4D ex-vivo imaging of NO signaling in osteocytes

• Project/Area Number: 24792281
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Orthodontic/Pediatric dentistry
• Research Institution: Okayama University
• Principal Investigator: ISHIHARA Yoshihito 岡山大学, 医歯(薬)学総合研究科, 助教 (70549881)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 歯科矯正学 / 骨細胞 / カルシウム / 細胞内カルシウム / NO / ギャップ結合

Research Abstract

Bone cells respond to mechanical stimuli by producing a variety of biological signals, and intracellular calcium (Ca2+) is an important secondary messenger controlling many cellular processes. Its variety of oscillatory signaling is thought to act in the regulation of many different cell functions including bone metabolisms. However, it has not been entirely clear the behavior and connectivity of the [Ca2+]i signaling networks in mechanotransduction between osteoblasts and osteocytes in integrated bone tissues due to complex bone matrices surrounding bone cells. To address this issue, we have developed a novel ex vivo live Ca2+ imaging system, which made it possible to observe the mechanical stress-induced Ca2+ oscillations.

Research Products

• [Journal Article] Oscillatory intracellular Ca^<2+> responses in living bone (2014)
  • Author(s): Ishihara Y , Sugawara Y, Kamioka H, Kawanabe N, Naruse K, Yamashiro T
  • Journal Title: J Oral Biosci Volume: (in press)
  • Peer Reviewed
• [Journal Article] Long-term stability of implant-anchored orthodontics in an adult patient with a Class II Division 2 malocclusion and a unilateral molar scissors-bite (2014)
  • Author(s): Ishihara Y , Kuroda S, Sugawara Y, Kurosaka H, Takano-Yamamoto T, Yamashiro T
  • Journal Title: Am J Orthod Dentofacial Orthop Volume: 145(4) Issue: 4 Pages: S100-S113
  • DOI: 10.1016/j.ajodo.2013.07.016
  • Peer Reviewed
• [Journal Article] 蛍光ライブイメージングを用いた骨組織中における細胞内カルシウムオシレーションの計測 (2014)
  • Author(s): 石原嘉人 , 菅原康代 , 上岡寛 , 川邉紀章 , 成瀬恵治 , 山城隆
  • Journal Title: 日本骨形態計測学会雑誌 Volume: 24巻 1号 Pages: 15-22
  • Peer Reviewed
• [Journal Article] Oscillatory intracellular Ca2+ responses in living bone. (2014)
  • Author(s): Yoshihito Ishihara, Yasuyo Sugawara, Hiroshi Kamioka, Noriaki Kawanabe, Keiji Naruse, Takashi Yamashiro
  • Journal Title: Journal of Oral Biosciences Volume: 56 Issue: 2 Pages: 49-53
  • DOI: 10.1016/j.job.2014.03.002
  • Peer Reviewed
• [Journal Article] 蛍光ライブイメージングを用いた骨組織中における細胞内カルシウムオシレーションの計測 (2014)
  • Author(s): 石原嘉人, 菅原康代, 上岡寛, 川邉紀章, 成瀬恵治, 山城隆
  • Journal Title: 日本骨形態計測学会雑誌 Volume: 24 Pages: 15-22
  • Peer Reviewed
• [Journal Article] Ex vivo real-time observation of Ca(2+) signaling in living bone in response to shear stress applied on the bone surface. (2013)
  • Author(s): Ishihara Y, Sugawara Y, Kamioka H, Kawanabe N, Hayano S, Balam TA, Naruse K, Yamashiro T.
  • Journal Title: Bone Volume: 53 Issue: 1 Pages: 204-215
  • DOI: 10.1016/j.bone.2012.12.002
  • Peer Reviewed
• [Journal Article] Indirect usage of miniscrew anchorage to intrude overerupted mandibular incisors in a Class II patient with a deep overbite (2013)
  • Author(s): Ishihara Y , Kuroda S, Sugawara Y, Balam TA, Takano-Yamamoto T, Yamashiro T
  • Journal Title: Am J Orthod Dentofac Orthop Volume: 143(4) Issue: 4 Pages: S113-S124
  • DOI: 10.1016/j.ajodo.2012.09.001
  • Peer Reviewed
• [Journal Article] Extraction of the lateral incisors to treat maxillary protrusion : Quantitative evaluation of the stomatognathic functions (2013)
  • Author(s): Ishihara Y , Kuroda S, Sumiyoshi K, Takano-Yamamoto T, Yamashiro T
  • Journal Title: Angle Orthod Volume: 83(2) Issue: 2 Pages: 341-354
  • DOI: 10.2319/042412-343.1
  • Peer Reviewed
• [Journal Article] The early mouse 3D osteocyte network in the presence and absence of mechanical loading. (2013)
  • Author(s): Sugawara Y, Kamioka H, Ishihara Y, Fujisawa N, Kawanabe N, Yamashiro T.
