| Project/Area Number |
08835022
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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 |
咀嚼
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| Research Institution | KANAGAWA DENTAL COLLEGE |
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Principal Investigator |
KAWASE Toshio Kanagawa Dental College, School of Dentistry, Professor, 歯学部, 教授 (30084784)
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| Co-Investigator(Kenkyū-buntansha) |
NOZAKI Naohito Kanagawa Dental College, School of Dentistry, Research Associate, 歯学部, 助手 (70222198)
NISHIYAMA Katsuhiro Kanagawa Dental College, School of Dentistry, Instructor, 歯学部, 講師 (20084783)
SAITO Shigeru Kanagawa Dental College, School of Dentistry, Professor, 歯学部, 教授 (80084713)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Human Periodontal Ligament Fibroblasts / Mechanical Stress / Osteonectin / Tissue Regeneration / Cell Attachment and Spreading Factors / 細胞接着因子 / 細胞外基質 / オステオネクチン / ヒト歯根膜線維芽細胞 / 歯根膜由来線維芽細胞(HPLF) / 歯槽骨由来骨芽細胞(HABC) / Flexcer Strain Unit |
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| Research Abstract |
Periodontal ligament lies between tooth and alveolar bone and functions as a cushion mitigating mechanical stress such as occlusion and mastication, and the mechanical stress may affect the periodontal metabolism. The object of this study was to demonstrate the effect the mechanical stretching on the level of ^<35>S-Met labeled proteins and the synthesis and the gene expression of osteonectin (ON) in human periodontal ligament-derived fibroblasts (HPLF). HPLF were cultured on Flex I,type I collagen coated flexible-bottomed culture plates on a Flexercell strain unit. HPLF were flexed at 3 cycles (10 sec strain and 10 sec relaxation) at 3 (7,18 and 24% increase in surface area) for 1,3 and 5 days).
After macromolecules in media labeled with ^<35>S-Met applied to IEC-DEAE column eluted with 0-0.5M NaCl in PBS (-), SDS-PAGE (5-15%gel) were performed and autoradiograms were obtained. The application of stretching increased of several radiolabeled bands between 200 to 100 and 70 and 30 kDa compared with the non-stress control were shifted to the neutral fraction of of ion-exchange chromatography. The shift of elution position suggested that there may be two possibilities which are the decrease of posttranslational modification such as phosphorylation and acidic glycosylation or enzymatic degradation of modifications.
We have focused to ON which has established as one of bone non-collagenous proteins regulating bone mineralization and remodeling. The synthesis of ON subjected to mechanical stress, as examined by Western blotting analysis were increased in HPLF by 18% for the first day, but results were not significantly different as compared to the synthesis of ON to mechanical stress of 7 and 24%. These findings suggest that ON may be involved in periodontal metabolism in response to the mechanical stress.
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