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Study on The Dentinal Native Non-collagenous Protease

Research Project

Project/Area Number 03670904
Research Category

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

Allocation TypeSingle-year Grants
Research Field Conservative dentistry
Research InstitutionKanagawa Dental College

Principal Investigator

TERANAKA Toshio  Kanagawa Dental Coll.Dept.of Restorative Dent.Assoc.Prof., 歯学部, 助教授 (60104460)

Co-Investigator(Kenkyū-buntansha) SAITO Masahiro  Kanagawa Dental Coll.Dept.of Restorative Dent.Instructor, 歯学部, 助手 (40215562)
SATOYOSHI Masanori  Kanagawa Dental Coll.Dept.of Restorative Dent.Instructor, 歯学部, 助手 (90257303)
MUKAI Yoshiharu  Kanagawa Dental Coll.Dept.of Restorative Dent.Instructor, 歯学部, 助手 (40247317)
川野 賢哉  神奈川歯科大学, 歯学部, 助手 (60214665)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsDentin / Odontoblast / Dentin bonding / Dentin Extract / Protease / Metalloprotease / Stromelysine / Proteglycan / 石灰化 / リン蛋白 / エンザイモグラム / メタロプロテア-ゼ
Research Abstract

The objectives of this research project were 1) to extract the non-collagenous dentin protein hydrolyzable protease from dentin, 2) to study the hydrolytic ability of the protease against the dentin proteins and proteoglycans, 3) to establish the mineralizing cell culture and 4) to investigate the influence of different dentin adhering mechanisms on shear and fatigue bond strength.
The results obtained were as follows :
1.Several proteases were extracted from human and bovine dentin. The approximate molecular weight of 59KDa (59K P-ase) showed the major enzymatic activity and its optimum pH was 9.59K P-ase was inhibited by EDTA and TIMP, and cleaved osteopontin. It was suggested that 59K P-ase is a phophoprotein protease and play an important role in both dentin matrix development and degradation process.
2.Both the EDTA and guanidium chloride extracted dentin proteoglycans were degraded into small fragments by 59K P-ase. It was conceivable that the 59K P-ase was one of metalloproteases b … More ut not a collagenase, and play an important role in the process of mineralization and maturation of dentin.
3.Several dentin proteoglycans were obtained, ther molecular weights were 150-180KD, 300KD, 130-150KD and 180-210KD on SDS/PAGE.59K P-ase degraded all of these proteoglycans. E-ext proteoglycan has several core proteins which were stained blue with Stains-all and these core proteins were also cleaved by 59K P-ase. These results demonstrate that the 59K P-ase has a stromelysin-like activities.
4.Odontoblast derived from bovine incisors was cultured on a reconstituted gel of basement membrane components, produced a dentin-specific protein phosphopholyn and grew mineralized nodules. The highest concentration of the enzyme related in the matrix was observed between day 22 and day 36 coinciding with the initiation of mineralization. The described sequence of developmental expression of proteins in this mineralizing dentin cell culture is very similar to that in bone which suggests common mechanisms of matrix mineralization in bone and dentin.
The shear and fatigue shear bond strength of a third generational dentin bonding agent (KB-110) was compared with a conventional one. KB-110 showed significantly higher shear and fatigue bond strength than conventional, though its fatigue strength decreased rapidly as the storage period prolonged. These results suggest that the fatigue strength reflects more accurately the retentive properties of the dentin matrix than the shear bond strength. Less