  • Journal Title: Bone Volume: 52 Issue: 1 Pages: 189-196
  • DOI: 10.1016/j.bone.2012.09.033
  • Peer Reviewed
• [Journal Article] Gap junction-mediated communication in human periodontal ligament cells (2013)
  • Author(s): Kato R, Ishihara Y, Kawanabe N, Sumiyoshi K, Yoshikawa Y, Nakamura M, Imai Y, Yanagita T, Fukushima H, Kamioka H, Takano-Yamamoto T and Yamashiro T
  • Journal Title: JDR Volume: 92(7) Issue: 7 Pages: 635-40
  • DOI: 10.1177/0022034513489992
  • Peer Reviewed
• [Journal Article] In situ imaging of the autonomous intracellular Ca2+ oscillations of osteoblasts and osteocytes in bone (2012)
  • Author(s): Ishihara, Y., Sugawara Y., et al.
  • Journal Title: Bone Volume: 50 Issue: 4 Pages: 842-852
  • DOI: 10.1016/j.bone.2012.01.021
  • Peer Reviewed
• [Journal Article] Roles of Heparan Sulfate Sulfation in Dentinogenesis (2012)
  • Author(s): Hayano S, Kurosaka H, Yanagita T, Kalus I, Milz F, Ishihara Y, et al
  • Journal Title: J Biol Chem Volume: (印刷中) Issue: 15 Pages: 12217-12229
  • DOI: 10.1074/jbc.m111.332924
  • Peer Reviewed
• [Journal Article] Stage-specific embryonic antigen-4 identifies human dental pulp stemcells (2012)
  • Author(s): Kawanabe N, Murata S, Fukushima H, Ishihara Y, et al
  • Journal Title: Exp Cell Res Volume: 318 Issue: 5 Pages: 453-483
  • DOI: 10.1016/j.yexcr.2012.01.008
  • Peer Reviewed
• [Journal Article] SSEA-4 is a marker of human deciduous periodontal ligament stem cells. (2012)
  • Author(s): Fukushima H., Kawanabe N., Murata S., et al.
  • Journal Title: Journal of Dental Research Volume: 91 Issue: 10 Pages: 955-960
  • DOI: 10.1177/0022034512458123
  • Peer Reviewed
• [Journal Article] Promotion of Ccn2 expression and osteoblastic differentiation by actin polymerization, which is induced by laminar fluid flow stress (2012)
  • Author(s): Honjo T, Kubota S, Kamioka H, Sugawara Y, Ishihara Y, Yamashiro T, Takigawa M, Takano-Yamamoto T
  • Journal Title: J Cell Commun Signal Volume: 6(4) Issue: 4 Pages: 225-32
  • DOI: 10.1007/s12079-012-0177-z
  • Peer Reviewed
• [Journal Article] Functional improvements following orthodontic-surgical reconstruction in a patient affected by multiple maxillofacial fractures : a case report (2012)
  • Author(s): Ishihara Y , Kuroda S, Nishiyama A, Sasaki A, Takano-Yamamoto T, Yamashiro T
  • Journal Title: Am J Orthod Dentofac Orthop Volume: 142(4) Issue: 4 Pages: 534-535
  • DOI: 10.1016/j.ajodo.2011.02.029
  • Peer Reviewed
• [Presentation] 3 次元蛍光ライブイメージングで見る骨組織内の細胞応答 (2013)
  • Author(s): 石原嘉人
  • Organizer: 第8回東北大学大学院歯学研究科顎口腔矯正学分野セミナー
  • Place of Presentation: 仙台
  • Year and Date: 2013-05-08
• [Presentation] 生きた骨組織中で起こる細胞応答のイメージング (2013)
  • Author(s): 石原嘉人 , 菅原康代 , 上岡寛 , 成瀬恵治 , 山城隆
  • Organizer: 第33回骨形態計測学会
  • Place of Presentation: 浜松
• [Presentation] 骨組織中でのライブイメージングを用いた自律性細胞内カルシウムオシレーションの検討 (2013)
  • Author(s): 石原嘉人
  • Organizer: 第55回歯科基礎医学会学術大会・総会
  • Place of Presentation: 岡山
• [Presentation] Gap junction-mediated communication in human periodontal ligament cells (2013)
  • Author(s): Kato R, Ishihara Y , Kawanabe N, Sumiyoshi K, Yoshikawa Y, Nakamura M, Imai Y, Yanagita T, Fukushima H, Kamioka H, Takano-Yamamoto T, Yamashiro T
  • Organizer: NIH-Tohoku Univerity- JSPS Symposium Sendai
