Development of an osteoporosis model pro-culture by immunity restraint agent and molecular biology analysis of bone absorption mechanism
| Project/Area Number |
15592108
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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 |
Surgical dentistry
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
FUKUNAGA Jyoji Okayama University, Graduate School of Medicine and Dentistry, Assistant, 大学院・医歯学総合研究科, 助手 (10284069)
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| Co-Investigator(Kenkyū-buntansha) |
TSUJIGIWA Hidetugu Okayama University, Graduate School of Medicine and Dentistry, Assistant, 大学院・医歯学総合研究科, 助手 (70335628)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | osteoporosis / Immunosuppresant agents / osteoclast / T, B細胞 / 骨代謝マーカー / FACS / T細胞 / 3次元マイクロCT |
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| Research Abstract |
Blood and urine analysis : Although the level of serum calcium remained mostly within the normal range for both groups, there was a significant increase in the level of urinary Ca for the FK506 treated group from Week 1 of administration. The level of blood osteocalcin for the FK506 treated group at Week 1 of administration was significantly higher than that for the control group, but there was no significant difference after that. Throughout the administration period, the sum of urinary PYD and Dpd for the FK506 treated group was significantly higher. Of the various bone resorption factors, the level of blood PTH for the FK506 treated group was significantly higher than that for the control group at Week 3 of administration and later.
Cell culture : After acclimatization for a week, mice were randomly divided into two groups and were intraperitoneally injected every day either FK506 at a dose of 1mg/kg/day (FK506 treated group) or physiological saline at 10ml/kg/day (Control group). Pr
… More
ograf, a solution containing 5mg FK506/ml, was diluted in physiological saline to obtain a final concentration. bone marrow cells were collected by flushing femoral shafts using sterile needles. Cells plated in 24-well plates were cultured for 6 days in α-minimal essential medium supplemented with 10% fetal bovine serum, 100U/ml penicillin, and 100 μg/ml streptomycin, 30 ng/ml macrophage-colony stimulating factor, and 100 ng/ml soluble Receptor Activator of NFκB Ligand. TRAP assay : Staining for tartrate-resistant acid phosphatase(TRAP) activity, a leukocyte acid phosphatase assay kit was used. At the end of the culture period, TRAP positive cells exhibiting ≧3 nuclei were counted as mature osteoclasts and TRAP positive cells with <3 nuclei were considered osteoclast precursors. Resorption pit assay : Osteoclasts were seeded onto dentine slices placed in each well of 24-well plates, and after the cells were scraped off from the dentine slices, excavated pits were stained with acid hematoxylin Photographs were taken under a light microscope at ×40 magnification, and total areas of resorption pits were analyzed by the computer soft of NIH Image. Less
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Report
(3 results)
Research Products
(14 results)
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[Journal Article] Expression of Osteoclast Differentiation Factor (RANKL) and Osteoclastogenesis Inhibitory Factor (OPG) in Rat Osteoporosis Induced by Immunosuppressant FK506.2004
Author(s)
Jyoji Fukunaga, Yuichirou Yamaai, Eiki Yamachika, Yuzou Ishiwari, Hidetugu Tsujigiwa, Koichi Sawaki, Y.J.Lee, Takaaki Ueno, Shiho Kirino, Nobuyoshi Mizukawa, Shin Takagi, Noriyuki Nagai, Toshio Sugahara
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Journal Title
Description
「研究成果報告書概要(欧文)」より
Related Report
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