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Creation and clinical application of the three-dimensional cultured cell transplantation for hard tissue regeneration

Research Project

Project/Area Number 17591996
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

IKEDA Takeshi  Nagasaki University, University Hospital of Medicine and Dentistry, Instructor, 医学部・歯学部附属病院, 助手 (90244079)

Co-Investigator(Kenkyū-buntansha) YNAGIGUCHI Kajirou  Nagasaki University, University Hospital of Medicine and Dentistry, Assistant professor, 医学部・歯学部附属病院, 講師 (50264255)
OHARA Naoko  Nagasaki University, Graduate School of Biomedical-Sciences, Instructor, 大学院医歯薬学総合研究科, 助手 (80301365)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordschitosan / hard tissue regeneration / bone marrow-derived progenitor cells / cDNA microarray / ALP / osteocalcin / BMP- / real-time PCR / 培養骨芽細胞
Research Abstract

The present study was designed to investigate histochemically the biodegradation processes of chitin and chitosan implanted in rat alveolar bone. Lysozyme was immunohistochemically detected using postembedding immunogold labeling. The degradation process was ultrastructurally observed using the lectin-colloidal gold technique with electron microscopy. Three groups of chitin were specially prepared according to their degree of deacetylation : 100% deacetylated chitin (DDAC 100) ; 50% (DDAC 50) ; and 0% (DDAC 0). The present immunohistochemical study indicated that lysozyme expression was not detected in the DDAC 100 group. Furthermore, electron microscopy clearly demonstrated that the contour of implanted chitosan changed over time, and that chitosan-like fragments were present in the phagosomes in the DDAC 50 and 100 groups. These findings strongly suggest that phagocytes, such as multinuclear cells, are easily supplied in bone tissue and that the phagocytosis is more effective than en … More zymatic digestion for chitin and chitosan biodegradation in bone tissue. DDAC 100 should be a suitable biomaterial for bone surgery and bone regeneration therapy.
The other study was undertaken to evaluate the applicability of chitosan monomer (D-glucosamine hydrochloride) as a pulp capping medicament. Both in vitro and in vivo experiments were carried out to study the cell metabolism and wound healing mechanisms following the application of chitomonosaccharide. After 3 days of osteoblast culture, alkaline phosphatase (ALP) activity significantly increased in the chitosan group. Reverse transcription polymerase chain reaction analysis revealed that chitosan induced an increase in the expression of ALP mRNA after 3 days and bone morphogenetic protein-2 mRNA after 7 days of osteoblast incubation. Inflammatory cytokine, interleukin (IL)-8, synthesis in fibroblasts was strongly suppressed in the medium supplemented with chitosan monomer. Histopathological effects were evaluated in rat experiments. After 1 day, inflammatory cell infiltrations were observed to be weak when compared with the application of chitosan polymer. After 3 days, a remarkable proliferation of fibroblasts was seen near the applied chitosan monomer. The inflammatory cell infiltration had almost completely disappeared. After 5 days, the fibroblastic proliferation progressed, and some odontoblastic cells appeared at the periphery of the proliferated fibroblasts. These findings indicate that the present study is the first report that chitosan monomer acts as a biocompatibly stable medicament even at the initial stage of wound healing in comparison with the application of chitosan polymer. Less

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (10 results)

All 2007 2006 2005

All Journal Article (10 results)

  • [Journal Article] Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer2007

    • Author(s)
      Tomoko Ganno
    • Journal Title

      Jornal of Biomedical Materials Reserch A (In press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Chitosan monomer accelerates alkaline phosphatase activity on-human osteoblastic cells under hypofunctional conditions2007

    • Author(s)
      Shizuka Yamada
    • Journal Title

      Jornal of Biomedical Materials Reserch A (In press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer2007

    • Author(s)
      Tomoko Ganno, Shizuka Yamada, Naoko Ohara, Tsunenori Matsunaga, Kajiro Yanagiguchi, Takeshi Ikeda, Hidetaka Ishizaki, Yoshihiko Hayashi
    • Journal Title

      Jornal of Biomedical Materials Reserch (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Chitosan monomer accelerates alkaline phosphatase activity on human osteoblastic cells under hypofunctional conditions2007

    • Author(s)
      Shizuka Yamada, Tomoko Ganno, Naoko Ohara, Yoshihiko Hayashi
    • Journal Title

      Jornal of Biomedical Materials Reserch (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Early gene expression analyzed by cDNA microarray and real-time PCR in osteoblasts cultured with chitosan monomer2007

    • Author(s)
      Tomoko Ganno
    • Journal Title

      Jornal of Biomedical Materials Reserch A 80(5)(In press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Chitosan monomer promotes tissue regeneration on dental pulp wounds2006

    • Author(s)
      Tsunenori Matsunaga
    • Journal Title

      Jornal of Biomedical Materials Reserch A 76A

      Pages: 711-720

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Chitosan monomer promotes tissue regeneration on dental pulp wounds.2006

    • Author(s)
      T.Matsunaga, K.Yanagiguchi, S.Yamada, N.Ohara, T Ikeda, Y.Hayashi
    • Journal Title

      Jornal of Biomedical Materials Reserch 76A

      Pages: 711-720

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Chitosan monomer promotes tissue regeneration on dental pulp wounds2006

    • Author(s)
      Tsunenori Matsunaga
    • Journal Title

      Journal of Biomedical Materials Research 76A

      Pages: 711-720

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Immunohistochemical and electron microscopic study of the biodegradation processes of chitin and chitosan implanted in rat alveolar bone.2005

    • Author(s)
      Takeshi Ikeda
    • Journal Title

      Oral Medicine & Pathology 10(4)

      Pages: 131-138

    • NAID

      130004509478

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Immunohistochemical and electron microscopic study of the biodegradation processes of chitin and chitosan implanted in rat alveolar bone.2005

    • Author(s)
      T.Ikeda, K.Yanagiguchi, T.Matsunaga, S.Yamada, N Ohara, T.Ganno, Y.Hayashi
    • Journal Title

      Oral Medicine & Pathology 10(4)

      Pages: 131-138

    • NAID

      130004509478

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary

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Published: 2005-04-01   Modified: 2016-04-21  

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