  • Place of Presentation: Japan
• [Presentation] Ex vivo real-time observation of Ca^<2+> signaling in living bone in response to shear stress applied on the bone surface (2013)
  • Author(s): Ishihara Y , Sugawara Y, Kamioka H, Kawanabe N, Hayano S, Balam TA, Naruse K, Yamashiro T
  • Organizer: 2nd Joint Meeting of the International Bone and Mineral Society and the Japanese Society for Bone and Mineral Research Kobe
  • Place of Presentation: Japan
• [Presentation] The early mouse 3D osteocyte network in the presence and absence of mechanical loading (2013)
  • Author(s): Sugawara Y, Kamioka H, Ishihara Y , Fujisawa N, Kawanabe N, Yamashiro T
  • Organizer: 2nd Joint Meeting of the International Bone and Mineral Society and the Japanese Society for Bone and Mineral Research Kobe
  • Place of Presentation: Japan
• [Presentation] Ex-vivo imaging of fluid flow-induced intracellular Ca^<2+> oscillations of osteoblasts and osteocytes in bone (2013)
  • Author(s): Ishihara Y , Sugawara Y, Kamioka H, Kawanabe N , Yamashiro T
  • Organizer: 89th Congress of the European orthodontic society Reykjavik
  • Place of Presentation: Iceland
• [Presentation] ヒト歯根膜細胞間ギャップ結合の機能性および調節機構は低酸素環境下において抑制的に調節される (2013)
  • Author(s): 加藤龍史 , 石原嘉人 , 川邉紀章 , 上岡寛 , 山本照子 , 山城隆
  • Organizer: 第31回日本骨代謝学会学術集会
  • Place of Presentation: 神戸
• [Presentation] ヒト歯根膜細胞におけるギャップ結合を介した細胞間コミュニケーションの検討 (2013)
  • Author(s): 加藤龍史 , 石原嘉人 , 上岡寛 , 山本照子 , 山城隆
  • Organizer: 第55回歯科基礎医学会学術大会・総会
  • Place of Presentation: 岡山
• [Presentation] Ex vivo real-time observation of Ca2+ signaling in living bone in response to shear stress applied on the bone surface. (2013)
  • Author(s): Yoshihito Ishihara, Yasuyo Sugawara, Hiroshi Kamioka, Noriaki Kawanabe, Satoru Hayano, Tarek A. Balam, Keiji Naruse, Takashi Yamashiro.
  • Organizer: 2nd Joint Meeting of the International Bone and Mineral Society and the Japanese Society for Bone and Mineral Research
  • Place of Presentation: Kobe, Japan
• [Presentation] Ex-vivo imaging of fluid flow-induced intracellular Ca2+ oscillations of osteoblasts and osteocytes in bone. (2013)
  • Author(s): Yoshihito Ishihara, Yasuyo Sugawara, Hiroshi Kamioka, Noriaki Kawanabe, Takashi Yamashiro.
  • Organizer: 89th Congress of the European orthodontic society
  • Place of Presentation: Reykjavik, Iceland
• [Presentation] The early mouse 3D osteocyte network in the presence and absence of mechanical loading. (2013)
  • Author(s): Sugawara Y, Kamioka H, Ishihara Y, Fujisawa N, Kawanabe N, Yamashiro T.
  • Organizer: 2nd Joint Meeting of the International Bone and Mineral Society and the Japanese Society for Bone and Mineral Research
  • Place of Presentation: Kobe, Japan
• [Presentation] In situ imaging of the autonomous intracellular Ca^<2+> oscillations of osteoblasts and osteocytes in bone (2012)
  • Author(s): Ishihara Y , Sugawara Y, Kamioka H, Kawanabe N, Naruse K, Yamashiro T
  • Organizer: 1st Asia-Pacific Bone and Mineral Research Meeting with ANZBMS 22nd Annual Scientific Meeting
  • Place of Presentation: Perth, Australia
• [Presentation] Stage-specific embryonic antigen-4 is a marker of human deciduous periodontal ligament stem cells (2012)
  • Author(s): Kawanabe N, Fukushima H, Murata S, Ishihara Y , Yanagita T, Balam TA, Yamashiro T
