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Celluar Biological Analysis on the Mechanism of Mineralization of Human Bone Tissue and its Application for Reconstruction of Human Tissue in Vitro

Research Project

Project/Area Number 62440077
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 外科・放射線系歯学
Research InstitutionTokushima University

Principal Investigator

NAGAYAMA Masaru  Tokushima Univ., School of Dentistry, Professor, 歯学部, 教授 (30022867)

Co-Investigator(Kenkyū-buntansha) GOTOH Yuji  Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (60225670)
TAKISHITA Yukio  Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (30226964)
HAYASHI Eiji  Tokushima Univ., Univ. Dental Hospital, Research Assoc., 歯学部・附属病院, 助手 (50173000)
YASUDA Katsuhiro  Tokushima Univ., School of Dentistry, Research Assoc., 歯学部, 助手 (00174508)
HIRAIWA Kiyotaka  Tokushima Univ., Univ. Dental Hospital, Assist. prof., 歯学部・附属病院, 講師 (30173214)
荘田 彰  徳島大学, 歯学部附属病院, 助手 (60196996)
Project Period (FY) 1987 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥17,200,000 (Direct Cost: ¥17,200,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1988: ¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 1987: ¥6,900,000 (Direct Cost: ¥6,900,000)
Keywordshuman osteoblastic cells / collagen gel / platelet-derived growth factor / bone marrow stromal cell / in vitro mineralization / chondrocytes / retinoic acid / endochondral ossification / ヒト骨組織 / ヒト骨芽細胞 / 石灰化 / PDGF / プロテインキナ-ゼC / カルシウム / オステオカルシン / アルカリ性ホスファタ-ゼ / ヒト骨組識 / PTH / 骨吸収因子 / 骨代謝 / クローニング / コラーゲンゲル / 精製
Research Abstract

Osteoblastic cells were isolated from human maxilla by embedding the bone pieces in collagen gel. As a result of the culture and characterization of the isolated cells, the cells responded to PTH and PGE_2, secreted osteocalcin and formed mineralized area in monolayer culture. When we investigated the effect of several factors on differentiation and proliferation of the cells, platelet-derived growth factor contained in the bone matrix tended to stimulate proliferation of the cells.
Bone marrow tissue is reported to contain cells which can differentiate to osteoblast and chondroblast. From these facts, we cultured bone marrow stromal cells and elucidated that the tissue has osteogenic cells, which formed mineralized nodules in vitro. The nodules were analyzed histologically and immunohistochemically. Among the cells, it is confirmed to exist the cells which could mineralize without beta-glycerophosphate.
As chondrocytes have an important role in bone formation, we have also studied the f … More ollowing research. We obtained terminally differentiated chondrocytes in monolayer culture. The cells tended to be stimulated proliferation by retinoic acid. The results suggested that the mechanism of proliferation by retinoic acid was not mediated by epidermal growth factor but by protein kinase C. The retinoic acid binding capacity was detected in both the cytosol and nuclear of the cultured chondrocytes. These binding capacity was supposed to be involved in the mechanism of retinoic acid.
Furthermore, in order to obtain more insight into the physiologic mechanism of endochondral ossification, changes occurring in the mandibular cartilage of growing rats fed on a low-calcium diet were investigated. The results indicated that calcium-deficiency and subsequent metabolic disorder in growing rats induced inhibition of cell proliferation in the proliferative cell zone, inhibition of maturation of chondrocytes, and inhibition of cartilage resorption due to poor calcification of the extracellular matrix, so that the mechanism of normal endocondral ossification might be inhibited and enibited and endochondral growth of mandibular condyle by suppressed. Less

