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Histochemical and Ultrastructural Studies on Relationship between Initial Calcification and Lipids

Research Project

Project/Area Number 05671524
Research Category

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

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionKanagawa Dental College

Principal Investigator

TAKAHASHI Tsuneo  Kanagawa D.C., Dentistry, Associate Professor, 歯学部, 助教授 (00130922)

Project Period (FY) 1993 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥900,000 (Direct Cost: ¥900,000)
KeywordsOsteoblast / Intramembranous Occification / Multilamellar bodies / Lipid Droplet / Fatty Acids / Ultrastructure / Calvaria / Mouse / 石灰化 / 多層膜構造 / 膜内石灰化 / 超微構造 / 膜性骨 / タンニン酸 / マラカイト・グリーン / 電子顕微鏡
Research Abstract

This studies aimed at elucidating a biolgical relationship between ossification and lipids in the developmental process of the calvaria of newborn mice at electron microscopic level.
Membranous bone tissues (parietal bone) of newborn mice were employed as the material in both in vivo (1-2 days of age) and in vitro (19 days of gestation) experimental systems. The results are as follows.
(1) Even in the conventional double fixation method with glutaraldehyde and osmium tetroxide, a rapid dehydration procedure using cold acetone solutions after a perfusion fixation method (adjusted at a given pHin the fixative) made it possible to preserve at a higher frequency the substance which exhibits extracellular multilamellar structures.
(2) On the basis of pH experiments for identification and the above findings, it is clear that the main constituents of osteoblastic lipid droplets are not phospholipids or triglycerides. Knowledge obtained strongly suggested that lipid droplets consisted of long-cha … More ined fatty acids.
(3) As another component other than fatty acids contained in lipid droplets, the presence of mucopolysaccharides were also detected. There is also the possibility that thesubstance is a protein complex, but the bond with lipids was not thought tobe a strong bond as seen with the liver and intestine. The morphological relationship between above structures and so-called noncollagenous proteins such as osteocalcin orosteopontin etc. is now being studied.
(4) With respect to extracellular lipids (in a droplet form) of osteoblasts, it is surmised to be composed of unionized (protonated) fatty acids, while ionized fatty acids-soap surrounds the lipid droplets. It should be emphasized that sudanophilic material (lipid-related sustance) described earlier as a substance observed at initial calcifying sites is likely to be unionized (protonated) fatty acids.
(5) Since ionized fatty acids-soap has affinity for calcium, formation of fatty acid-calcium complexes were likely to occur.
In conclusion, these results suggest that the presence of exracellular lipid droplets associated with osteoblasts in this study may play an important role in mineralization as a calcium-tapping mechanism, different from the matrix vesicle-associated calcification. Less

