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Functional analyses of novel collagen formed network-matrix and its application for cell culture

Research Project

Project/Area Number 16500307
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biomedical engineering/Biological material science
Research InstitutionKinki University

Principal Investigator

MORIMOTO Koichi  Kinki University, Faculty of Biology-Oriented Science and Technology, Assistant Professor, 生物理工学部, 講師 (10319741)

Co-Investigator(Kenkyū-buntansha) SAITO Takuya  Kinki University, Faculty of Biology-Oriented Science and Technology, Associate Professor, 生物理工学部, 助教授 (30101398)
YOSHIKAWA Takafumi  Nara Medical University, Department of Orthopaedic Surgery, Assistant Professor, 整形外科, 講師 (90275347)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsCollagen / Cell culture / Cell adhesion / Neutrophil / Fibroblast 3T3-L1 / Immunoelectron microscopy / Fluorescence microscopy / Scanning electron microscopy / 微細網目形成コラーゲン / 細胞浸潤 / 3本螺旋構造 / 止血効果
Research Abstract

Type I collagen is the most abundant component in tissues such as skin and assembles spontaneously to fibrils in vitro in physiological conditions. It has been known that collagen has biological functions as well as mechanical functions.
Methods : Atelocollagen and AP-collagen were purified from chicken skin by pepsin and actinidain limited digestion, respectively. The purified collagen species were identified by 5% SDS-polyacrylamide gel electrophoresis. Morphological difference between two collagen matrices was observed by an S-900 (Hitachi) ultra-high resolution scanning electron micrograph (SEM). By using anti-collagen monoclonal antibody (COL-1), we compared immunoreactivities against two collagen matrices. We examined each polypeptide fragment digested by collagenase to obtain further evidence for the ultra-structural difference. In addition, to evaluate whether two collagen matrices may lead to recruitment of neutrophils, we observed biological behaviors of mouse neutrophil on the collagen matrices by SEM.
Results : The SDS-PAGE pattern showed that the atelocollagen consists of at least four components : α1, α2, β, and γ chain. In contrast, the AP-collagen gave two bands corresponding to α1 and α2 chain. The immunoreactivities of COL-1 to two collagen matrices were different from at pH 4.0 and at pH 6.5 conditions. Digested fragments of two collagen matrices showed different pattern in SDS-PAGE. It seemed that the adhesive and the migration activity of neutrophil were clearly different between two collagen matrices.
Conclusion : The AP-collagen formed the unique matrix such as three dimensional meshwork layers. Additional lines of evidence of the recognition sites of COL-1 antibody and collagenase against two collagen matrices strongly indicated difference in the ultra-structure. We demonstrated that the AP-collagen matrix clearly enhances the neutrophil adhesion and migration activity.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (12 results)

All 2005 2004

All Journal Article (12 results)

  • [Journal Article] アクチニダイン酵素処理にて生じたI型コラーゲンの生化学的特性の変化2005

    • Author(s)
      國井沙織
    • Journal Title

      近畿大学 先端技術総合研究所紀要 10

      Pages: 19-28

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Comparison of biochemical properties of novel actinidain-hydrolyzed collagen and pepsin-hydrolyzed collagen.2005

    • Author(s)
      Saori Kunii
    • Journal Title

      Mem.Institute of Advanced Technology, Kinki University 10

      Pages: 19-28

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] アクチニダイン処理I型コラーゲン会合体の高次構造差異から生じる物性変化2005

    • Author(s)
      國井沙織, 斎藤卓也, 柴野三智子, 森本康一
    • Journal Title

      第5回日本蛋白質科学会年会プログラム・要旨集

      Pages: 110-110

    • Related Report
      2005 Annual Research Report
  • [Journal Article] 微細網目形成I型コラーゲン・マトリックスと好中球との相互作用の走査型電子顕微鏡による時間依存観察2005

    • Author(s)
      國井沙織, 柴野三智子, 斎藤卓也, 森本康一
    • Journal Title

      日本顕微鏡学会第61回学術講演会講演概要集

      Pages: 65-65

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Ultra-structural findings of type I collagen matrix generated by actinidain limited-hydrolysis2005

    • Author(s)
      S.Kunii, M.Shibano, T.Saito, K.Morimoto
    • Journal Title

      日本生化学会誌 77・8

      Pages: 969-969

    • Related Report
      2005 Annual Research Report
  • [Journal Article] コラーゲン会合体の高次構造変化に起因する好中球細胞の新奇な運動性2005

    • Author(s)
      國井沙織, 柴野三智子, 斎藤卓也, 森本康一
    • Journal Title

      日本生物物理学会誌 45・1

    • Related Report
      2005 Annual Research Report
  • [Journal Article] ペプチダーゼ処理により生じるコラーゲン会合体の微細構造と高次構造の変化2005

    • Author(s)
      森本康一, 國井沙織, 柴野三智子, 斎藤卓也
    • Journal Title

      日本生物物理学会誌 45・1

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Difference of structural features between actinidain-hydrolyzed collagen matrices and pepsin-hydrolyzed collagen matrices2005

    • Author(s)
      S.Kunii, M.Shibano, T.Saito, K.Morimoto
    • Journal Title

      2005環太平洋国際化学会(PACIFICHEM2005)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] アクチニダイン酵素処理にて生じたI型コラーゲンの生化学的特性の変化2005

    • Author(s)
      國井 沙織 他
    • Journal Title

      近畿大学 先端技術総合研究所紀要 (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Preparation and structural analysis of actinidain-processed atelocollagen of Yellowfin tuna(Thunnus albacares)2004

    • Author(s)
      Koichi Morimoto
    • Journal Title

      Biosci. Biotechnol. Biochem. 68(4)

      Pages: 861-867

    • NAID

      10013143843

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Preparation and structural analysis of actinidain-processed atelocollagen of Yellowfin tuna (Thunnus albacares)2004

    • Author(s)
      Koichi Morimoto
    • Journal Title

      Biosci.Biotechnol.Biochem. 68(4)

      Pages: 861-867

    • NAID

      10013143843

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Preparation and structural analysis of actinidain-processed atelocollagen of Yellowfin tuna (Thunnus albacares)2004

    • Author(s)
      Koichi Morimoto, et al.
    • Journal Title

      Biosci.Biotcchnol.Biochem. 68(4)

      Pages: 861-867

    • NAID

      10013143843

    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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