| Project/Area Number |
08457521
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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 |
補綴理工系歯学
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
AKAO Masaru Tokyo Medical and Dental University, Institute for Medical and Dental Engineering, Associate Professor, 医用器材研究所, 助教授 (50143607)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAGI Tohru Tokyo Medical and Dental University, Faculty of Dentistry, Lecturer, 歯学部, 講師 (20124696)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | biomaterials / hydroxyapatite / TCP / cell attachment / osteoblast / focal contact / vinculin / 細胞接着 |
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| Research Abstract |
In the present study, attachment of cultured osteoblasts on the surfaces of biomaterials was Investigated by immunofluorescence microscopy. Osteoblastic UMR 106 cells were cultured on discs (10 mm diameter and 2 mm thickness) of dense hydroxyapatite (HA), alpha-TCP, beta-TCP, HA-coated titanium, Ti-6A1-4V alloy, ultra-high molecular weight polyethylene (UHMWPE) and calcium phosphate/copoly-L-lactide composite.
After 24 h incubation, formation of focal contacts on these materials was examined by indirect immunofluorescence microscopy, followed by staining with monoclonal anti-vinculin. HA-coated titanium and calcium phosphate/copoly-L-lactide composite were not suitable for immunofluorescence observation due to the surface roughness and auto-fluorescence. Cells grown on the surfaces of HA, alpha-TCP, beta-TCP and Ti-6A1-4V were well spread and formed vinculin-containing focal contacts. There was no significant difference in the focal adhesions on these four materials. In contrast, cells grown on UHMWPE were more rounded and less spread. As a results of image analysis, HA has the greatest cell areas and UHMWPE, the smallest ones.
UMR 106 cells were stained with above-mentioned method using monoclonal anti-talin and monoclonal anti-integrins. No significant immunofluorescent labelling of talin and integrins was obtained in the present study. This may be due to the species-specific differences in talin and integrin. HOS cells derived from human osteosarcoma were stained with the same method using monoclonal anti-vinculin. HOS cells were well spread on the surfaces of four materials and formed focal contacts. Cell areas of HOS cells were greater than those of UMR 106 cells. This result indicated that HOS cells produced large-mounts of fibronectin as an extracellular matrix protein in vitro.
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