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Elucidation of the mechanism of collagen fiber synthesis and dissolution by fibroblast using novel multi-imaging

Research Project

Project/Area Number 20K18469
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 57010:Oral biological science-related
Research InstitutionKurume University

Principal Investigator

Hirashima Shingo  久留米大学, 医学部, 講師 (70647784)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2021: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2020: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords歯根膜 / 電子顕微鏡 / FIB-SEM / 線維芽細胞 / 超微形態解析 / 3次元解析 / 3次元超微形態解析 / Deep learning / Array tomography / コラーゲン線維 / FIB/SEM / picrosirius red染色 / CLEM / 3次元微細構造解析 / deep learning / Gap結合
Outline of Research at the Start

本研究は、in vitroにおいて線維芽細胞の細胞内-外におけるコラーゲン線維および線維束の形成/分解過程とその調節機構を、新機軸イメージング法(光-電子相関顕微鏡法・FIB/SEM tomography法・3次元免疫電顕法等)を用いて機能形態学的に明らかにすることを目的とする。また近年、ギャップ結合を介した線維芽細胞の細胞間コミュニケーションは創傷治癒やがんの浸潤・転移、臓器の線維化等に関連することが報告されており、線維芽細胞関連疾患における新たな治療標的としてギャップ結合についても検討する。

Outline of Final Research Achievements

The directionality of fiber formation of collagen fiber bundles in periodontal ligament fibroblasts was examined using CLEM combined with Picrosirius red staining and FIB-SEM. The experimental group was divided into two groups: one with gap junction inhibitor added to the culture medium and the other without, and analyzed using deep learning. The results showed that there was a significant morphological difference in the direction of fiber formation between the experimental and control groups, based on quantitative analysis of 3D reconstructions of serial electron microscopy images of fibers formed in the experimental and control groups. These results suggest that gap junctions between periodontal ligament fibroblasts may be involved in the formation of collagen fiber bundles in the periodontal ligament, especially in determining the directionality of these fibers.

Academic Significance and Societal Importance of the Research Achievements

歯根膜線維は局所で一定の方向性を持つ。しかしながら、その決定方法や線維の合成・分解の方法については明らかではない。当研究による結果は、歯根膜におけるコラーゲン線維束形成や方向性決定について、歯根膜線維芽細胞間のギャップ結合が関連している可能性を示しており、今後コラーゲン線維束の形成方法の解明や新規治療法の開発への基盤的知見となり得る。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (13 results)

All 2023 2022 2021 2020 Other

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 4 results) Presentation (6 results) (of which Int'l Joint Research: 1 results,  Invited: 5 results) Remarks (1 results)

  • [Journal Article] Mesoscopic structural analysis via deep learning processing, with a special reference to in vitro alteration in collagen fibre induced by a gap junction inhibitor2022

    • Author(s)
      Hirashima Shingo、Ohta Keisuke、Togo Akinobu、Tsuneyoshi Risa、Kusukawa Jingo、Nakamura Kei-ichiro
    • Journal Title

      Microscopy

      Volume: 72 Issue: 1 Pages: 18-26

    • DOI

      10.1093/jmicro/dfac044

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Correlative volume-imaging using combined array tomography and FIB-SEM tomography with beam deceleration for 3D architecture visualization in tissue2022

    • Author(s)
      Hirashima Shingo、Ohta Keisuke、Rikimaru-Nishi Yukiko、Togo Akinobu、Funatsu Takashi、Tsuneyoshi Risa、Shima Yuichi、Nakamura Kei-ichiro
    • Journal Title

      Microscopy

      Volume: - Issue: 3 Pages: 187-192

    • DOI

      10.1093/jmicro/dfac015

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] 3D mesoscopic architecture of a heterogeneous cellular network in the cementum?periodontal ligament?alveolar bone complex2021

    • Author(s)
      Hirashima Shingo、Ohta Keisuke、Togo Akinobu、Nakamura Kei-ichiro
    • Journal Title

