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Formation of Biotooth in Three Dimensional Culture of Dental Pulp Stem Cells Transfected with BMP

Research Project

Project/Area Number 15209065
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Conservative dentistry
Research InstitutionKYUSYU UNIVERSITY

Principal Investigator

AKAMINE Akifumi  Kyushu University, Faculty of Dental Science, Professor, 大学院・歯学研究院, 教授 (00117053)

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)
渡邊 武  九州大学, 生体防御医学研究所, 教授 (40028684)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥50,960,000 (Direct Cost: ¥39,200,000、Indirect Cost: ¥11,760,000)
Fiscal Year 2004: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
Fiscal Year 2003: ¥44,330,000 (Direct Cost: ¥34,100,000、Indirect Cost: ¥10,230,000)
KeywordsGene Therapy / Dentin / Pulp Regeneration / Biotooth / Dental Pulp Stem Cells / Side Population Cells / Odontoblasts / Bone Morphogenetic_Proteins (BMPs) / Flow Cytometry / 遺伝子導入 / バイオ歯 / 三次元培養 / BMP / scaffold
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.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (15 results)

All 2005 2004 2003 Other

All Journal Article (12 results) Publications (3 results)

  • [Journal Article] The Application of Tissue Engineering to Regeneration of Pulp and Dentin in Endodontics2005

    • Author(s)
      Nakashima M., Akamine A.
    • Journal Title

      J Endod in press

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The Application of Tissue Engineering to Regeneration of Pulp and Dentin in Endodontics2005

    • Author(s)
      Nakashima M., Akamine A.
    • Journal Title

      J Endod (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Stimulation of reparative dentin formation with ex vivo gene therapy using dental pulp stem cells electrotransfected with growth/differentiation factor 11 (Gdfl1).2004

    • Author(s)
      Nakashima M., Iohara K., Ishikawa M., Ito M., Tomokiyo A., Tanaka T., Akamine A.
    • Journal Title

      Hum Gene Ther 15

      Pages: 1045-1053

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 22004

    • Author(s)
      Iohara K., Nakashima M., Ito M., Ishikawa M., Nakasima A., Akamine A
    • Journal Title

      J Dent Res 83

      Pages: 590-595

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Stimulation of reparative dentin formation with ex vivo gene therapy using dental pulp stem cells electrotransfected with growth/differentiation factor 11(Gdf 11)2004

    • Author(s)
      Nakashima M., Iohara K., Ishikawa M., Ito M., Tomokiyo A., Tanaka T., Akamine A.
    • Journal Title

      Hum Gene Ther 15,11

      Pages: 1045-1053

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2.2004

    • Author(s)
      Iohara K., Nakashima M., Ito M., Ishikawa M., Nakasima A, Akamine A.
    • Journal Title

      J Dent Res 83,8

      Pages: 590-595

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Stimulation of reparative dentin formation with ex vivo gene therapy using dental pulp stem cells electrotransfected with growth/differentiation factor 11 (Gdf11).2004

    • Author(s)
      Nakashima M., Iohara K., Ishikawa M., Ito M., Tomokiyo A., Tanaka T., Akamine A.
    • Journal Title

      Hum Gene Ther 15・11

      Pages: 1045-1053

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 22004

    • Author(s)
      Iohara K., Nakashima M., Ito M., Ishikawa M., Nakashima A., Akamine A
    • Journal Title

      J Dent Res 83・8

      Pages: 590-595

    • Related Report
      2004 Annual Research Report
  • [Journal Article] BMPを用いた歯髄細胞導入治療法による象牙質再生2003

    • Author(s)
      庵原耕一郎, 中島美砂子, 石河真幸, 中島昭彦, 赤峰昭文
    • Journal Title

      日本歯科保存学雑誌 46

      Pages: 654-665

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Dental pulp stem cell therapy for dentin regeneration with recombinant human bone morphogenetic protein (BMP) 2.2003

    • Author(s)
      Iohara K., Nakashima M., Ishikawa M., Nakasima A, Akamine A.
    • Journal Title

      Japanese J Conservative Dentistry 46,5

      Pages: 654-665

    • NAID

      50000339659

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] BMPを用いた歯髄細胞導入治療法による象牙質再生2003

    • Author(s)
      庵原耕一郎, 中島美砂子, 石河真幸, 中島昭彦, 赤峰昭文
    • Journal Title

      日本歯科保存学雑誌 46・5

      Pages: 654-665

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The Application of Tissue Engineering to Regeneration of Pulp and Dentin in Endodontics.

    • Author(s)
      Nakashima M., Akamine A.
    • Journal Title

      J Endod (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Nakashima M., Tachibana K., IoharaK., Ito M, Ishikawa M., Akamine A.: "Induction of reparative dentin formation by ultrasound-mediated gene delivery of Growth/differentiation factor 11"Human Gene Therapy. 14. 591-597 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 庵原耕一郎, 中島美砂子, 石河真幸, 中島昭彦, 赤峰昭文: "BMP2を用いた歯髄細胞導入治療法による象牙質再生"日本歯科保存学雑誌. 46・5. 654-665 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nakashima M., Reddi A.H.: "The application of bone morphogenetic proteins to dental tissue engineering"Nature Biotech.. 21・9. 1025-1032 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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