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MUSCLE RELAXANT DERIVED FROM HUMAN GINGIVAL MICROVASCULAR ENDOTHELIAL CELLS

Research Project

Project/Area Number 07807189
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 矯正・小児・社会系歯学
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

MORITA Manabu  OKAYAMA UNIVERSITY DENTAL HOSPITAL,LECTURER, 歯学部・附属病院, 講師 (40157904)

Co-Investigator(Kenkyū-buntansha) NISHI Kazuya  OKAYAMA UNIVERSITY DENTAL HOSPITAL,ASSISTANT, 歯学部・附属病院, 助手 (10274002)
ISHIKAWA Akira  OKAYAMA UNIVERSITY DENTAL HOSPITAL,LECTURER, 歯学部・附属病院, 講師 (70222958)
WATANABE Tatsuo  OKAYAMA UNIVERSITY DENTAL SCHOOL,PROFESSOR, 歯学部, 教授 (20034176)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsGINGIVAL CAPILLARY / ENDOTHELIAL CELLS / MUSCLE RELAXANT / MECHANICAL STIMULATION / ヒト歯肉微小血管 / 弛緩因子 / 蛍光法
Research Abstract

The purpose of this investigation was to detect the smooth muscle relaxant (Nitric oxide, No) released from human gingival microvascular endothelial cells (HGECs). The effect of mechanical stimulation on NO-release from HGECs was also compared with that from human umbilical vein and artery endothelial cells. Afer collagenase treatment, human gingival samples were compressed to release endothelial cells. Centrifuged cells were resuspended in the growth medium. The cells suspension was introduced into every well of a 96-well plate. Morphologically homogeneous cells with a cobblestone appearance were harvested. The isolated cells were identified as HGECs by the presence of von Willebrand factor, the uptake of acetylated LDL,and the formation of capillary-like structure on Matrigel. Fluorometric assay was used to detect the NO.The sample was reacted with DAN reagent to form 1- (H) -naphthotiazol. After 10-min incubation at 20 ゚C,the reaction was terminated with NaOH.The end product, 2,3-diamino-naphthotriazol, was measured using a fluorescent plate reader with excitation at 360 nm and emission read at 460nm. At baseline, none of the cells tested released the NO.The NO was detected in the culture medium of the umbilical vein and artery endothelial cells after 3-day-and 5-day-mechanical stimulation by Flexercell. However, the HGECs did not release the NO during the whole experimental period. The cultivable HGECs were passage 4, which were thought not to be enough for detecting the NO.The further study will focus on more efficient method for cultivating the HGECs.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] 西 一也: "ヒト歯肉微小血管内皮細胞の分離・培養" 岡山歯誌. 14. 107-119 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 森木 大輔: "機械的刺激下におけるヒト臍帯動静脈血管内皮細胞の増殖能の違い" 岡山歯誌. 16. 107-116 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 渡邊 達夫: "ヒト歯肉毛細血管内皮細胞の機械的刺激に対する応答性" 平成7年度科学研究費補助金(一般研究A)研究成果報告書, 26 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nishi, K.: "Isolation of human gingival microvascular endothelial cells." J.Okayama Dent.Soc.107-119 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Watanabe, T.: "Response of human gingival microvascular Endothelial cells to mechanical stress" Report of Grant-in-Aid for Scientific Research (A). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Moriki, D.: "Effect of mechanical stimulation on proliferation of human umbilical endothelial cell." J.Okayama Dent.Soc.107-116 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary

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Published: 1995-04-01   Modified: 2016-04-21  

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