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Development of Atomic Force Microscope System Combined with Confocal Laser Scanning Microscope to Simultaneously Measure Mechanical Stiffness and Observe Microstructure of Cultured Cells

Research Project

Project/Area Number 12480257
Research Category

Grant-in-Aid for Scientific Research (B)

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

Principal Investigator

SATO Masaaki  Tohoku University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (30111371)

Co-Investigator(Kenkyū-buntansha) OHASHI Toshiro  Tohoku University, Graduate School of Engineering, Research Associate, 大学院・工学研究科, 助手 (30270812)
MATSUMOTO Takeo  Tohoku University, Graduate School of Engineering, Associate Professor, 大学院・工学研究科, 助教授 (30209639)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2001: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 2000: ¥8,300,000 (Direct Cost: ¥8,300,000)
KeywordsBiomechanics / Shear stress / Endothelial Cell / Actin filament / GFP / FAT / Integrin / Finite element method / 力学的刺激
Research Abstract

Under flow condition endothelial cells are elongated and oriented to flow direction depending upon the level of shear stress and the duration of exposure. It is well known that cytoskeletal components, especially F-actin filaments, are playing important roles in the process of adaptation of cell shape to mechanical environment. In this project We have mainly focued two topics and obtained the results shown below.
1. Dynamic behavior of actin filament in living cells
We observed dynamic behavior of actin filaments in living cultured endothelial cells during exposure to shear stress. To do this, a vector of a green fluorescent protein (GFP)/actin fusion was introduced into cells using a lipofectoamine. Dynamic behavior of actin filament in an endothelial cell after introducing cytochalasin D was observed through CCD camera under an inverted fluorescent microscope. In different experiments, the endothelial cells were exposed to fluid shear stress of 2 Pa in a parallel plate flow chamber and … More the cytoskeletal structure was observed. Actin filaments located first at peripheral regions seemed to change the alignment after flow exposure and aligned to a long axis of the cell, which was not oriented to the flow direction yet, but already elongated.
2. Stress analysis in a cell exposed to shear stress
A fluid flow-structural analysis, i.e. a coupled field analysis, was performed to simulate three dimensional stress distribution in endothelial cells exposed to shear stress. The three-dimensional finite element model was generated on the basis of the cell surface geometry measured by an atomic force microscopy. The model consisted of a fluid element and a solid element representing the flow field and the endothelial cells, respectively. Analytical results on stress distribution in the cell showed that high compressive stress appeared both in the upstream side and the downstream side. These results may indicate that the stress distributions in the cells have close correlation with the F-actin distributions. Less

