| Project/Area Number |
01870080
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B).
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| Allocation Type | Single-year Grants |
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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 |
SATO Atsushige Tokyo Medical and Dental University, Faculty of Dentistry, Department of Biomaterials Science, Professor, 歯学部, 教授 (40045985)
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| Co-Investigator(Kenkyū-buntansha) |
HONGO Toshio Tokyo Medical and Dental University, Faculty of Dentistry, Department of Biomate, 歯学部, 助手 (60142444)
SATO Kazuko Tokyo Medical and Dental University, Faculty of Dentistry, Department of Biomate, 歯学部, 助手 (50046083)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | Cytotoxicity test method / SV-40 large T antigen / Human dental pulp / Mouse calvaria / Odontoblast / Osteoblast / Intracellular Ca^<2+> / Neutral red assay / 遺伝子導入 / 骨芽細胞株の樹立 / 石灰化 / 細胞内カルシウムイオン / マウス頭頂骨細胞 / ウサギ腎細胞 / SV40DNA / 培養細胞 / ヒト歯髄細胞 |
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| Research Abstract |
In order to develop a new cytotoxicity test method for assessing possible toxic effects of dental materials, the study was performed to establish new lines of cultured cells with highly susceptible to toxicants and quantitative indexes for cytotoxicity tests.
1. The human dental pulp cells and mouse calvaria cells were transfected with the SV-40 large T antigen. Four immortalized cell lines, LSC cells from human pulp and DMCs LMCs and VMCs cells from mouse calvaria were obtained. These cell lines preserved some of the properties of the dental pulp or the osteoblast.
2. The susceptibility of four cytotoxicity assays were compared. The MTT and NR assays showed higher susceptibility compared to the agar overlay assay and millipore filter method. The MTT assay showed the highest reproducibility among four tests.
3. The intracellular Ca ion concentration ([Ca^<2+>]i) of JTC-12 cells was measured employing Fura 2 AM fluorescence. Exposing cells to low concentrations of HgCl_2, which had no apparent effect on viability, produced a rapid and prolonged increase in [Ca^<2+>]i. The [Ca^<2+>]i was a useful index to detect the initial lesion of cultured cells exposed to dental materials.
4. These results suggest that the new cell lines and the [Ca^<2+>]i change may be useful in the future research for testing the toxicity of the dental materials.
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