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Experimental studies on peripheral nerve regeneration through the bio-nerve with 3D cultured Schwann cell

Research Project

Project/Area Number 14571401
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Orthopaedic surgery
Research InstitutionKEIO UNIVERSITY

Principal Investigator

NAKAO Yasushi  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (30188883)

Co-Investigator(Kenkyū-buntansha) NISHIWAKI Masao  KEIO UNIV., School of Medicine, Instructor, 医学部, 助手 (90296640)
TAKAYAMA Shinichiro  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (40138045)
HIROTAKA James  KEIO UNIV., School of Medicine, Assist. Professor, 医学部, 講師 (90338020)
MORISAWA Yasushi  KEIO UNIV., School of Medicine, Instructor, 医学部, 助手 (60306751)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥2,700,000 (Direct Cost: ¥2,700,000)
KeywordsSchwann cell / artificial nerve / nerve graft / 組織工学 / 再生医学 / 末梢神経 / ティッシュエンジニアリング
Research Abstract

Schwann cells were initially isolated P7 Wister rat sciatic nerve and dorsal root ganglia. The neurotrophic factor cDNA was inserted into multiple cloning site of shuttle vector. The adenoviral constructs were transfected into 293 packaging cell line. The cultured Schwann cells were infected with concentrated adenoviruses. The expression of the target neurotrophic factor gene was evaluated by Western blot analysis. Neurons from rat DRGs were cultured in a medium obtained from the transferred Schwann cell culture at 3days, 3 weeks, 8 weeks after infection. Neurite outgrowth was measured by immunostaining of axons with monoclonal antibody RT97. A 20mm nerve gap was bridged with a bioabsorbable polymer conduit containing a mixture of the transferred Schwann cells and Pluronic gel. 12 weeks later, histological evaluation was performed at the midportion of the grafted lesion. The transferred Schwann cells lysates showed the higher expression of the transgene product of 30 Kilo Dalton level by Western blot analysis. Histological evaluation demonstrated that, contrary to the poor regeneration in the control group, the genetically modified group showed not only well-regenerated axons, but also formations of vessels and epineurium-like tissue observed.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Komiyama T: "A novel technique for peripheral nerve reconstruction in the absence of an artificial conduit."J.Neuroscience Methods. 134. 133-140 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Komiyama T: "A novel technique to isolate adult Schwann cells for an artificial nerve conduit."J.Neuroscience Methods. 122. 195-200 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 栩木弘和: "シュワン細胞の3次元培養を応用した人工神経の開発"整形災害外科. 46. 247-251 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 仲尾保志: "ティッシュエンジニアリングを応用した人工神経の開発"末梢神経. 14. 7-16 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Komiyama T, Nakao Y, Toyama Y, Vacanti CA, Vacanti MP, Ronald Ignotz: "A novel technique for peripheral nerve reconstruction in the absence of an artificial conduit."Journal of Neuroscience Methods. 139. 133-140 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Komiyama T, Nakao Y, Toyama Y, Vacanti CA, Vacanti MP, Asou H.: "A novel technique to isolate adult Schwann cells for an artificial nerve conduit."Journal of Neuroscience Methods. 122. 195-200 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tochigi N, Nakao Y: "Artificial peripheral nerve using 3D culturing of Schwann cells."Seikei-saigaigeka. 46. 247-251 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nakao Y, Ikada Y: "Tissue-engineered peripheral nerve."Peripheral Nerve. 14. 7-16 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 栩木弘和, 仲尾保志: "遺伝子導入シュワン細胞を用いたハイブリッド型人工神経の開発"関節外科. 22. 98-104 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 仲尾保志: "末梢神経再生のためのTissue Engineeringの応用"脳の科学. 25. 228-233 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T Komiyama, Y Nakao: "Novel Culture Technique of Adult Schwann Cell for Hybrid Artificial Peripheral Nerve"J Neurosci Methods. 122. 195-200 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T Komiyama, Y Nakao: "A novel technique for peripheral nerve reconstruction in the absence of an artificial conduit"J Neurosci Methods. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] N Okuyama, Y Nakao: "The Effect of the Number of Fascicle on Axonal Regeneration in Cable Grafts"Microsurgery. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] 仲尾保志: "末梢神経再生のためのTissue Engineeringの応用"脳の科学. 25巻. 228-233 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Komiyama, Nakao: "Novel Culture Technique of Adult Schwann Cell for Hybrid Artificial Peripheral Nerve"J.neurosci Methods. 122. 195-200 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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