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1989 Fiscal Year Final Research Report Summary

Role of the osteoclast and osteoblast during the bone resorption

Research Project

Project/Area Number 63480406
Research Category

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

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionShowa University

Principal Investigator

YOSHIKI Shusaku  Showa University, School of Dentistry, Professor, 歯学部, 教授 (30085740)

Co-Investigator(Kenkyū-buntansha) TAKAHASHI Naoyuki  Showa University, School of Dentistry, Assistant Professor, 歯学部, 講師 (90119222)
YAMAGUCHI Akira  Showa University, School of Dentistry, Associate Professor, 歯学部, 助教授 (00142430)
Project Period (FY) 1988 – 1989
Keywordsosteoblast / osteoclast / bone resorption / collagenase / PTH / 1,25(OH)_2D_3 / 副甲状腺ホルモン
Research Abstract

The object of this Project is to investigate each function of the osteoclast and osteoblast during bone resorption process. We applied in vitro culture systems for studying the functions. The results obtained from this project are as follows.
(1)Function o-f the osteoblast a)Immunoreactive-collagenase was detected in all of the osteoblastic cell lines tested (5 clonal rat osteoblastic cell lines established by ourselves and a rat osteosarcoma cell line, UMR-106) using a rabbit anti-rat collagenase antibody by an indirect immunoperoxidase technique. b)Tissue plasminogen activator, which is a stimulator of latent form-collagenase, was also detected in all of the osteoblastic cell lines tested by an immunohistochemical technique. c) Collagenase activity was measured using as assay kit [collagenokit]. This kit contains fluorescence- conjugated collagen as substrate for collagenase. We found no significant increase of collagenase activity in the osteoblastic cell lines between basal level an … More d stimulated conditions with PTH or 1,25(OH)_2D_3. These results suggested to test the more sensitive assay system, including [14C] labeled collagen 'for the detection of the collagenase activity in the osteoblastic cells. d) We developed new assay system for the measurement of bone degradation;osteoblastic cells were cultured on the 13H]-proline labeled calvaria with or without 1,25(OH)_2D_3, and the level of 13H) was determined in the medium and bone after 5 days culture. As a result, 1,25(OH)_2D_3 stimulated the degradation of collagenous matrix by the osteoblastic cells.
a)Immunoreactive-collagenase was detected in all of the osteoblastic cell lines tested (5 clonal rat osteoblastic cell lines established by ourselves and a rat osteosarcoma cell line, UMR-106) using a rabbit anti-rat collagenase antibody by an indirect immunoperoxidase technique. b)Tissue plasminogen activator, which is a stimulator of latent form-collagenase, was also detected in all of the osteoblastic cell lines tested by an immunohistochemical technique. c) Collagenase activity was measured using as assay kit [collagenokit]. This kit contains fluorescence- conjugated collagen as substrate for collagenase. We found no significant increase of collagenase activity in the osteoblastic cell lines between basal level and stimulated conditions with PTH or 1,25(OH)_2D_3. These results suggested to test the more sensitive assay system, including [^<14>C] labeled collaben, for the detection of the collagenase activity in the osteoblastic cells. d) We developed new assay system for the measurement of bone degradation;osteoblastic cells were cultured on the [3H]-proline labeled calvaria with or without 1,25(OH)_2D_3, and the level of [3H] was determined in the medium and bone after 5 days culture. As a result, 1,25(OH)_2D_3 stimulated the degradation of collagenous matrix by the osteoblastic cells.
(2)Study for the osteoclast formation
a) We developed an in vitro assay system for the osteoclast formation from mouse bone marrow cells. b)Using this system, we found that 1,25(OH)_2D_3, PTH, IL-1, PGE_2 stimulate the osteoclast formation, and calcitonin and interferon-gamma inhibit its formation. c) We demonstrated that mouse spleen cells have a potential to develop the osteoclast. In this process, spleen cells required the presence of the osteoblastic cells, 1,25(OH)_2D_3 and dexamethasone. Less

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Udagawa,N.,Takahashi,N.,Akatsu T.,Sasaki,T.,Yamaguchi,A.,Kodama,H.,Martin,T.J.,Suda,T.: "The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclastlike cell deffentiation in cocultures with mouse spleen cells." Endocrinology. 125. 1805-1813

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 吉木周作: "脱灰切片による硬組織内物質の組織形態的観察法に関する一連の研究" 昭和歯学会雑誌. 8. 247-255 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山口朗,吉木周作: "骨芽細胞の形態と機能の多様性" 臨床化学. 25. 241-248 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山口朗,吉木周作: "骨のコラゲナ-ゼ" 骨代謝誌. 7. 50-57 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山口朗,池田通,吉木周作: "骨芽細胞の培養" 組織培養. 15. 155-159 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi,A.,Kahn,A.J.: "Clonal osteogenic cell lines exeress mygenic and adivocytic developmental potential" (投稿中).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamaguchi, A., Yoshiki, S.: "Diversity of phenotype and function in osteoblast." J. Clin. Sci. 25, 241-248, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, A., Yoshiki, S.: "Bone collagenase." Jap. Soc. Bone Mineral Res. 7.50-57, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, A., Ikeda, T., Yoshiki, S.: "Cell culture of osteoblastic cells." The Tissue Culture 15, 155-159, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshiki, S.: "Histochemical observation of mineralized matrix on decalcified sections." J. Showa Univ. Dent. Soc. 8, 247-255, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, N., Yamana, H., Yoshiki, S., Roodman, G.D., Mundy, G.R., Jones, S.J., Boyde, A., Suda, T.: "Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow" Endocrinology; 122, 1373-1382, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, N., Akatsu, T., Udagawa, N., Sasaki, T., Yamaguchi, A., Moseley, J. M., Martin, T. J., and Suda, T.: "Osteoblastic cells are involved on osteoclast formation." Endocrinology; 123, 2600-2602, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Udagawa, N., Takahashi, N., Akatsu, T., Sasaki, T., Yamaguchi, A., Kodama, H., Martin, T.J., Suda, T.: "The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclast-like cell differentiation in cocultures with mouse spleen cells." Endocrinology; 125, 1805-1813, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaguchi, A., Kahn, A.J.: "Clonal osteogenic cell lines express myogenic and adipocytic developmental potential." submitted.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-26  

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