| Project/Area Number |
13671490
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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 |
Orthopaedic surgery
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| Research Institution | Chiba University |
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Principal Investigator |
YAMAZAKI Masashi Chiba University, University Hospital, Assistant, 医学部附属病院, 助手 (50281712)
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| Co-Investigator(Kenkyū-buntansha) |
SEKI Naohiko Chiba University, Graduate School of Medicine, Associate Professor, 大学院・医学研究院, 助教授 (50345013)
OKAWA Akihiko Chiba University, Graduate School of Medicine, Assistant, 大学院・医学研究院, 助手 (30312945)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | cDNA microarray / gene expression / mouse / bone formation / fracture repair / periostin / knock-out mouse / TNF-alpha / 骨折治癒 / 遺伝子 / cDNAマイクロアレイ / periostin(ペリオスチン) / in situ hybridization / Northern blotting / RT-PCR / 仮骨 |
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| Research Abstract |
To comprehensively evaluate gene expression in the early stage of fracture healing, we used a cDNA microarray with 2304 cDNA clones derived from an oligo-capped mouse embryo library. Closed middiaphyseal fractures were created in mouse tibiae and expression profiles were analyzed 3 days after fracture. The expression levels of six genes were up-regulated in comparison to those in unfractured bones, and 3 of 6 were identified as novel candidate genes involved in fracture repair. These are periostin, calumenin, and FHL-1, for which cloning has been already completed, but their function during bone formation remains to be elucidated. We also found that the expression of 11 genes was down-regulated. The up-regulation of the 6 genes was reconfirmed by semi-quantitative RT-PCR analysis. Furthermore, among the 6 genes, we analyzed the spatial and temporal expression of periostin by means of in situ hybridization and Northern blot analysis since it displayed the highest up-regulation ratio. A strong signal for periostin was detected in undifferentiated mesenchymal cells and immature osteoblastic cells in the periosteum between days 3 and 7, but the signal rapidly decreased by day 14. The temporal expression pattern exhibited a peak in expression on day 3, followed by a rapid decrease in expression on day 14. These findings suggest that periostin is a specific marker for preosteoblasts and plays an important role in callus formation during the early stage of fracture healing.
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