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Role and Mechanisms of Gene Expression of Bone Sialoprotein in Periodontal Tissue Regeneration

Research Project

Project/Area Number 16592081
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Periodontal dentistry
Research InstitutionNihon University

Principal Investigator

OGATA Yorimasa  Nihon University, School of Dentistry at Matsudo, Professor, 松戸歯学部, 教授 (90204065)

Co-Investigator(Kenkyū-buntansha) SAMOTO Hiroshi  Nihon University, School of Dentistry at Matsudo, Research Assistant (Full-Time), 松戸歯学部, 助手 (50366613)
NAKAO Sumi  Nihon University, School of Dentistry at Matsudo, Research Assistant (Full-Time), 松戸歯学部, 助手 (20102577)
清水 映美  日本大学, 松戸歯学部, 講師 (80349969)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsBone sialoprotein / Extracellular matrix / Osteoblasts / Transcription factor / Transcriptional regulation / Gene promoter / Enamel matrix derivative / LPS / メカニカルストレス / インシュリン様成長因子 / リポポリサッカライド / FGF2 / cAMP / 歯周組織再生 / 石灰化 / 腫瘍壊死因子 / プロスタグランジンE2 / 静磁場
Research Abstract

BSP sialoprotein (BSP) is a glycoprotein that essentially unique to mineralize tissues. Therefore, BSP may function in the initial nucleation of hydroxyapatite crystal in de novo bone formation.
Enamel matrix derivative (EMD) has been developed as a periodontal regenerative treatment. While EMD is believed to induce regeneration of periodontal tissue, the precise mechanism is not known. Using the osteoblast like ROS 17/2.8 cells, we revealed that BSP mRNA levels increased 〜2.8-fold by EMD. In transient transfection analyses, EMD (50 μg/ml, 12 h) increased luciferase activities of pLUC4 (nt -425 to +60) and pLUC5 (nt -801 to +60), transfected into ROS 17/2.8 cells. Within the pLUC4 and 5, a homeodomain binding element (HOX) and a TGF-beta activation element (TAE) are present. Gel mobility shift assays with radiolabeled HOX and TAE ds-oligonucleotides revealed increased binding of nuclear proteins from EMD stimulated ROS 17/2.8 cells.
Lipopolysaccharide (LPS) is mediator of inflammatory responses in periodontal disease. LPS (1 μg/ml, 12 h) caused a marked reduction in BSP mRNA levels. The addition of antioxidant N-acetylcysteine (NAC ; 20mM) 30 min prior to stimulation with LPS attenuated the inhibition of BSP mRNA levels. Transient transfection analyses, revealed that LPS (1 μg/ml) suppressed expression of luciferase construct (-108 to +60), transfected into ROS17/2.8 cells. The effects of LPS were inhibited by PKA inhibitor H89 and the tyrosine kinase inhibitor, herbimycin A. Introduction of 2bp mutations in the inverted CCAAT, CRE, FRE and Pit-1 showed that the LPS effects were mediated by the CRE and FRE. Whereas the FRE and 3'-FRE DNA-protein complexes were decreased by LPS, CRE-protein complex did not change after LPS treatment. These studies show that LPS suppresses BSP gene transcription through PKA and tyrosine kinase-dependent pathways and that the LPS effects are mediated through CRE and FRE elements in the proximal BSP gene promoter.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (13 results)

All 2006 2005 2004

All Journal Article (13 results)

  • [Journal Article] Regulation of Bone Sialoprotein (BSP) Gene Transcription by Lipopolysaccharide.2006

    • Author(s)
      Naoko Kato et al.
    • Journal Title

      Journal of Cellular Biochemistry 97

      Pages: 368-379

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Effect of Chlorpromazine on Bone Sialoprotein (BSP) Gene Transcription.2006

    • Author(s)
      Yu Nakajima et al.
    • Journal Title

      Journal of Cellular Biochemistry 97

      Pages: 1198-1206

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Fibroblast Growth Factor 2 and Cyclic AMP Synergistically Regulate Bone Sialoprotein Gene Expression.2006

    • Author(s)
      Emi Shimizu et al.
    • Journal Title

      Bone (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Insulin-like Growth Factor-I Increases Bone Sialoprotein (BSP) Expression Through Fibroblast Growth Factor 2 Response Element and Homeodomain Protein-binding Site in the Proximal Promoter of the BSP Gene.2006

    • Author(s)
      Youhei Nakayama et al.
    • Journal Title

      Journal of Cellular Physiology (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Regulation of Bone Sialoprotein (BSP) Gene Transcription by Lipopolysaccharide.2006

    • Author(s)
      Naoko Kato et al.
    • Journal Title

      Journal of Cellular Biochemistry Vol.97

      Pages: 368-379

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of Chlorpromazine on Bone Sialoprotein (BSP) Gene Transcription.2006

    • Author(s)
      Yu Nakajima et al.
    • Journal Title

      Journal of Cellular Biochemistry Vol.97

      Pages: 1198-1206

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Fibroblast Growth Factor 2 and Cyclic AMP Synergistically Regulate Bone Sialoprotein Gene Exoression.2006

    • Author(s)
      Emi Shimizu et al.
    • Journal Title

      Bone (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Insulin-like Growth Factor-I Increases Bone Sialoprotein (BSP) Expression Through Fibroblast Growth Factor 2 Response Element and Homeodomain Protein-binding Site in the Proximal Promoter of the BSP Gene.2006

    • Author(s)
      Youhei Nakayama et al.
    • Journal Title

      Journal of Cellular Physiology (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amelogenin stimulates bone sialoprotein (BSP) expression through FGF2 response element and TGF-β_1 activation element in the promoter of the BSP gene.2005

    • Author(s)
      Emi Shimizu et al.
    • Journal Title

      Journal of Periodontology 76

      Pages: 1482-1489

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amelogenin stimulates bone sialoprotein (BSP) expression through FGF2 response element and TGF-β_1 activation element in the promoter of the BSP gene.2005

    • Author(s)
      Emi Shimizu et al.
    • Journal Title

      Journal of Periodontology Vol.73

      Pages: 1482-1489

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amelogenin stimulates bone sialoprotein (BSP) expression through FGF2 response element and TGF-β1 activation element in the promoter of the BSP gene.2005

    • Author(s)
      Emi Shimizu et al.
    • Journal Title

      Journal of Periodontology 76

      Pages: 1482-1489

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Nd : YAGレーザー照射が歯肉溝滲出液中の細菌性因子に与える効果2005

    • Author(s)
      増永 浩 ほか
    • Journal Title

      日本歯科保存学雑誌 (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Effecs of TNF-α on human osteosarcoma cell line Saos2-TNF-α regulation of bone sialoprotein gene expression in Saos2 osteoblast-like cells.2004

    • Author(s)
      Youhei Nakayama et al.
    • Journal Title

      Cell Biology International 28

      Pages: 653-660

    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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