A trial approach for tooth regeneration by three-layer sandwich culture using neural crest-derived cells

• Project/Area Number: 24592815
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Showa University
• Principal Investigator: SUZAWA Tetsuo 昭和大学, 歯学部, 講師 (60271285)
• Co-Investigator(Kenkyū-buntansha): KAMIJO Ryutaro 昭和大学, 歯学部, 教授 (70233939)
• Co-Investigator(Renkei-kenkyūsha): OSUMI Noriko 東北大学, 医学系研究科, 教授 (00220343) ; KUNISADA Takahiro 岐阜大学, 医学系研究科, 教授 (30205108)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 再生医学 / 神経堤由来細胞 / 組織幹細胞 / 発生・分化 / 歯胚再生 / 神経堤細胞

Outline of Final Research Achievements

Neural crest cells in the embryo migrate to reach target sites as neural crest-derived cells (NCDCs) where they differentiate into a variety of derivatives. With the aim of facilitating tooth regeneration by using non-dental-derived cells, we analyzed the characteristics of neural crest-derived cells (NCDCs) in adult tissues. We used P0-Cre/CAG-CAT-EGFP transgenic mice (P0 mice), in which NCDCs express green fluorescent protein (GFP). GFP-positive cells separated using flow cytometry expressed a high level of endothelin receptor type B (Ednrb) by DNA microarray. These GFP-positive cells were capable of proliferating into spheres and exist in the oral mucosa. They also were increased by endothelin 3, a ligand of Ednrb. Furthermore, GFP-positive cells had the capability to differentiate into various cell lineages including osteoblast-like cells and adipocyte-like cells. Therefore, adult oral NCDCs may be a useful cell source for regenerative medicine, including tooth regeneration.