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] 斎藤 正寛: "59KDプロテアーゼの基質となる象牙質中のプロテオグリカンに関する研究" 歯基礎誌. 36(投稿中). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 里吉 正徳: "Mineralization Process in The Bovine Odontoblast Culture" Bone and Mineral Research. (投稿中). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 里吉 正徳: "培養ウシ象牙芽細胞の分化に関する研究" 日歯保誌. 36. 495-506 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 向井 義晴: "象牙質中のプロテオグリカン分解酵素に関する研究" 日歯保誌. 35. 422-432 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 里吉 正徳: "エンザイモグラムにおけるProteaseの挙動" 歯基礎誌. 35. 186-196 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 向井 義晴: "ヒト象牙質中に存在するリン蛋白分解酵素に関する研究" 歯基礎誌. 33. 61-69 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Saito, M., Mukai, Y., Tomiyama, K., Teranaka, T., Iwamoto, T., Kubota, T., Sato, S.& Suzuki, Y.: "A study on dentin proteoglycans as substrates for dentin 59KD protease." Jpn J Oral Biol. 36-2 in press. (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Satoyosi, M., Koizumi, T., Teranaka, T., Iwamoto, T., Kakai, Y., Kawase, T., Saito, S.& Mikuni-Takagaki, Y.: "Mineralization processes in the bovine odontoblast culture." Bone and Mineral Research. in press. (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Mitsui, K.& Teranaka, T.: "Influence of fatigue on bond strength of dentin bonding system" J J Consev Dent.37-3 in press. (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Satoyoshi, M., & Mikuni-Takagaki, Y.: "Differntiation of odontoblastic cells in culture" J J Consev Dent.36-2. 495-506 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Satoyoshi, M., T., Teranaka, T., Iwamoto, T., Imai, K., Kawase, T.& Saito, S.: "Electrophoretic behavior of protease on enzymogram : Estimation of the molecular weight." Jpn J Oral Biol.35-2. 186-196 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Mitsui, K., Matsuzawa, S., Koizui, T., Hanaoka, K.Teranaka, T.& Iwamoto, T.: "Adhesive characteristics of light-settings glass ionomer cements." Kanagawa Shigaku. 27-4. 569-576 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Akutagawa, T.: "A study on the apical marginal adaptaion of composite restoration" J J Consev Dent.36-1. 1-12 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Mukai, Y.: "Study of proteoglycan degrading metalloprotease in dentin." J J Consev Dent.35-2. 422-432 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Mukai, Y., Saito, M., Teranaka, T., Iwamoto, T.Kubota, T., Sato, S.& Suzuki, Y.: "59KDa phosphoprotein proteas from human dentin." Jpn J Oral Biol.33-1. 61-69 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 里吉,正徳: "培養ウシ象牙芽細胞の分化に関する研究" 日歯保誌. 36. 495-506 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 斎藤,正寛: "ウシ象牙質抽出分画によるヒト歯根膜線維芽細胞の接着・伸展について" 日歯保誌. 36. 3 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 斎藤,正寛: "Cell Attachment and Spreading Factors of Bovine Dentine Extract" J.Dent.Res.73. 148 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 里吉,正徳: "Posttranslational Processing of Dentin Phosphoprotein(s) in Odontoblast-loke Cells" J.Dent.Res.73. 147 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 向井 義晴: "象牙質中のプロテオグリカン分解酵素に関する研究" 日歯保誌. 35. 422-432 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 里吉 正徳: "エンザイモグラムにおけるproteaseの挙動" 日基礎誌. 34. 61-69 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 斎藤 正寛: "59KDプロテアーゼの基質になる象牙質中のプロテオグリカンについて" 日骨代謝誌. 10. 280- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 里吉 正徳: "培養ウシ象牙芽細胞における象牙質リン酸化酵素に対する研究" 日基礎質. 34. 137- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 里吉 正徳: "培養ウシ象牙芽細胞の分化と基質石灰化" 日歯保誌. 35. 17- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 富山 潔: "象牙質プロテオグリカンの抽出とその特性に関する研究" 日歯保誌. 35. 18- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 向井 義晴: "ヒト象牙質中に存在するリン蛋白分解酵素に関する研究" 歯基礎誌. 33. 61-69 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 向井 義晴: "象牙質中のプロテオグリカン分解酵素に関する研究" 日歯保誌. 35. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Saito,Masahiro: "59KD protease in dentin.Isolation and characterization from bovine dentin" J.dent.Res.70. 310 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mukai,Yoshiharu: "Isolation and partial characterization of Mr 59000 stromelysineーlike protease in dentin" Connective Tissue Res.27. 113 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 里吉 正徳: "Enzymogramにおけるプロテア-ゼの挙動" 日歯周誌. 33. 87 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 向井 義晴: "象牙質中のProteaseに関する研究(III)" 日歯保誌. 34. 116 (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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