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Dye assay of matrix components in gingival crevicular fluid for destructive activity of alveolar bone

Research Project

Project/Area Number 08457514
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Conservative dentistry
Research InstitutionAsahi University

Principal Investigator

IWAYAMA Yukio  Asahi Univ.School of Dentistry, Prof., 歯学部, 教授 (70028744)

Co-Investigator(Kenkyū-buntansha) KOU Yoshitada  School of Dentistry Asahi University Instructor, 歯学部, 助手 (40298435)
SHIBUTANI Toshiaki  School of Dentistry, Asahi University Ass.Prof., 歯学部, 講師 (40206149)
柴田 麻紀  朝日大学, 歯学部, 助手 (20278203)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥4,100,000 (Direct Cost: ¥4,100,000)
Keywordsmatrix / alveolar bone / sulfated glycosaminoglycan / alcian blur / disease activity / gingival crevicular fluid / 歯肉炎
Research Abstract

A modification of the method of detecting sulfated glycosaminoglycans (S-GAG) was adapted for use in an Alcian blue assay. We used low pH,and high salt concentration to minimize interference from other polyanions such as DNA,protein, or hyaluronate. The samples of S-GAGs including chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate and heparan sulfate were blotted onto cellulose acetate strips and stained with 0.2% Alcian blue solution containing 0.05M MgCl^2,0.4M guanidine-HCl, 0.002M sulfuric acid, and 0.25% Triton X-100 at pH 1.5. There was some variability in the binding ability of S-GAG species to the dye. Chondroitin 4-sulfate showed the greatest affinity for the dye while heparan sulfate had the weakest.
The amount of dye bound by DNA,bovine serum albumin, or hyaluronate was negligible. The new method developed here can be adapted as a quantitative assay using a scanning densitometer with a linear range to 300ng of S-GAG.Moreover, this assay method can be used for direct measurement of S-GAG in the human biological fluid (saliva, serum, synovial fluid) without interferences by other macromolecular polyanion.
The total S-GAG obtained from the timed samples of gingival crevicular fluid was significantly higher (P<0.001) in the periodontitis subjects (71.41(]SY.+-。[)20.04ng) compared to the healthy (7.83(]SY.+-。[)2.21ng), and gingivitis subjects (10.53(]SY.+-。[)4.28ng).

Report

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

    (5 results)

All Other

All Publications (5 results)

  • [Publications] 山本 英治: "アルシアンブルーによる硫酸化グリコサミノグリカンの微量検出" 岐阜歯科学会雑誌. 24. 83-89 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 山本 英治: "硫酸化グリコサミノグリカンのアルシアンブルーによる微量分析-生体粗試料直接測定時における陰性荷電物質の影響-" 日本歯周病学会会誌. 40. 44-51 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] EIJI YAMAMOTO: "Micro-assay for Sulfated Glycosaminoglycans with Alcian blue" J.Gifu Dent.Soc.Vol.24, No.1. 83-89 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Eiji Yamamoto: "Microassay for Sulfated Glycosaminoglycans by Alcian blue Staining-Effects of Polyanionic Macromolecules on Staining in Untreated Biological Samples-" J.Jpn.Soc.Periodoutol.Vol40. 44-51 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 山本英治: "硫酸化グリコサミノグリカンのアルシアンブルーによる微量分析-生体粗試料における直接測定時の陰性荷重物質の影像-" 日本歯周病学会会誌. 40. 44-51 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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