movement.
Human PDL was excised from a premolar or a third molar, which were extracted from orthodontic reasons, minced and placed onto culture dish with Dalbecco's modified eagle medium (DMEM) containing 10 % fetal bovine serum. After 10 to 14 days fibroblastic cells were extended and then round-shaped cells were extended from PDL tissue. After 20 days round-shaped cells surrounded PDL tissue completely and fibroblastic cells were extended outside of round-shaped cells. Other shaped cells were also observed in microscopy. All cells lost their own potentials within 15 to 20 subcultures in morphologically and in cellular growth. Next experiments were done for establishing a novel isolation method for clone from a single-cell derived PDL cell. Treatme
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nts with collagenase and trypsin to fresh human PDL tissue were successfully isolated a single PDL cells with minimum cell damage. Although a majority of cells isolated by above method was fibroblastic cells, different shaped cells were isolated. Some cells were stained immunologically for cytokeratin antibody. These cells were thought to be from Malassez epithelial rest. We also observed that trkA, a neurotrophin receptor, was expressed at epithelial cells in Malassez epithelial rest. Then the study for trkA expression in primary PDL culture is in progress. We are also in progress for analyzing immunological and molecular properties of single-cell derived cells. Furthermore, several clones form single-cell derived cells were transplanted subcutaneously into nude mice for evaluating their differentiation capacities in vivo. We have also investigated the roles for PDL cells during tooth movement in rat. The reports of these observations are submitted to the international journals, and some of them have been accepted. Less
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