| Project/Area Number |
11470406
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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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 | KYUSHU UNIVERSITY |
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Principal Investigator |
NAKASHIMA Misako Faculty of Dental Science KYUSHU UNIVERSITY Assistant Professor, 大学院・歯学研究院, 助手 (20207773)
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| Co-Investigator(Kenkyū-buntansha) |
GOTO Yasuharu Faculty of Dental Science, KYUSHU UNIVERSITY Assistant Professor, 大学院・歯学研究院, 助手 (00170473)
HIRATA Masako University Dental Hospital, KYUSHU UNIVERSITY Assistant Professor, 歯学部・附属病院, 助手 (10153769)
AKAMINE Akifumi Faculty of Dental Science, KYUSHU UNIVERSITY Professor, 大学院・歯学研究院, 教授 (00117053)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥15,200,000 (Direct Cost: ¥15,200,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥13,800,000 (Direct Cost: ¥13,800,000)
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| Keywords | Reparative Dentinogenesis / Gene Therapy / Bone Morphogenetic Proteins / Growth / DifferentiationFactor 11 / Dental Pulp Capping / Electroporation / Green Fluorescent Protein / Ultrasound / differentiation factor 11 |
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| Research Abstract |
Gene therapy is now evolving rapidly and gaining significant momentum as a therapeutic strategy to treat patients suffered from inherited disease, infectious disease, and malignant tumors in the near future. Among the nonviral techniques for gene transfer, electroporation is simple, inexpensive and safe. We investigated the applicability of in vivo electroporation of gene transfer into amputated dental pulp, using plasmid DNA expressing Growth/Differentiation factor ( Gdf) 11 as the vector. The bead soaked with recombinant GDF11 could induce the mRNA expression of Dentin sialoprotein (DSP), the differentiation marker of odontoblasts, in the dental papillae cells surrounding them, 7 days after organ culture. The conditions required for optimal in vitro (or ex vivo) electroporation were determined by green fluorescent protein (GFP) expression after the gene transfer of GDF11-GFP cDNA plasmid into dental papillae mesenchyme. The optimal Gdf11 gene transfer into the dental pulp mesenchymal cells much increased the mRNA expression of DSP locally as the recombinant GDF11 application. It demonstrated that Gdf11 cDNA plasmid transferred by electroporation might have potential for gene therapy of dental pulp cell differentiation into odontoblasts and reparative dentin formation in vivo. Therefore, for in vivo electroporation we used the same condition and the same electrode as the in vitro electroporation. There was a difficulty, however, in the approach of the electrode to the surface of the exposed pulp in the narrow cavity without any damage on the pulp tissue. The pressure and heat by the electrode resulted in the necrosis of the pulp tissue locally attached to the electrode. Further appraisal of nonviral mean such as ultrasound should be waited for more powerful and convenient gene transfer into the exposed dental pulp.
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