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A novel inhibitor in bone formation from periodontopathic bacteria

Research Project

Project/Area Number 07557137
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 矯正・小児・社会系歯学
Research InstitutionKyushu Dental College

Principal Investigator

TAKEHARA Tadamichi  Kyusyu Dental College Prof., 歯学部, 教授 (00038879)

Co-Investigator(Kenkyū-buntansha) MATSUO Noboru  Institute of Kao, 主任研究員
ANSAI Toshihiro  Kyusyu Dent.Coll.Associate Prof., 歯学部, 助教授 (80244789)
HANEJI Tatsuji  Kyusyu Dent.Coll.Associate Prof., 歯学部, 助教授 (50156379)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1995: ¥2,800,000 (Direct Cost: ¥2,800,000)
KeywordsOral bacteria / osteoblast / Prevotella intermedia / Actinobacillus / アルカリホスファターゼ / 骨芽細胞 / アルカリ性フォスファターゼ / 歯周病菌 / Prevotella intermedia / Actinobacillus actinomycetemcomitans
Research Abstract

In order to study the effects of sonicated extracts from Prevotella intermedia. Actinobacillus actinomycetemcomitans, Porphyomonas gingivalis, and other oral-related bacteria, as well as Escherichia coli on bone formation, clone MC3T3-E_1 cells, which have retained osteoblastic activity, were cultured with various bacterial extracts.
The addition of the sonicated extracts from Prevotella intermeedia and Actinobacillus actinomycetemcomitans decreased the alkaline phosphatase activity in a dosedependent fashion over the concentration range of 1-1000 ng ml^<-1> compared with the control. By contrast, the sonicated extracts from other oral bacteria including Prophyromonas gingivalis, Capnocytophaga ochracea, Streptococcus milleri and Streptococcus sanguis, and Escherichia coli did not decrease the alkaline phosphatase activity even in the presence of 100 ng ml^<-1> protein.
The addition of prevotella intermedia and Actinobacillus actinomycetemcomitans extracts that had been treated with heat and trypsin to the cell cultures also inhibited alkaline phosphatase activity in the cells, suggesting that inhibitory factors are not proteinaceous. Polymyxin B did not change the alkaline phosphatase activity in the cells treated with the extracts from Prevotella intermedia and Actinobacillus actinomycetemcomitans, suggesting that the inhibitory activity of the extracts is not lipopolysaccharide.
The inhibitory effect of both extracts was observed in the molecular mass over 290 kDa eluted from Sephadex G-200 column. The inhibitory substances of Prevotella intermedia were partially purified and showed broad band with estemated molecular weight of 170-190 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
These results indicate that Prevotella intermedia and Actinobacillus actinomycetemcomitans may play an important role in inhibiting bone formation as well as in stimulating bone resorption.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Shirakawa, S.et al.: "Immunohistochemical and immunoblotting identification..." FEBS Lett.393. 57-59 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yoshioka, C et al.: "Identification of Fas antigen in human gingiva" J.Dent.Res.75. 1353-1357 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Murata, T.et al.: "Extracts of Prevotella intermedia and Actinobacillus..." Oral Diseases. 3. 106-112 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morimoto, Y.et al.: "The protein phosphatase inhibitors..." Exp.Cell Res.230. 181-186 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morimoto, Y.et al.: "Extracts of Actinobacillus actinomycetemcomitans..." J.Dent.Res.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ansai, T.et al.: "Purification and characterization of alkaline phosphatase..." FEBS Lett.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shirakawa, S., Mochizuki, H., Takehara, T., Kobayashi, S., Shima, H., Nagano, M.and Haneji, T.: "Immunohistochemical and immunoblotting identification of protein phosphatase lgammal in rat salivary glands." FEBS letters. 393. 57-59 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yoshioka, C., Muraki, Y., Fukuda, J., Haneji, T.and Kobayashi, N.: "Identification of Fas antigen in human gingiva." Journal of Dental Research. 75. 