Elucidation of the calcified promotion mechanism of osteoblasts by the carbon nanotubes

• Project/Area Number: 25462365
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Shinshu University
• Principal Investigator: HANIU Hisao 信州大学, 学術研究院医学系, 准教授 (30252050)
• Co-Investigator(Kenkyū-buntansha): USUI Yuki 信州大学, 医学部附属病院, 特任研究員 (00467169) ; TSUKAHARA Tamotsu 長崎大学, 医歯(薬)学総合研究科, 准教授 (00529943)
• Co-Investigator(Renkei-kenkyūsha): SAITO Naoto 信州大学, 学術研究院保健学系, 教授 (80283258)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 骨・軟骨代謝学 / バイオマテリアル / カーボンナノチューブ / 骨芽細胞

Outline of Final Research Achievements

We examined the relationship between Multi-walled carbon nanotubes (MWCNTs) and a MC3T3-E1 preosteoblast cell line. MWCNTs were combined with three different dispersants (FBS, gelatin, or CMC). Cell viability was measured under various culture conditions and morphological analysis was performed. We also assessed several osteoblast differentiation markers. Inhibition of cell proliferation was witnessed only under specific conditions, such as when MC3T3-E1 not cultured in calcification medium was exposed to MWCNTs dispersed in gelatin. Furthermore, cell proliferation increased with all dispersants when the cells were cultured in calcification medium. In morphological observations, whereas the cells displaying proliferation inhibition had internalized MWCNTs into the cytoplasm, the biomaterial was found to be adhered to the cell membrane in other conditions. MWCNTs up-regulated the expression of one marker of osteoblast differentiation, regardless of the dispersant.

Research Products

• [Journal Article] The role of autophagy as a mechanism of toxicity induced by multi-walled carbon nanotubes in human lung cells. (2015)
  • Author(s): Tsukahara T, Matsuda Y, Haniu H.
  • Journal Title: Int J Mol Sci. Volume: 16 Issue: 1 Pages: 40-48
  • DOI: 10.3390/ijms16010040
  • NAID: 120007110640
  • Peer Reviewed / Open Access
• [Journal Article] Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response (2013)
  • Author(s): Haniu H, Saito N, Matsuda Y, Tsukahara T, Maruyama K, Usui Y, Aoki K, Takanashi S, Kobayashi S, Nomura H, Okamoto M, Shimizu M, Kato H
  • Journal Title: Toxicol in Vitro Volume: 27 Issue: 6 Pages: 1679-1685
  • DOI: 10.1016/j.tiv.2013.04.012
  • NAID: 120007110519
  • Peer Reviewed
• [Journal Article] Highly purified, multi-wall carbon nanotubes induce light-chain 3B expression in human lung cells. (2013)
  • Author(s): Tsukahara T, Matsuda Y, Usui Y, Haniu H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 440 Issue: 2 Pages: 348-53
  • DOI: 10.1016/j.bbrc.2013.09.089
  • NAID: 120007100190
  • Peer Reviewed
• [Presentation] Effect of multi-walled carbon nanotubes in three different dispersants on MC3T3-E1 cell line proliferation. (2015)
  • Author(s): Haniu H.
  • Organizer: 5th International Conference on Nanotek & Expo.
  • Place of Presentation: San Antonio, USA
  • Year and Date: 2015-11-16
  • Int'l Joint Research
• [Presentation] A capacity of porous multi-walled carbon nanotube blocks as scaffold of bone regeneration. (2015)
  • Author(s): 40.Tanaka M, Saito N, Zhang M, Sato Y, Haniu H, Takizawa T, Nomura H, Kobayashi S, Okamoto M, Takanashi S, Aoki K, Usui Y, Kato H.
  • Organizer: 12th Bone Biology Forum
  • Place of Presentation: San Antonio, USA
  • Year and Date: 2015-08-03
  • Int'l Joint Research
• [Presentation] The effect of carbon nanotube blocks on bone regeneration. (2015)
  • Author(s): 39.Tanaka M, Saito N, Zhang M, Sato Y, Haniu H, Takizawa T, Nomura H, Kobayashi S, Okamoto M, Takanashi S, Aoki K, Usui Y, Kato H.
  • Organizer: Sixth Symposium on Carbon Nanomaterials Biology, Medicine & Toxicology
  • Place of Presentation: Nagoya, Japan
  • Year and Date: 2015-06-28
  • Int'l Joint Research
• [Presentation] Evaluation of Multi-walled Carbon Nanotubes Using the MC3T3-E1 Cell Line. (2015)
  • Author(s): Haniu H, Saito N, Maruyama K, Matsuda N, Tanaka M, Aoki K, Okamoto M, Kobayashi S, Nomura H, Takizawa T, Ohishi A, Usui Y, Kato H.
  • Organizer: TechConnect World Innovation Conference & Expo-2015
  • Place of Presentation: Washinton, DC, USA
  • Year and Date: 2015-06-14
  • Int'l Joint Research
• [Presentation] Endocytosis of MWCNT in BEAS-2B cells are affected by the culture medium type. (2013)
  • Author(s): Hisao Haniu, Naoto Saito, Yoshikazu Matsuda, Kayo Maruyama, Tamotsu Tsukahara, Yuki Usui, Kaoru Aoki, Seiji Takanashi, Shinsuke Kobayashi, Hiroki Nomura, Masanori Okamoto, Masayuki Shimizu, Hiroyuki Kato.
  • Organizer: 6th International Symposium on Nanotechnology, Occupational and Environmental Health
  • Place of Presentation: 愛知県名古屋市
• [Presentation] An innovative method to evaluate biodistribution and kinetics of carbon nanotubes using CNT peapods (2013)
  • Author(s): Shinsuke Kobayashi, Shuji Tsuruoka, Yuki Usui, Hisao Haniu, Kaoru Aoki, Norio Ishigaki, Masayuki Shimizu, Seiji Takanashi, Masanori Okamoto, Hiroki Nomura, Hiroyuki Kato, Naoto Saito
  • Organizer: 6th International Symposium on Nanotechnology, Occupational and Environmental Health
  • Place of Presentation: 愛知県名古屋市
• [Presentation] Intraarticular reaction and secretion of inflammatory chemokines to multi wall carbon nanotubes
  • Author(s): Hiroki Nomura, Hisao Haniu, Seiji Takanashi, Shinsuke Kobayashi, Kaoru Aoki, Kayo Maruyama, Yuki Usui, Naoto Saito
  • Organizer: 6th International Symposium on Nanotechnology, Occupational and Environmental Health
  • Place of Presentation: 愛知県名古屋市
• [Presentation] Drug delivery system by carbon nanotubes for sarcoma cells
  • Author(s): Kaoru Aoki, Hisao Haniu, Masayuki Shimizu, Seiji Takanashi, Masanori Okamoto, Shinsuke Kobayashi, Hiroki Nomura, Yuki Usui, Naoto Saito
  • Organizer: 6th International Symposium on Nanotechnology, Occupational and Environmental Health
  • Place of Presentation: 愛知県名古屋市
• [Presentation] 培養液による多層カーボンナノチューブのBEAS-2B細胞におけるバイオレ スポンスへの影響
  • Author(s): 羽二生久夫、齋藤直人、松田佳和、丸山佳与、薄井雄企、青木薫、高梨誠司、 小林伸輔、野村博紀、岡本正則、清水政幸、加藤博之
  • Organizer: 第40回日本毒性学会学術年会
  • Place of Presentation: 千葉県千葉市