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Cloning, Expression and Identification of Specific Genes related Osseointegration to Titanium.

Research Project

Project/Area Number 15390592
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 補綴理工系歯学
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

KANYAMA Manabu  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor, 医学部・歯学部附属病院, 講師 (90294420)

Co-Investigator(Kenkyū-buntansha) KUBOKI Takuo  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (00225195)
TAKIGAWA Masaharu  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (20112063)
ARAKAWA Hikaru  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (30304314)
SUZUKI Kouji  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 医学部・歯学部附属病院, 助手 (30304322)
FUJISAWA Takuo  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (20325096)
中西 徹  岡山大学, 大学院・医歯学総合研究科, 助教授 (30243463)
Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥10,800,000 (Direct Cost: ¥10,800,000)
Keywordsbone marrow cells / Osteoblasts / Titanium / Osseointegration / xab-2 / sod-1 / galection-1 / デンタルインプラント / チタン特異的遺伝子 / 骨芽細胞 / 骨組織再生 / 幹葉系幹細胞
Research Abstract

The aim of this study is to clarify the cell behavior induced by titanium and to isolate specific genes that promote the titanium implant-bone integration. Especially, to identify the master key genes related osseointegration to titanium, the osteoblastic cell line, MC3T3-E1 cells were cultured on titanium coated glasses and the gene expression were analyzed.
1)Titanium and chrome coated glasses
Ti and Cr were coated to polished glass surfaces (33 mm in diameter by 1.5 mm in thickness) in a high vacuum condition. Ti and Cr coated glass disks were placed in a 6 well plate and fixed by silicone rings. Non-coated glass disks were served as controls.
2)Cell attachment, proliferation and differentiation assessments.
Initial cell attachment and proliferation of MC3T3-E1 cells were estimated by MTS-assay (CellTiter 96【○!R】 AQueous One Solution, Promega, USA), and cell differentiation was evaluated by alkaline phosphatase activity assay. The mean relative amount of the attached cells on the Ti-coa … More ted glass was significantly higher than those on non-coated and Cr-coated glass at 3 hours after seeding. On the same way the mean cell number on Ti-coated glass at 3 days after seeding was significantly higher than those of other conditions. The mean alkaline phosphatase(ALP) activity of the seeded osteoblasts also increased with time in these three conditions, while the ALP activity level on the Ti-coated glass at 14 days after seeding was significantly higher than those of other conditions. These results suggest that, Ti-coated glass plate accelerated the cell attachment, proliferation and differentiation of the MC3T3-E1 cells, compared to the non- and Cr-coated glass plates
3)Effect of gene expression of osteoblast on titanium
Osteoblasts were cultured on Ti coated, Cr coated and non-coated glass plates. And then differentially expressed genes were identified by cDNA subtractive hybridization. Twenty independent clones were isolated and by nucleotide sequencing of these clones, seven clones were identified including EST genes ; xab-2,sod-1,galectin-1,actin related protein 2/3 mRNA, RIKEN cDNA 2210013021 gene, EST 601086505F1, and EST 01439. And galectin-1,xab-2,sod-1 gene expression on Ti-coated glass were higher than non-coated, Cr-coated glass.
These results suggest that Ti-coating is more advantageous for osteoblastic cell attachment, proliferation and differentiation than Cr or non-coating and galectin-1,xab-2 and sod-1 up-regulation could be related to the Ti-bone integration. Less

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (16 results)

All 2006 2005 Other

All Journal Article (16 results)

  • [Journal Article] Effect of Titanium on the Attachment, Proliferation, Differentiation, and Gene Expression of Osteoblastic Cell Line, MC3T3-E1 cells2006

    • Author(s)
      K.AkiyamaV, F.Myokai, T.fujisawa, Y.Yosida, M.Ono, M.Kanyama, S.Takashiba, K.Suzuki, T.Kuboki
    • Journal Title

      Academy of Osseointegration 21^s^t Annual Meeting Program guide

      Pages: 80-80

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of Titanium on the Attachment, Proliferation, Differentiation, and Gene Expression of Osteoblastic Cell Line, MC3T3-E1 cells.2006

    • Author(s)
      K.Akiyama, F.Myokai, T.Fujisawa, Y.Yosida, M.Ono, M.Kanyama, S.Takashiba, K.Suzuki, T.Kuboki
    • Journal Title

      Academy of Osseointegration 21^<st> Annual Meeting Program Guide 80

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 骨芽細胞様細胞株(MC3T3E1)細胞の細胞接着・増殖・分化および遺伝子発現に対するチタンの影響2005

    • Author(s)
      秋山 謙太郎
    • Journal Title

      岡山歯学会雑誌 24(1)