Research Products

• [Journal Article] Bone Response to Static Compressive Stress at Bone-Implant Interface: The Pilot Study of Critical Static Compressive stress. (2015)
  • Author(s): Ikumi N, Suzawa T, Yoshimura K, Kamijo R.
  • Journal Title: Int J Oral Maxillofac Implants Volume: - Issue: 4 Pages: 827-833
  • DOI: 10.11607/jomi.3715
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Microscopic study on resorption of β-tricalcium phosphate materials by osteoclasts. (2015)
  • Author(s): Matsunaga A, Takami M, Irié T, Mishima K, Inagaki K, Kamijo R.
  • Journal Title: Cytotechnology Volume: -
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] In-situ quasi-static and dynamic nanoindentation tests on calcified nodules formed by osteoblasts: Implication of glucocorticoids responsible for osteoblast calcification (2015)
  • Author(s): Miyamoto S., Miyamoto Y., Shibata Y., Yoshimura K., Izumida E., Suzuki H., Miyazaki T., Maki K., Kamijo R.
  • Journal Title: Acta Biomaterialia Volume: １２ Pages: 216-226
  • DOI: 10.1016/j.actbio.2014.10.038
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cdc42 is critical for cartilage development during endochondral ossification. (2015)
  • Author(s): Suzuki W, Yamada A, Aizawa R, Suzuki D, Kassai H, Harada T, Nakayama M, Nagahama R, Maki K, Takeda S, Yamamoto M, Aiba A, Baba K, Kamijo R
  • Journal Title: Endocrinology Volume: 156 Issue: 1 Pages: 314-322
  • DOI: 10.1210/en.2014-1032
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Porphyromonas gingivalis-derived lysine gingipain enhances osteoclast differentiation induced by tumor necrosis factor and interleukin-1 but suppresses that by interleukin-17: Importance of proteolytic degradation of osteoprotegerin by lysine gingipain. (2014)
  • Author(s): Tomohito Akiyama, Yoichi Miyamoto, Kentaro Yoshimura, Atsushi Yamada, Masamichi Takami, Tetsuo Suzawa, Marie Hoshino, Takahisa Imamura, Chie Akiyama, Rika Yasuhara, Kenji Mishima, Toshifumi Maruyama, Chikara Kohda, Kazuo Tanaka, Jan Potempa, Hisataka Yasuda, Kazuyoshi Baba, Ryutaro Kamijo.
  • Journal Title: J.Biol.Chem. Volume: 289 Issue: 22 Pages: 1-35
  • DOI: 10.1074/jbc.m113.520510
  • Peer Reviewed / Open Access
• [Journal Article] Effects of antibody to receptor activator of nuclear factor κ-B ligand on inflammation and cartilage degradation in collagen antibody-induced arthritis in mice. (2014)
  • Author(s): Funato S, Matsunaga A, Oh K, Miyamoto Y, Yoshimura K, Tanaka J, Suzuki D, Uyama R, Suzuki H, Mishima K, Nakamura M, Namiki O, Baba K, Inagaki K, Kamijo R.
  • Journal Title: J Negat Results Biomed. Volume: 13 Issue: 1 Pages: 18-18
  • DOI: 10.1186/s12952-014-0018-0
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of gene expression profile of neural crest-derived cells isolated from submandibular glands of adult mice. (2014)
  • Author(s): Takahashi M, Suzawa T, Yamada A, Yamaguchi T, Mishima K, Osumi N, Maki K, Kamijo R
  • Journal Title: BBRC Volume: 30 Issue: 2 Pages: 1925-1937
  • DOI: 10.1016/j.bbrc.2014.02.130
  • Peer Reviewed
• [Journal Article] Nanomechanical properties and molecular structures of in vitro mineralized tissues on anodically oxidized titanium surfaces. (2014)
  • Author(s): Jang, I-. K., Tanaka, R., Wurihan, Suzuki, D., Shibata Y., Fujisawa, N., Tanimoto, Y., Ogura, K., Kamijo, R., Miyazaki, T.
  • Journal Title: Nanomedicine Volume: 10 Issue: 3 Pages: 629-637
  • DOI: 10.1016/j.nano.2013.09.007
  • Peer Reviewed
• [Journal Article] Synthetic Octacalcium Phosphate Enhanced Reparative Dentin Formation via Induction of Odontoblast Differentiation. (2013)
  • Author(s): X Wang, T Suzawa, T Miyauchi, B Zhao, R Yasuhara, T Anada, N Nakamura, O Suzuki, R Kamijo.
  • Journal Title: J Tissue Eng Regen Med, Volume: in press Issue: 11 Pages: 1310-1320
  • DOI: 10.1002/term.1669
  • Peer Reviewed
• [Journal Article] Downregulation of carbonic anhydrase IX promotes Col10a1 expression in chondrocytes (2013)
  • Author(s): Maruyama T, Miyamoto Y, Yamamoto G, Yamada A, Yoshimura K, Suzawa T, Takami M, Akiyama T, Hoshino M, Iwasa F, Ikumi N, Tachikawa T, Mishima K, Baba K, Kamijo R
  • Journal Title: PLoS ONE Volume: Vol.8 Issue: 2 Pages: e56984-e56984
  • DOI: 10.1371/journal.pone.0056984
  • Peer Reviewed
• [Journal Article] Octacalcium phosphate suppresses chondrogenic differentiation of ATDC5 cells. (2013)