1353-1357 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Murata, T., Ansai, T., Takehara, T., Kobayashi, S.and Haneji, T.: "Extracts of Prevotella intermedia and Actinobacillus actinomycetemcomitans inhibit alkaline phoshatase activity in osteoblastic cells in vitro." Oral Diseases. 3. 106-112 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morimoto, Y., Ohba, T., Kobayashi, S.and Haneji, T.: "The protein phosphatase inhibitors okadic acid and calyculin A induce apoptosis in human osteoblastic cells." Experimental Cell Research. 230. 181-186 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morimoto, Y., Morimoto, H., Ohba, T., Kobayashi, S.and Haneji, T.: "Extracts of Actinobacillus actinomycete-comitans induce apoptotic cell death in human osteoblastic MG63 cells." Journal of Dental Research. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ansai, T., Awano, S., Chen, X., Fuchi, T., Arimoto, T., Akifusa, S.and Takehara.T.: "Purification and characterization of alkaline phosphatase containing phosphotyrosyl phosphatase activity from the bacterium Prevotella intermedia." FEBS letters. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shirakawa,S et al.: "Immunohistochemical and immunoblotting identification of protein phosphatase 1 1 in salivary glands" FEBS letters. 393. 57-59 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Murata,T.et al.: "Extracts of Prevotella intermedea and Actinobacillus..." Oral Diseases. 3. 106-112 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yoshioka,C.et al.: "Identification of Fas antigen in human gingiva" J.Dent.Res.75(6). 1353-1357 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Morimoto,Y.et al.: "The protein phosphatase inhibitors Okadaic acid and Calyculin A.." Experimental Cell Research. 230(2). 181-186 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Morimoto,Y.et al.: "Extracts of Actinobacillus actinomycetemcomitans induce..." J.Dent.Res.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Shirakawa,et al.: "Transblot Identification of Avidin-Interacting Proteins in Rat Salivary Glands" Biochemistry and Molecular Biology International. 40(1). 67-72 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Shirakawa,et al.: "Immunohistochemical and immunoblotting identification of protein phosphatase 11 in rat salivary glands" FEBS Letters. 393. 57-59 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Murata,et al.: "Extracts of Prevotella intermedia and Actinobacillus actinomyceten comitans inhibit alkaline phosphatase activity in osteoblastic…" Oral Diseases (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] C.Yoshioka,et al.: "Identification of the Fas antigen in human gingiva" Journal of Dental Research. 75(6). 1353-1357 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Muraki,et al.: "Immunohistochemical detection of Fas antigen in oral epithelia" Oral Pathology and Medicine. 26. 57-62 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Morimoto,et al.: "The protein phosphatase inhibitors Okadaic Acid and Calyculin A induce apoptosis in human osteoblastic cells" Experimental Cell Research. 230(2). 181-186 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masahito Kawasaki: "An enzyme-linked immunosorbent assay for measuring antibodies to serotype-specific polysaccharide…" J.Microbiol.Methods. 21. 181-192 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Toshigiro Ansai: "A murC gene in Porphyromonas gingivalis 381" Microbiology. 141. 2047-2052 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takatoshi Murata: "Protein phosphatase inhibitors,Okadaic Acid and Calycuin A,induce alkaline phosphatase activityin…" Biochem.Mol.Biol.Int.36. 365-372 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Chihiro Yoshioka: "Identification of Fas antigen in human gingiva" J.Dent.Res.(in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Takatoshi Murata: "Effect of sonicated extract from Prevotella intermedia Actinobacillus actinomycetemcomitans,…" J.Dent.Res.(in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Yukoh Muraki: "Immunohistochemical detection of Fas antigen in oral epithelia." J.Oral Pathol.Med.(in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Chihiro Yoshioka: "Oral Oncology Vol.IV," Macmillan India, 5 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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