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2010 Fiscal Year Final Research Report

Investigation for establishment of periodontal regenerative therapy using stem cells in cultured periodontal ligament cells

Research Project

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Project/Area Number 20592436
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

KOBAYASHI Makoto  昭和大学, 歯学部, 講師 (80186767)

Co-Investigator(Kenkyū-buntansha) USUI Michihiko   (10453630)
TAKADA Takatora   (20384323)
Project Period (FY) 2008 – 2010
Keywordsヒト歯根膜 / 間葉系幹細胞 / 歯周組織再生 / Side Population(SP)細胞 / 骨分化制御 / BMPアンタゴニスト / twist / biglycan / decorin
Research Abstract

In 2008 : The various types of stem cells exist in side population(SP) effused strongly Hoechst 33342 dye. The SP cells in bone marrow express cell surface markers of human stem cells(HSCs) and are highly enriched for HSC activity. In this study, we clarified the characteristics of SP cells isolated from human periodontal ligament(HPDL) cells. Unfortunately, the present data concluded that HPDL cells contain SP cells, whereas the SP cells are not enriched for mesenchymal stem cell(MSC)-like properties.
In 2009, Twist and bone morphogenetic protein(BMP) antagonists may play an important role in maintenance of constancy of PDL, by which negatively regulate osteoblastic differentiation of cells existing in PDL. In this study, changes of these molecules expression by inducing osteoblastic differentiation were assessed in HPDL cells. We further investigated regulatory effects of BMP antagonists on osteoblastic differentiation in the cells. The present results suggest that under physiological … More condition, twist and BMP antagonists ; gremlin and follistatin, which are constitutively expressed in HPL cells, negatively regulate osteoblastic differentiation of the cells. Further, the induction of osteoblastic differentiation in HPL cells simultaneously enhances twist expression, while it suppresses expression of gremlin and follistatin, as the result, the osteoblastic differentiation may be modulated.
PDL contains fibroblastic cells, which may participate in formation and maintenance of PDL fibers, and osteoblastic cells with a potential to generate alveolar bone and cementum, however, the specific markers for recognition and isolation of each cell type have not been clarified. The purpose of this study was to isolate alkaline phosphatase(ALP)-positive and-negative subpopulations from HPDL cells using flowcytometry and to identify functional difference between the 2 subsets in terms of mitogenic activity, osteogenic potential and expression of decorin as well as biglycan. The present results suggest that ALP-positive HPDL cells have osteoblast-like phenotype, while ALP-negative HPDL cells are fibroblastic cell population with weak osteogenic potential. Further, biglycan may positively regulate osteoblastic differentiation in ALP-positive HPDL cells, while decorin, whose expression is increased by osteogenic induction, may negatively regulate mineralization in both subpopulations.
In 2010, to investigate characteristics of MSC existed in PDL, CD271+/CD90+cells, which were isolated from HPDL by FACS, were single cell cultured, and the clones with MSC-like properties were collected. Therefore, CD271+/CD90+may be a marker to collect MSC from HPDL. Less

  • Research Products

    (6 results)

All 2009 2008

All Journal Article (3 results) (of which Peer Reviewed: 3 results) Presentation (3 results)

  • [Journal Article] ヒト歯根膜細胞中に存在するアルカリフォスファターゼ陽性細胞と陰性細胞の特徴2009

    • Author(s)
      鶴見亜有子, 小林誠, 村山怜一郎ら, (2番目)
    • Journal Title

      Dental Med Res

      Volume: 29(1) Pages: 28-39

    • Peer Reviewed
  • [Journal Article] 培養ヒト歯根膜細胞におけるTwistと各種BMP antagonistの発現制御2008

    • Author(s)
      森高俊一郎, 小林誠, 三井将ら, (2番目)
    • Journal Title

      Dental Med Res

      Volume: 28(2) Pages: 67-76

    • Peer Reviewed
  • [Journal Article] 培養ヒト歯根膜細胞における各種腱・靱帯マーカー遺伝子の発現制御2008

    • Author(s)
      三井将, 小林誠, 森高俊一郎ら, (2番目)
    • Journal Title

      Dental Med Res

      Volume: 28(3) Pages: 137-149

    • Peer Reviewed
  • [Presentation] ヒト歯根膜細胞中のアルカリフォスファターゼ陰性細胞の特徴2008

    • Author(s)
      鶴見亜有子, 小林誠, 奥津朋彦ら
    • Organizer
      第129回日本歯科保存学会秋期学術大会
    • Place of Presentation
      富山
    • Year and Date
      20081100
  • [Presentation] Characteristics of side cell population cells existing in periodontal ligament cells2008

    • Author(s)
      Koide Y, Kobayashi M, et al.
    • Organizer
      The 86th General Session & Exhibition of the IADR
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      20080600
  • [Presentation] 培養ヒト歯根膜細胞中に存在するSide Population細胞の特徴2008

    • Author(s)
      小出容子, 小林誠ら
    • Organizer
      第7回日本再生医療学会総会
    • Place of Presentation
      名古屋
    • Year and Date
      20080300

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Published: 2013-07-31  

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