Report

(4 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] 永井宏人: "骨形成過程の骨芽細胞にみられる多層膜構造物の電顕的研究" 神奈川歯学. 29. 333-354 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 高橋常男: "骨形成細胞にみられる多層膜構造物の電顕的研究(第6報)-pH依存症多層膜構造物に対するオスミウム酸の影響-" 日本界面医学会雑誌. 26. 27-36 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tsuneo Takahasi: "Lipid Droplets Associated with Osteoblast by staining with a Mixture of p-Phenylendianine and Imidazole Solution" Bulletine of Kanagawa Dental College. 24. 31-34 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 正木久秀: "骨形成過程の骨芽細胞にみられる多層膜構造物の電顕的研究(第7報)-膜性骨形成とその細胞外脂質について-" 日本界面医学会雑誌. 27. 37-56 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 高橋常男: "骨形成細胞にみられる多層膜構造物の電顕的研究(第7報)-グルタールアルデヒド・マラカイトグリーン固定法によるMLB保存効果" 日本界面医学会雑誌. 27. 136-138 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 上田眞太郎: "肺の発育と老化に関する形態学的研究(第4報)-ウサギとマウスの胎生期における器官と食道の粘膜上皮索の変化ならびに肺表面物質(Sf)との関連性について" 日本界面医学会雑誌. 27. 139-144 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 高橋常男(分担): "走査電子顕微鏡による立体観察" 朝倉書店(印刷中), (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] "An electron microscopic study of multilamellar structures associated with osteoblasts in a mineralization process" 29. 333-354 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tsuneo Takahashi: "An electron microscopic study of multilamellar bodies (MLB) in bone forming cells (Part 6) -Effect of O_5O_4 on appearance of pH dependent MLS -" J.Jpn.Med.Soc.Biol.Interface. 26. 27-36 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tsuneo Takahashi: "Lipid Droplets Associated with Osteoblast by Staining with a Mixture of p-Phenediamine and Imidazole solution" Bull of kanagawa Dent.Col.24. 31-34 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Hisahide Masaki: "An Electron Microscopic Study on Multilamellar Structures in Bone-Forming Cells (Part 7) -The Relationship between Intramembranous Ossification and its Extracellular Lipids-" J.Jpn.Med.Soc.Biol.Interface. 27. 37-56 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tsuneo Takahashi: "An electron microscopic study of multilameller bodies (MLB) in bone forming cells (Part 7) - Preservation of MLB by a glutaraldehyde・malachite green mixed fixative -" J.Jpn.Med.Soc.Biol.Interface. 27. 136-138 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Shintaro Ueda: "Morphological Study on Development and Aging of the Lungs (Part 4) - Relationship between Epithelial Changes of Trachea and Production of Lung Surfactant in Fetal Rabbits nd Mice" J.Jpn.Med.Soc.Biol.Interface. 27. 139-144 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tsuneo Takahashi: Three Dimentional Observations by Scanning Electron Microscope. Asakura Syoten (in print), (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 永井 宏人: "骨形成過程の骨芽細胞にみられる多層膜構造物の電顕的研究" 神奈川歯学. 29. 333-354 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 高橋 常男: "骨形成細胞にみられる多層膜構造物の電顕的研究(第6報)-pH依存性多層膜構造物に対するオスミウム酸の影響-" 日界面医誌. 26. 27-36 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tsuneo Takahasi: "Better visualization of lipid droplets by staining with a mixture of p-phenylenediamine and imidazole solution" Bull of Kanagawa Dent. Col.24. 31-34 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 渡 仲三: "走査型電子顕微鏡による立体組織学" 朝倉書店, 400 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 三宅 実: "唾石表層にみられた多層膜構造物の形態的観察-タンニン酸グルタールアルデヒド混合固定法による-" 日口外誌. 40. 598-603 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yoshinori Jinbu: "Immunohistochemical Study on γ-Carboxyglutamic Acid Containing Protein(osteocalcin,BMP)in Human Dentin and Alveolar Bone" Bull.Kanagawa Dent Col. 22. 03-07 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Tsuneo Takahasi: "pH-dependent multilamellar structures in fetal mouse bone:possible involvement of fatty acids in bone mineralization" Am.J.Physiol.226. 590-600 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Hitoshi Terada: "ENCHONDRAL OSSIFICATION AND VASCULAR ARCHITECTURE ACCOMPANIED WITH GROWTH AND DEVELOPMENT IN RAT MANDIBULAR CONDYLE(Part 2)" Microcirculation annual. 10. 193-194 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Eisei Kono: "VASCULAR ARCHITECTURE OF DOG PULP AFTER PULP CAPPING USING CORROSIVE RESIN CASTS" Microcirculation annual. 10. 195-196 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shuji,SAKAI: "MICROVASCULAR ARCHITECTURE OF THE APICAL PERIODONTIUM BY SCANNING ELECTRON MICROSCOPY OF VASCULAR CORROSION CASTS" Microcirculation annual 1993. 197-200 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tsuneo,Takahashi: "pH-dependent multilamellar structures in fetal mouse bone:possible involvement of fatty acids in bone mineralization" American Journal Physiology. 266(発表予定). (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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