Functional analysis of nitric oxide metabolites in hard tissues

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 18592049
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Showa University
• Principal Investigator: MIYAMOTO Yoichi Showa University, Department of Biochemistry, Associate Professor (20295132)
• Co-Investigator(Kenkyū-buntansha): KAMIJO Ryutaro Showa University, School of Dentistry, Department of Biochemistry, Professor (70233939) ; SUZAWA Ibtsuo Showa University, School of Dentistry, Department of Biochemistry, Associate Professor (60271285) ; TAKAMI Masamichi Showa University, School of Dentistry, Department of Biochemistry, Associate Professor (80307058) ; YAMADA Atsushi Showa University, School of Dentistry, Department of Biochemistry, Associate Professor (50407558) ; AKAIKE Takaaki Kumamoto University, Graduate School of Medical Sciences, Department of Microbiology, Professor (20231798)
• Project Period (FY): 2006 – 2007
• Keywords: chondrocyte / odontoblast / osteoclast / nitric oxide / reactive oxygen / NADPH-oxidase / pH / differentiation

Research Abstract

Nitric oxide (NO) acts as an infra- and intercellular signaling molecule. In addition to the cGMP-dependent signaling, it has been suggested that signals are transmitted by chemical modifications of biological molecules by NO and NO metabolites. In this study, we investigated the roles of NO and its metabolites in the differentiation and function of the cells which constitute hard tissues. First, we established their cell culture models. Namely, odontoblast differentiation, inflammation-induced osteoclast differentiation, and chondrocyte culture as a model of inflammatory joint diseases. We obtained the following findings using these models Expression of inducible type of NO synthase (iNOS) was induced in the pulp after abrasion of teeth without exposure of pulp. Then we studied the effects of NO on the pulp cell growth and differentiation. NO suppressed the proliferation of pulp cells and promoted the odontoblastic differentiation and mineralization, suggesting NO is involved in the f ormation of reparative dentin after tooth abrasion. TNF-α-induced bone resorption in ex-vivo was strongly suppressed in the iNOS-knockout calvaria In addition, osteoclast differentiation in the presence of TNF-α was retarded in the iNOS-knockout cultures NO-dependent formation of nitrated nucleotides was observed in the course of osteockast differentiation in the wild type cells. Finally, we studied the metabolism of extracellular matrix of chondmcytes after IL-1 exposure and found that IL-1 induced and activated the phagocyte-type NADPH-oxidase in the chondrocytes through the activation of TRAF-6 and NF-κB. We had already reported that IL-1 induced the expression of iNOS and formation of peroxynitrite, a reaction product of NO and superoxide in chondrocytes. The present results indicate that one of the sources of superoxide is the phagocyte type NADPH-oxidase. Interestingly, extra- and intracellular pH was decreased in the NADPH-oxidase-dependent manner, which in turn activated hyaluronidase and accelerated the loss of extracellular matrix.