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] T.Ohashi: "Surface topography measurement and intracellular stress analysis of cultured endothelial cells exposed to fluid shear stress"JSME International Journal, Series C. 43. 780-786 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T.Matsumoto: "Smooth muscle cells freshly isolated from rat thoracic aortas are much stiffer than cultured bovine cells : Possible effect of phenotype"JSME International Journal, Series C. 43. 867-874 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y.Ishii: "Changes in local mechanical stiffness of cultured endothelial cells by exposure to shear stress"Proc.12th Conf of the European Society of Biomechanics. 276 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M.Sato: "Endothelial cells under flow-Regulation of cellular activity by mechanical stimuli"Proc.Jap.-Euro.Kobe Symposium on Thrombosis. 22-23 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 大橋俊朗: "有限要素法による流れ負荷培養内皮細胞の力学的解析"日本ME学会雑誌. 14. 21-24 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 佐藤正明: "細胞のバイオレオロジー -細胞の形態を決定する構造因子は?-"日本バイオレオロジー学会誌. 15. 35-37 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 佐藤正明: "せん断応力負荷と血管内皮細胞内アクチンフィラメントの再配列"第24回日本バイオレオロジー学会年会抄録集. 37 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 露木 啓: "せん断応力負荷時の単一内皮細胞内アクチンフィラメント構造変化の実時間観察"第12回バイオエンジニアリング学術講演会論文集. 85-86 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T.Ohashi: "Analysis of local mechanical properties of sheared endothelial cells measured by atomic force microscopy"Proc.ASME 2001 Summer Bioengineering Conference. BED-50. 159-160 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 大橋俊朗: "流体-構造連成解析による流れ負荷培養内皮細胞の構造最適化シミュレーション"日本数値流体力学会誌. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] N.Sakamoto: "Effect of magnetic field on nitric oxide synthesis of cultured endothelial cells"International Journal Applied Electromagnetics and Mechanics. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Ohashi: "Surface topography measurement and intracellular stress analysis of cultured endothelial cells exposed to fluid shear stress"JSME International Journal, Series C. 43 (4). 780-786 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Matsumoto: "Smooth muscle cells freshly isolated from rat thoracic aortas are much stiffer than cultured bovine cell: Possible effect of phenotype"JSME International Journal, Series C. 43 (4). 867-874 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y. Ishii: "Changes in local mechanical stiffness of cultured endothelial cells by exposure to shear stress"Proc. 12th 2th Conf of the European Society of Biomechanics. 276 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Sato: "Endothelial cells under flow - Regulation of cellular activity by mechanical stimuli -"Proc. Jap.-Euro. Kobe Symposium on Thrombosis. 22-23 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Ohashi: "Mechanical analysis of cultured endothelial cells exposed to flow by the finite element method"Jap. J. Med. Elect. Biol. Eng.. 14 (10). 21-24 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Sato: "Biorheology of endothelial cells -What structural factors determine the cell morphology?-"J. Jap. Soc. Biorheol.. 15 (2). 35-37 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Sato: "Fluid-imposed shear stress and reorganization of actin filaments in cultured endothelial cells"Proc. 24th Jap. Soc. Biorheol.. 37 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Tsuyuki: "Real time observation of change in actin structure in a single endothelial cell under fluid shear stress"Proc. 12th Bioeng Conf., JSME. 85-86 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Ohashi: "Analysis of local mechanical properties of sheared endothelial cells measured by atomic force microscopy"Proc. ASME 2001 Summer Bioengineering Conference. 159-160 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Ohashi: "Structural optimization method of sheard endothelial cells using coupled fluid-structural analysis"J. Jap. Soc. Comput. Fluid Dynamics. 9 (4) (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] N. Sakamoto: "Effect of magnetic field on nitric oxide synthesis of cultured endothelial cells."Int. J. Applied Electromag. Mech. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 斎藤正明: "せん断応力負荷と血管内皮細胞内アクチンフィラメントの再配列"第24回日本バイオレオロジー学会年会抄録集. 37 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 露木 啓: "せん断応力負荷時の単一内皮細胞内アクチンフィラメント構造変化の実時間観察"第12回バイオエンジニアリング学術講演会論文集. 85-86 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Ohashi: "Analysis of local mechanical properties of sheared endothelial cells measured by atomic force microscopy"Proc. ASME 2001 Summer Bioengineering Conference. BED-50. 159-160 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 大橋俊朗: "液体一構造連成解析による流れ負荷培養内皮細胞の構造最適化シミュレーション"日本数値流体力学会. 印刷中.

    • Related Report
      2001 Annual Research Report
  • [Publications] N.Sakamoto: "Effect of magnetic field on nitric oxide synthesis of cultured endothelial cells"International Journal Applied Electromagnetics and Mechanics. 印刷中.

    • Related Report
      2001 Annual Research Report
  • [Publications] 佐藤正明: "細胞のバイオテオロジー -細胞の形態を決定する構造因子は?-"日本バイオレオロジー学会誌. 15・2. 35-37 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Ohashi: "Surface topography measurement and intracellular stress analysis of cultured endothelial cells exposed to fluid shear stress"JSME International Journal, Series C. 43・4. 780-786 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Sato: "Mechanical stress and actin filament dynamics in cultured endothelial cell"Proc.4th Int.Symp.on Tissue Engineering for Therapeutic Use 4. 121-130 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 大橋俊朗: "有限要素法による流れ負荷培養内皮細胞の力学的解析"日本ME学会雑誌BME. 14・10. 21-24 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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