      Pages: 1-16

    • 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] 再生医療と歯科補綴学の接点 補綴治療における再生医療のニーズと現時点での研究動向 -確実かつ質の高いインプラント治療を目指して-2005

    • Author(s)
      窪木拓男
    • Journal Title

      日本綴学歯科学会雑誌 49・4

      Pages: 576-586

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 骨芽細胞様細胞株(MC3T3E1)細胞の細胞接着・増殖・分化および遺伝子発現に対するチタンの影響2005

    • Author(s)
      秋山謙太郎, 藤沢拓生, 明貝文夫, 完山学, 古田靖弘, 高柴正悟, 鈴木一臣, 窪木拓男
    • Journal Title

      第35回(社)日本口腔インプラント学会総会・学術大会プログラム・抄録集

      Pages: 85-85

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Initial cell attachment, proliferation and differentiation of osteoblast like(MC3T3-Ei) cells to titanium chrome2005

    • Author(s)
      Akiyama K, Fujisawa T, Ono M, Yoshida Y, kanyama M, Suzuki K, KUboki T
    • Journal Title

      The 1^s^t Congress of Asian Academy of Osseointegration Program and Abstract

      Pages: 87-87

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 補綴治療における再生医療のニーズと現時点での研究動向 -確実かつ質の高いインプラント治療を目指して-2005

    • Author(s)
      窪木拓男
    • Journal Title

      第26回岡山歯学会総会抄録集

      Pages: 3-3

    • 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 titanium on the attachment, proliferation, differentiation, and gene expression of osteoblast like cells (MC3T3-E1).2005

    • Author(s)
      K.Akiyama
    • Journal Title

      The Journal of Okayama Dental Society 24(1)

      Pages: 1-16

    • 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 titanium on the attachment, proliferation, differentiation, and gene expression of osteoblast like cells (MC3T3-E1).2005

    • Author(s)
      K.Akiyama, T.Fujisawa, F.Myokai, M.Kanyama, Y.Yoshida, S.Takashiba, K.Suzuki, T.Kuboki
    • Journal Title

      The 1^<st> Congress of Asian Academy of Osseo-integration Program and Abstract 87

    • 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 Titanium on the Attachment, Proliferation, Differentiation, and Gene Expression of Osteoblastic Cell Line, MC3T3-E1 cells.2005

    • Author(s)
      K.Akiyama, F.Myokai, T.Fujisawa, Y.Yosida, M.Ono, M.Kanyama, S.Takashiba, K.Suzuki, T.Kuboki
    • Journal Title

      The Japanese Society of Implantrogy 35^<th> Meeting Special Issue 85

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Needs and Current Research Directions of Biological Regenerative Medicine in Prosthodontic Practice -To Attain Reliable and Sophisticated Dental Implant Therapy-2005

    • Author(s)
      T.Kuboki
    • Journal Title

      The Journal of Japan Prosthodontic Society 49(4)

      Pages: 576-586

    • NAID

      10029761695

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Needs and Current Research Directions of Biological Regenerative Medicine in Prosthodontic Practice -To Attain Reliable and Sophisticated Dental Implant Therapy-2005

    • Author(s)
      T.Kuboki
    • Journal Title

      The Journal of Okayama Dental Society Special Issue

    • NAID

      10029761695

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 再生医療と歯科補綴学の接点 補綴治療における再生医療のニーズと現時点での研究動向 -確実かつ質の高いインプラント治療を目指して-2005

    • Author(s)
      窪木拓男
    • Journal Title

      日本補綴歯科学会雑誌 49・4

      Pages: 576-586

    • Related Report
      2005 Annual Research Report
  • [Journal Article] 骨芽細胞様細胞株(MC3T3E1)細胞の細胞接着・増殖・分化および遺伝子発現に対するチタンの影響2005

    • Author(s)
      秋山謙太郎, 藤澤拓生, 明貝文夫, 完山学, 吉田靖弘, 高柴正悟, 鈴木一臣, 窪木拓男
    • Journal Title

      第35回(社)日本口腔インプラント学会総会・学術大会プログラム・抄録集

      Pages: 85-85

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Initial cell attachment, proliferation and differentiation of osteoblast like (MC3T3-Ei) cells to titanium and chrome2005

    • Author(s)
      Akiyama K, Fujisawa T, Ono M, Yoshida Y, Kanyama M, Suzuki K, Kuboki T
    • Journal Title

      The 1^<st> Congress of Asian Academy of Osseointegration Program and Abstract

      Pages: 87-87

    • Related Report
      2005 Annual Research Report
  • [Journal Article] 骨芽細胞様細胞株(MC3T3-E1細胞)の細胞接着,増殖,分化および遺伝子発現に対するチタンの影響

    • Author(s)
      秋山 謙太郎
    • Journal Title

      岡山歯学会雑誌 (印刷中)

    • Related Report
      2004 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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