Research Products

• [Journal Article] Characterization of synovial cell clones isolated from rheumatoid arthritis patients : possible involvement of TNF-a in reduction of osteoprotegerin in synovium (2008)
  • Author(s): Zhao, B, Takami, M, Miyamoto, Y, Suzawa, T, Yamada, A, Mochizuki, A, Yasuhara, R, Wang, X, Inoue, T, Namiki, O, Sakamoto, K, Kamijo, R
  • Journal Title: Cytokine 41(1) Pages: 61-70
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] S-Nitrosylation of human variant albumin Liprizzi (R410C) confers potent antibacterial and cytoprotective properties (2007)
  • Author(s): Ishima Y
  • Journal Title: J Pharmacol Exp Ther 320 Pages: 969-977
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Nitric oxide in pulp cell growth,differentiation,and mineralization (2007)
  • Author(s): Yasuhara R
  • Journal Title: J Dent Res 86 Pages: 163-168
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Characterization of synovial cell clones isolated from rheumatoid arthritis patients:possible involvement of TNF-α in reduction of osteoprotegerin in synovium (2007)
  • Author(s): Zhao B
  • Journal Title: Cytokine 41 Pages: 61-70
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Interleukin-4 inhibition of osteoclast differentiation is stronger than that of interleukin-13 and they are equivalent forinduction of osteoprotegerin production from osteoblasts (2007)
  • Author(s): Yamada, A, Takami, M, Kawawa, T, Yasuhara, R, Zhao, B, Mochizuki, A, Miyamoto, Y, Eto, T, Yasuda, H, Namkamichi, Y, Kim, N, Katagiri, T, Suda, T, Kamijo, R
  • Journal Title: Immunology 120(4) Pages: 573-579
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nitric oxide in pulp cell growth, differentiation, and mineralization (2007)
  • Author(s): Yasuhara, R, Suzawa, T, Miyamoto, Y, Wang, X, Takami, M, Yamada, A, Kamijo, R
  • Journal Title: J Dent Res 86(2) Pages: 163-168
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] S-Nitrosylation of human variant albumin Liprizzi(R410C) confers potent antibacterial and cytoprotective properties (2007)
  • Author(s): Ishima, Y, Sawa, T, Kragh-Hansen, U, Miyamoto, Y, Matsushita, S, Akaike, T, Otagairi, M
  • Journal Title: J Pharmacol Exp Ther 320(3) Pages: 969-977
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A new configuration device for mechanical stress culture (2007)
  • Author(s): Masuda, T, Takahashi, I, Arai, F, Anada, T, Miyamoto, Y, Fukuda, T, Suzuki, O
  • Journal Title: Proceedings of Internati.onal Symposium on Micro-Nano Mechatronics and Human Science Pages: 418-423
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction and characterization of the precursors committed to osteoclasts Induced by TNF-related activation-induced cytokine/receptor activator of NF-κB ligand (2006)
  • Author(s): Mochizuki, A, Takami, M, Kawawa, T, Suzumoto, R, Sasaki, T, Shiba, A, Tsukasaki, H, Zhao, B, Yasuhara, R, Suzawa, T, Miyamoto, Y, Choi, Y, Kamijo, R
  • Journal Title: J Immunol 177(7) Pages: 4360-4368
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Lysine-specific gingipain(Kgp) and lipopolysaccharide(LPS) synergistically induce osteoclast differentiation (2007)
  • Author(s): Yasuhara, R, Miyamoto, Y, Takami, M, Imamura, T, Kamijo, R
  • Organizer: 29th Annual Meeting of the American Society for Bone and Mineral Research
  • Place of Presentation: Honolulu, Hawaii, USA
  • Year and Date: 20070915-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Lysine-specific gingipain(Kgp) produce by Porphyromonas gingivalus and LPS synergistically induce osteoclast differentiation (2007)
  • Author(s): Yasuhara, R, Miyamoto, Y, Takami, M, Yoshimura, K, Kamijo, R
  • Organizer: 49th Annual Meeting of the Japanese Society Oral Biology
  • Place of Presentation: Sapporo
  • Year and Date: 20070830-31
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] n Reactive oxygen species-dependent change in pH is a key factor for matrix degradation in interleukin- 1-treated chondrocytes (2007)
  • Author(s): Yoshimura, K, Miyamoto, Y, Yasuhara, R, Kamijo, R
  • Organizer: 27th Regular Meeting of Showa University Dental Society
  • Place of Presentation: Tokyo
  • Year and Date: 2007-12-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Phagocyte-type NADPH-oxidase produces reactive oxygen species in chondrocytes in response to stimulation by interleukin-1β (2006)
  • Author(s): Yasuhara R
  • Organizer: 28th Annual Meeting of the American Society for Bone and Mineral Research
  • Place of Presentation: Philadelphia,PA,USA
  • Year and Date: 20060915-19
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Phagocyte-type NADPH-oxidase produces reactive oxygen species in mouse chondrocytes in response to stimulation by interleukin-ilβ (2006)
  • Author(s): Yasuhara, R, Miyamoto, Y, Yamada, A, Takami, M, Suzawa, T, Kamijo, R
  • Organizer: 28th Annual Meeting of the American Society for Bone and Mineral Research
  • Place of Presentation: Philadelphia, Pennsylvania, USA
  • Year and Date: 20060915-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Cell Adhesion signal regulates RANK Expression in osteoclast precursors (2006)
  • Author(s): Mochizuki, A, Takami, M, Kawawa, T, Miyamoto, Y, Suzawa, T, Yamada, A, Tomoyasu, S, Nakamaki, T, Kamijo, R
  • Organizer: 28th Annual Meeting of the American Society for Bone and Mineral Research
  • Place of Presentation: Philadelphia, Pennsylvania, USA
  • Year and Date: 20060915-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] The role of NADPH-oxidase derived reactive oxygen species in matrix degradation by chondrocytes (2006)
  • Author(s): Yasuhara, R, Miyamoto, Y, Yamada, A, Kamijo, R
  • Organizer: 20th IUBMB(International Union of Biochemistry and Molecular Biology) Congress
  • Place of Presentation: Kyoto
  • Year and Date: 20060618-23
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Nitric oxide suppresses proliferation and promotes mineralization of pulp cells (2006)
  • Author(s): Yasuhara, R, Suzawa, T, Miyamoto, Y, Wang, S, Tamami, M, Yamada, A, Matsumoto, K, Kamijo, R
  • Organizer: 28th Annual Meeting of the Japan Tissue Culture Society for Dental Research
  • Place of Presentation: Osaka
  • Year and Date: 2006-11-11
  • Description: 「研究成果報告書概要(欧文)」より
• [Remarks] 「研究成果報告書概要(和文)」より
  • URL: http://dents.showa-u.ac.jp/~dental/zdetail/detail.php?recordID=D02