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Dinamic and functional analysis of osteoclast activity using video-enhanced contrast-differential interference contrast (VEC-DIC) microscopy

Research Project

Project/Area Number 17591561
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Orthopaedic surgery
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

HOSHINO Hironobu  Hamamatsu University School of Medicine, hospital affiliated to a medical school, assistant professor (70293636)

Co-Investigator(Kenkyū-buntansha) NAGANO Akira  Hamamatsu University School of Medicine, medical school, professor (50272547)
TERAKAWA Susumu  Hamamatsu University School of Medicine, photon medical research center, Professor (50014246)
Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordsosteoclast / video microscopy / pbase contrast microscope / calcitonin / bisphosphonate
Research Abstract

Quantitative evaluation of the ability of bone resorption activity in live osteoclast-like cells (OCLs) has not reported yet. To analyze the ability of bone resorption in the same alive cells for long periods, we observed OCLs maintaining normal culture condition, and recorded by video: microscopy. The coverslips were placed in the incubator equipped inverted microscope (Biostation IM, Nikon, Japan), and OCLs were observed with phase contrast microscope. We observed the morphological change of OCLs and measured calcium phosphate free area made by OCLs affected by elcatonin using incubator facilitated video-enhanced microscopy. OCLs, which were obtained from co-culture of ddy-mouse osteoblastic cells and bone marrow cells, were cultured on calcium phosphate (CP)-coated quartz coverslips to observe sequential morphological change and to measure the increase of CP-free area. The CP-free area increase constantly in the control group, while that didn't increase after the addition of agent in the elcatonin group. In this study we could evaluate the resorbed areas increased on the same CP-coated coverslips, even with the migrated osteoclast. Furthermore, we observed the morphological change of OCLs after elcatonin addition. The nuclei were observed above the CP-free area and were gathered each other before chemical addition. After addition of elcatonin, however, the vacuoles spread from CP-free area, and the nuclei were dispersed. These results suggest that elcatonin could not only weaken the bone resorption ability but also change the polarization of nuclei. This study showed that analysis of the resorbed areas under the osteoclast-like cell body using this method enables the quantitative evaluation of the bone resorption activity to the several therapeutic agents like bisphosphonates in vitro.

Report

(4 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (6 results)

All 2007 Other

All Journal Article (3 results) (of which Peer Reviewed: 2 results) Presentation (3 results)

  • [Journal Article] Mechanical fragmentation and transportation of calcium phosphate substrate by filopodia and lamellipodia in a mature osteoclast2007

    • Author(s)
      Nagafusa T, Hoshino H, Sakurai T, Terakawa S, Nagano A
    • Journal Title

      Cell Biology International 31

      Pages: 1150-1159

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Mechanical fragmentation and transportation of calcium phosphate substrate by filopodia and lamellipodia in a mature osteoclast2007

    • Author(s)
      Nagafusa, T., Hoshino, H., Sakurai, T., Terakawa, S., Nagano, A
    • Journal Title

      Cell Biology International 31

      Pages: 1150-1159

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Mechanical fragmentation and transportation of calcium phosphate subs trate by filopodia and lamellipodia in a mature osteoclast2007

    • Author(s)
      Nagafusa T, Hoshino H, Sakurai T, Terakawa S, Nagano A
    • Journal Title

      Cell Biology International 31

      Pages: 1150-1159

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] ビデオマイクロスコピーによる破骨細胞の動態機能解析2007

    • Author(s)
      森本祥隆
    • Organizer
      第22回日本整形外科学会基礎学術集会
    • Place of Presentation
      浜松市
    • Year and Date
      2007-10-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ビデオマイクロスコピーによる破骨細胞の動憲機能解析2007

    • Author(s)
      森本 祥隆
    • Organizer
      第22回日本整形外科学会基礎学術集会
    • Place of Presentation
      浜松市
    • Year and Date
      2007-10-25
    • Related Report
      2007 Annual Research Report
  • [Presentation] Dinamic and functional analysis of osteoclast activity using video microscopy

    • Author(s)
      Yoshitaka, Morimoto
    • Organizer
      The 22nd basic research conference of Japanese Orthopaedic Association
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2005-04-01   Modified: 2016-04-21  

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