Report

(5 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • 1988 Annual Research Report
  • 1987 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] 松本 文博: "低カルシウム食飼育成長期ラットにおける下顎頭軟骨の組織学的研究" 四国歯学会誌. 3. 161-177 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yuji Goton: "In vitro mineralization of osteoblastic cells derivered from human bone" Bone and Mineral. 8. 239-250 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yukio Takishita: "Effect of Retinoic Acid on Proliferation and Differentiation of Cultured Chondrocytes in Terminal Differentiation" the Journal of Biochemistry. 107. 592-596 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Fumihiro Matsumoto: "Light and Electron Microscopic Observation on the mandiblular Condylar Cartilage in Growing Rats on Lowーcalcium Diet" Acta Anatomica,in prees.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kazuhito Satomura: "Mineralized Nodule Formation in Rat Bone Marrow Cell Culture without βーGlycerophosphate" Bone and Mineral.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kazuhiro Satomura: "Ultrastructure of Mineralized Nodule Formed in Rat Bone Marrow Stromal Cell Culture"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kenichiro Tamai: "Role of protein Kinase C on Retinoic acidーinduced Growth Promotion of Cultured Chondrocytes in Terminal Differentiation"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kenichiro Tamai: "Phosphorylation of Cultured Chondrocytes induced by Retinoic Acid"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hiroaki Nakanishi: "Effect of Plateletーderived Growth Factor on Proliferation of Cultured Human Osteoblastic Cells"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Fumihiro MATSUMOTO: "Histological Observations on the Mandibular Condylar Cartilage in Growing Rats on a Low-calcium Diet" Shikokushigakkaishi(in Japanese).3(1). 161-177 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yuji GOTOH kiyotaka HIRAIWA, Masaru NAGAYAMA: "In vitro mineralization of osteoblastic cells derived from human bone" Bone and Mineral. 8(3). 239-250 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yukio TAKISHITA, Kiyotaka HIRAIWA, Masaru NAGAYAMA: "Effect of Retinoic Acid on Proliferation and Differentiation of Cultured Chondrocytes in Terminal Differentiation" Journal of Biochemistry. 107(4). 592-596 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Fumihiro MATSUMOTO, Youji MIYAMOTO, Masaru NAGAYAMA: "Light and Electron Microscopic Observations on the Mandibular Condylar Cartilage in Growing Rats on a Low-calcium Diet" Acta Anatomica,.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kazuhito SATOMURA, Kiyataka HIRAIWA, Masaru NAGAYAMA: "Mineralized Nodule Formation in Rat Bone Marrow Cell Culture without beta-Glycerophosphate" Bone and Mineral,.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kazuhiro SATOMURA, Masaru NAGAYAMA: "Ultrastructure of Mineralized Nodule Formed in Rat Bone Marrow Stromal Cell Culture"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kenichiro TAMAI, Yukio TAKISHITA, Kiyotaka HIRAIWA, Masaru NAGAYAMA: "Role of Promotion of Kinase C on Retinoic acid-induced Growth Promotion of Cultured Chondrocytes in Terminal Differentiation"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kenichiro TAMAI, Yukio TAKISHITA, Kiyotaka HIRAIWA, Masaru NAGAYAMA: "Phosphorylation of Cultured Chondrocytes induced by Retinoic Acid"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hiroaki NAKANISHI, Yuji GOTOH, Kiyotaka HIRAIWA, Masaru NAGAYAMA: "Effect of Platelet-derived Growth Factor on Proliferation of Cultured Human Osteoblastic Cells"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yukio Takishita: "Effect of Retinoic Acid on Proliferation and Differentiation of Cultured Chondrocytes in Terminal Differentiation" the Journal of Biochemistry. 107. 592-596 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kazuhito Satomura: "Mineralized Nodule Formation in Rat Bone Marrow Cell Culture without βーGlycerophosphate" Bone and Mineral.

    • Related Report
      1990 Annual Research Report
  • [Publications] Fumihiro Matsumoto: "Light and Electron Microscopic Observations on the mandibular Condylar Cartilage in Growing Rats on a Lowーcalcium Diet" Acta Anatomica.

    • Related Report
      1990 Annual Research Report
  • [Publications] Kenichiro Tamai: "Role of Protein Kinase C on Retinoic acidーinduced Growth Promotion of Cultured Chondrocytes in Terminal Differentiation"

    • Related Report
      1990 Annual Research Report
  • [Publications] Kenichiro Tamai: "Phosphorylation of Cultured Chondrocytes induced by Retinoic Acid"

    • Related Report
      1990 Annual Research Report
  • [Publications] Hiroaki Nakanishi: "Effect of Plateletーderived Growth Factor on Proliferation of Cultured Human Osteoblastic Cells"

    • Related Report
      1990 Annual Research Report
  • [Publications] Yuji Gotoh: "In vitro mineralization of osteoblastic cells derived from human bone" Bone and Mineral. 8. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yukio Takishita: "Effect of Retionic Acid on Proliferation and Differentiation of Cultured Chondrocytes in Terminal Differentiation" the Journal of Biochemistry. 107. (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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