      Microscopy

      Volume: 71 Issue: 1 Pages: 22-33

    • DOI

      10.1093/jmicro/dfab051

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Correlation of organelle dynamics between light microscopic live imaging and electron microscopic 3D architecture using FIB-SEM2021

    • Author(s)
      Ohta K, Hirashima S, Miyazono Y, Togo A and Nakamura K-i
    • Journal Title

      Microscopy (Oxf)

      Volume: 70 Issue: 2 Pages: 161-170

    • DOI

      10.1093/jmicro/dfaa071

    • Related Report
      2021 Research-status Report 2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Ectopic subcutaneous transplantation of fetal rat urogenital sinus and seminal vesicle promotes the organ growth and formation2020

    • Author(s)
      Uemura Kei-ichiro、Hayashi Tokumasa、Hiroshige Tasuku、Ueda Kosuke、Ohta Keisuke、Kanazawa Tomonoshin、Hirashima Shingo、Nakiri Makoto、Igawa Tsukasa、Nakamura Kei-ichiro
    • Journal Title

      Acta Histochemica

      Volume: 122 Issue: 5 Pages: 151569-151569

    • DOI

      10.1016/j.acthis.2020.151569

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Correlative imaging of collagen fibers and fibroblasts using CLEM optimized for picrosirius red staining and FIB/SEM tomography2020

    • Author(s)
      Hirashima Shingo、Ohta Keisuke、Kanazawa Tomonoshin、Togo Akinobu、Tsuneyoshi Risa、Miyazono Yoshihiro、Kusukawa Jingo、Nakamura Kei-ichiro
    • Journal Title

      Microscopy

      Volume: 69 Issue: 5 Pages: 324-329

    • DOI

      10.1093/jmicro/dfaa024

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 3D imaging analysis of cellular mesoscopic architecture.2023

    • Author(s)
      Shingo Hirashima, Keisuke Ohta, Yuichi Shima, Kei-ichiro Nakamura.
    • Organizer
      日本顕微鏡学会総会第78回学術講演 最先端電子顕微鏡法に関する国際若手シンポジウム
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 歯根膜組織構造をモデルとしたコラーゲン線維束形成および配列機構の解析.2021

    • Author(s)
      平嶋伸悟.
    • Organizer
      日本顕微鏡学会総会第77回学術講演 第22回(2021年度)日本顕微鏡学会奨励賞受賞講演.
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] In vitroにおけるコラーゲン線維配向性の定量解析.2021

    • Author(s)
      平嶋伸悟,太田啓介,金澤知之進,都合亜記暢,常吉梨沙,宮園 佳宏, 楠川仁悟,中村桂一郎.
    • Organizer
      日本顕微鏡学会総会第77回学術講演
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] Picrosirius red 染色とFIB/SEM tomographyによるCLEMを用いた線維芽細胞とコラーゲン線維の3次元微細構造解2020

    • Author(s)
      平嶋伸悟,太田啓介,金澤知之進,都合亜記暢,常吉梨沙,宮園 佳宏, 楠川仁悟,中村桂一郎
    • Organizer
      日本顕微鏡学会 第63回シンポジウム
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] 生命科学を照らす革新的イメージング技術 骨のメソスケール形態を計測する3D電子顕微鏡技術2020

    • Author(s)
      太田啓介,平嶋伸悟
    • Organizer
      第40回日本骨形態計測学会
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] CLEM (ピクロシリウスレッド染色+FIB/SEM)によるコラーゲン線維の解析2020

    • Author(s)
      平嶋伸悟,太田啓介,金澤知之進,都合亜記暢,常吉梨沙,宮園 佳宏,中村桂一郎
    • Organizer
      日本顕微鏡学会総会第76回学術講演
    • Related Report
      2020 Research-status Report
  • [Remarks] 久留米大学医学部 顕微解剖・生体形成部門

    • URL

      https://www.med.kurume-u.ac.jp/med/anat2/index.html

    • Related Report
      2022 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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