Analysis of function of DMP-1 during reparative dentinogenesis and its application for dentin regenerative therapy
| Project/Area Number |
17591989
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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 |
Conservative dentistry
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| Research Institution | Osaka University |
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Principal Investigator |
NOGUCHI Nobuo (2006) Osaka University, Dental Hospital, Resident, 歯学部附属病院, 医員 (80423143)
河原 敬 (2005) 大阪大学, 歯学部附属病院, 医員 (90397646)
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| Co-Investigator(Kenkyū-buntansha) |
TOYOSAWA Satoru Osaka University, Graduate School of Dentistry, Professor, 大学院歯学研究科, 教授 (30243249)
UEDA Mio Osaka University, Dental Hospital, Resident, 歯学部附属病院, 医員 (90423136)
野口 展生 大阪大学, 歯学部附属病院, 医員 (80423143)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | DMP-1 / Osteopontin / Osteocalcin / Odontoblast / Reparative Dentin / Regeneration of Dentin / osteopontin |
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| Research Abstract |
To clarify the functional role of DMP-1 during reparative dentinogenesis, the following three experiments were performed.
1. The expression of Osteopontin (OPN) and Osteocalcin (OCN) was evaluated after cavity preparation on rat molars using immunohistochemistry. OPN was expressed 7 days after cavity preparation along the dentinal tubules beneath the cavity, and continuously observed until day 28. OCN was detected not only beneath the cavity prepared but also in whole dentin from day 0 to 28.
2. We examined the chronological gene expression of DMP-1 and OPN in the pulpal cells was by real-time RT-PCR methods. The tooth was extracted and total RNA was extracted from the pulpal cells after cavity preparation, and then mRNA expression of DMP-1 and OPN was quantitatively measured using comparative Ct methods. The mRNA expression of DMP-1 and OPN was found to be increased in advance the immunohistochemical appearance of proteins.
Increased mRNA expression of DMP-1 and OPN was observed in reaction to the stimulation to the pulp by cavity preparation, and DMP-1 and OPN were found to be secreted following expression of mRNA of these molecules in the process of reparative dentin formation. These findings suggest that DMP-1 and OPN may play some important functional role in the healing process of pulp tissues.
3. We attempted to produce recombinant DMP-1. After GST-fusion protein was created in Escherichia coli expression construct vector, recombinant DMP-1 was purified by affinity chromatography on GST beads. As it proved that purified DMP-1 was degraded and fragmented using SDS-PAGE, purification condition was modified and we successfully obtained the recombinant DMP-1.
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Report
(3 results)
Research Products
(2 results)
