2004 Fiscal Year Final Research Report Summary
Formation of Biotooth in Three Dimensional Culture of Dental Pulp Stem Cells Transfected with BMP
| Project/Area Number |
15209065
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | KYUSYU UNIVERSITY |
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Principal Investigator |
AKAMINE Akifumi Kyushu University, Faculty of Dental Science, Professor, 大学院・歯学研究院, 教授 (00117053)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASHIMA Misako Kyushu University, Faculty of Dental Science, Research Associate, 大学院・歯学研究院, 助手 (20207773)
SHIRAHATA Sanetaka Kyushu University, Faculty of Agriculture, Professor, 大学院・農学研究院, 教授 (90154377)
HIRATA Masako Kyushu University, Hospital, Research Associate, 大学病院, 助手 (10153769)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Gene Therapy / Dentin / Pulp Regeneration / Biotooth / Dental Pulp Stem Cells / Side Population Cells / Odontoblasts / Bone Morphogenetic_Proteins (BMPs) / Flow Cytometry |
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| Research Abstract |
In this investigation, we have tried to develop a biotooth with dentin-pulp complex in vitro using pulp stem cells transfected with Bmp. Side population (SP) cells were isolated from adult pulp tissue based on the exclusion of the DNA dye Hoechst 33342. The more enriched stem cell fractions were obtained after staining with CD117, CD90, CD31 and CD135. They exhibited stem cell activity, such as self-renewal capability and multipotency to differentiate into neurons, chondrocytes and adipocytes as well as odontoblasts. The expression of odontoblast differentiation markers was increased in the pulp stem cells transfected with Bmp compared with control pCAGGS. The Bmp transfected cells were cultured on the scaffold with concaved shape and further layered by pulp stem cells. The odontoblast-like cells attaching to the scaffold extended long cellular processes into the scaffold seven days afterr cultivation. Dspp mRNA was expressed in the odontoblastic layers, suggesting the formation of dentin-pulp complex. The in vivo evaluation in the dog was undertaken using this biotooth. Transplantation of the biotooth on amputated pulp induced regenerative dentin formation. In conclusion, the pulp stem cells transfected with Bmp combined with the proper scaffold can be used for regenerative therapy of dentin/pulp complex for endodontic treatment.
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