  • Organizer: The ASBMR 34th Annual Meeting Minneapolis
  • Place of Presentation: USA
• [Presentation] Live imaging of fluid flow-induced Ca^<2+> signaling of osteoblasts and osteocytes in bone : implications for gap junctional intercellular communication (2012)
  • Author(s): Ishihara Y , Sugawara Y, Kamioka H, Kawanabe N, Naruse K, Yamashiro T
  • Organizer: The ASBMR 34th Annual Meeting Minneapolis
  • Place of Presentation: USA
• [Presentation] 蛍光ライブイメージングを用いた骨組織中における自律性細胞内カルシウムオシレーションの計測 (2012)
  • Author(s): 石原嘉人 , 菅原康代 , 上岡寛 , 川邉紀章 , 成瀬恵治 , 山城隆
  • Organizer: 第32回骨形態計測学会
  • Place of Presentation: 大阪
• [Presentation] 生体ライブイメージングを用いた骨組織中における自律性細胞内カルシウムオシレーションの解析 (2012)
  • Author(s): 石原嘉人 , 菅原康代 , 上岡寛 , 山城隆
  • Organizer: 第30回日本骨代謝学会学術集会
  • Place of Presentation: 東京
• [Presentation] 低酸素環境下におけるヒト歯根膜細胞間ギャップ結合の機能性および調節機構の検討 (2012)
  • Author(s): 加藤龍史 , 石原嘉人 , 上岡寛 , 山本照子 , 山城隆
  • Organizer: 第54回歯科基礎医学会学術大会・総会
  • Place of Presentation: 福島
• [Presentation] イノシトール三リン酸は生きた骨組織中における細胞内カルシウム応答の必須因子である (2012)
  • Author(s): 石原嘉人 , 菅原康代 , 上岡寛 , 川邉紀章 , 柳田剛士 , 住吉久美 , 山城隆
  • Organizer: 第71回日本矯正歯科学会大会
  • Place of Presentation: 盛岡
• [Presentation] 電子トモグラフィー法による骨芽細胞のコラーゲン排出過程の観察 (2012)
  • Author(s): 保崎留美子 , 上岡寛 , 今井裕一 , 石原嘉人 , 菅原康代 , 山城隆
  • Organizer: 第71回日本矯正歯科学会大会
  • Place of Presentation: 盛岡
• [Presentation] Stage-specific embryonic antigen-4 is a marker of human deciduous periodontal ligament stem cells (2012)
  • Author(s): Noriaki Kawanabe, Hiroaki Fukushima, Satoko Murata, Yoshihito Ishihara, Takeshi Yanagita, Tarek A. Balam, Takashi Yamashiro
  • Organizer: The ASBMR 34th Annual Meeting
  • Place of Presentation: Minneapolis, USA
• [Presentation] Live imaging of fluid flow-induced Ca2+ signaling of osteoblasts and osteocytes in bone: implications for gap junctional intercellular communication (2012)
  • Author(s): Yoshihito Ishihara, Yasuyo Sugawara, Hiroshi Kamioka, Noriaki Kawanabe, Keiji Naruse, Takashi Yamashiro
  • Organizer: The ASBMR 34th Annual Meeting
  • Place of Presentation: Minneapolis, USA
• [Presentation] ヒト歯根膜細胞におけるギャップ結合を介した細胞間伝達は低酸素環境下において抑制的に調節される (2012)
  • Author(s): 加藤 龍史, 石原 嘉人, 川邉 紀章, 住吉 久美, 吉川 祐介, 今井 裕一, 上岡 寛, 山本 照子, 山城 隆
  • Organizer: 第71回日本矯正歯科学会大会
  • Place of Presentation: 盛岡
• [Presentation] 低酸素環境下におけるヒト歯根膜細胞間ギャップ結合の機能性および調節機構の検討 (2012)
  • Author(s): 加藤 龍史, 石原 嘉人, 上岡 寛, 山本 照子, 山城 隆
  • Organizer: 第54回歯科基礎医学会学術大会・総会
  • Place of Presentation: 郡山
• [Presentation] In situ imaging of the autonomous intracellular Ca2+ oscillations of osteoblasts and osteocytes in bone (2012)
  • Author(s): Yoshihito Ishihara, Yasuyo Sugawara, Hiroshi Kamioka, Noriaki Kawanabe, Keiji Naruse, Takashi Yamashiro
  • Organizer: 1st Asia-Pacific Bone and Mineral Research Meeting with ANZBMS 22nd Annual Scientific Meeting
  • Place of Presentation: Perth, Australia
• [Presentation] 3次元蛍光ライブイメージングで見る骨組織内の細胞応答
  • Author(s): 石原嘉人
  • Organizer: 第8回東北大学大学院歯学研究科顎口腔矯正学分野セミナー
  • Place of Presentation: 仙台
  • Invited
• [Book] The BONE 特集『骨バイオイメージング』 骨細胞の機能イメージング (2014)
  • Author(s): 石原嘉人, 上岡寛
  • Publisher: メディカルレビュー社(in press)
• [Book] The BONE特集『骨バイオイメージング』 骨細胞の機能イメージング (2014)
  • Author(s): 石原嘉人, 上岡寛
  • Total Pages: 6
  • Publisher: メディカルレビュー社