  • Author(s): I Shibuya, K Yoshimura, Y Miyamoto, A Yamada, M Takami, T Suzawa, D Suzuki, N Ikumi, F Hiura, T Anada, O Suzuki, R Kamijo.
  • Journal Title: Cell Tissue Res Volume: 352 Issue: 2 Pages: 401-412
  • DOI: 10.1007/s00441-012-1548-8
  • Peer Reviewed
• [Journal Article] BMP2 differentially regulates the expression of Gremlin1 and Gremlin2, the negative regulators of BMP function, during osteoblast differentiation (2012)
  • Author(s): Suzuki D, Yamada A, Aizawa R, Funato S, Matsumoto T, Suzuki W, Takami M, Miyamoto Y, Suzawa T, Yamamoto M, Baba K, Kamijo R
  • Journal Title: Calcif Tissue Int Volume: 91 Issue: 1 Pages: 88-96
  • DOI: 10.1007/s00223-012-9614-5
  • Peer Reviewed
• [Presentation] 8-Nitro-guanosine 3’,5’-cyclic monophosphate mediates elongation of the growth plate cartilage in mice. (2014)
  • Author(s): Hoshino M, Miyamoto Y, Kameko K, Yoshimura K, Suzuki D, Akaike T, Baba K, Kamijo R
  • Organizer: The 2014 ASCB/IFCB Meeting
  • Place of Presentation: Philadelphia, PE, U.S.A.
  • Year and Date: 2014-12-06 – 2014-12-11
• [Presentation] Endogenously produced persulfides enhance bone elongation. (2014)
  • Author(s): Hoshino M, Kaneko K, Miyamoto Y, Yoshimura K, Suzuki D, Akaike T, Chikazu D, Baba K, Kamijo R
  • Organizer: Janapnese Association for Dental Research 62th Annual Meeting
  • Place of Presentation: 大阪
  • Year and Date: 2014-12-04 – 2014-12-05
• [Presentation] 成体マウス顎下腺から分離した神経堤由来細胞の解析 (2014)
  • Author(s): 高橋正皓、須澤徹夫、山田篤、山口徹太郎、槇宏太郎、上條竜太郎
  • Organizer: 第51回日本口腔組織培養学会学術大会
  • Place of Presentation: 福岡
  • Year and Date: 2014-11-15
• [Presentation] ヒトDown症乳歯歯根膜由来細胞のSDF-1発現解析 (2014)
  • Author(s): 浅川剛吉，宮本洋一，吉村健太郎，長谷川智一，山下一恵，嘉手納未季，馬目瑤子，栗谷未来，上條竜太郎，船津手敬弘
  • Organizer: 第31回 日本障害者歯科学会総会および学術大会
  • Place of Presentation: 仙台
  • Year and Date: 2014-11-14 – 2014-11-16
• [Presentation] 永久歯先天欠如の新規原因遺伝子 (2014)
  • Author(s): 山口徹太郎，細道一善，矢野圭介，芳賀秀郷，方山光朱，中脇貴俊，富田大介，中村雅典，上條竜太郎，井ノ上逸朗，槇 宏太郎
  • Organizer: 第73回日本矯正歯科学会大会
  • Place of Presentation: 千葉
  • Year and Date: 2014-10-20 – 2014-10-22
• [Presentation] Cdc42は口蓋形成過程において必須の遺伝子である (2014)
  • Author(s): 中山睦子，山田 篤，佐藤友紀，上條竜太郎，槇 宏太郎
  • Organizer: 第73回日本矯正歯科学会大会
  • Place of Presentation: 千葉
  • Year and Date: 2014-10-20 – 2014-10-22
• [Presentation] 破骨細胞の前駆細胞形成における骨芽細胞と炎症性刺激の役割 (2014)
  • Author(s): 高見正道，榎本拓哉，山本松男，上條竜太郎
  • Organizer: 第56回歯科基礎医学会学術大会・総会 サテライトシンポジウム
  • Place of Presentation: 福岡
  • Year and Date: 2014-09-25
• [Presentation] 成体マウス顎下腺における神経堤由来細胞の解析 (2014)
  • Author(s): 高橋正皓、須澤徹夫、山田篤、上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] 骨髄、脾臓、血液中に存在する破骨細胞分化能をもつ細胞の解析 (2014)
  • Author(s): 榎本拓哉，高見正道，山本松男，上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] 培養破骨細胞によるb-TCP製人工骨補填剤オスフェリオンの吸収 (2014)
  • Author(s): 松永朗裕，高見正道，上條竜太郎，稲垣克記
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] Cdc42は軟骨形成とそれに続く軟骨内骨化に必須である (2014)
  • Author(s): 鈴木 航，山田 篤，相澤 怜，鈴木 大，竹田 秀，山本松男，上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] ケトン体は骨芽細胞機能を調節する (2014)
  • Author(s): 齊藤彰大，吉村健太郎，宮本洋一，山本松男，上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] グルココルチコイド系ステロイドは骨芽細胞によるアパタイト含有石灰化物形成とそのかたさを向上させる (2014)
  • Author(s): 宮本 尚，宮本洋一，吉村健太郎，槇 宏太郎，上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] 骨巨細胞腫細胞に対するDenosumabの効果 (2014)
  • Author(s): 渋谷 勲，高見正道，松本光史，榎本拓哉，松永朗裕，浦野絵里，上條竜太郎
  • Organizer: 第32回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2014-07-24 – 2014-07-26
• [Presentation] 培養破骨細胞によるβ-TCP製人工骨補填材オスフェリンの吸収 (2014)
  • Author(s): 松永朗裕，高見正道，上條竜太郎，稲垣克記
  • Organizer: 第14回東京骨関節フォーラム
  • Place of Presentation: 東京
  • Year and Date: 2014-07-19
• [Presentation] 歯周病原菌の毒素による破骨細胞分化の促進 (2014)
  • Author(s): 秋山智人，安原理佳，宮本洋一，今村隆寿，馬場一美，美島健二，上條竜太郎
  • Organizer: 第14回東京骨関節フォーラム
  • Place of Presentation: 東京
  • Year and Date: 2014-07-19
• [Presentation] Modulation of osteoblast by ketone bodies. (2014)
  • Author(s): Saito A, Yoshimura K, Miyamoto Y, Yamamoto M, Kamijo R
  • Organizer: 92nd General Session & Exhibition of IADR
  • Place of Presentation: Cape Town, South Africa
  • Year and Date: 2014-06-25 – 2014-06-28
• [Presentation] アルジネート印象材から創製したケイ酸セメントのラット培養歯髄細胞に対する影響について (2014)
  • Author(s): 増田宜子，山口暢章，山田嘉重，宮本洋一，上條竜太郎，宮﨑 隆
  • Organizer: 日本歯科保存学会2014年度春期学術大会（第140回）
  • Place of Presentation: 大津
  • Year and Date: 2014-06-19 – 2014-06-20
• [Presentation] 神経堤由来の毛包細胞を用いた骨芽細胞分化誘導． (2013)
  • Author(s): 森澤絵里，高見正道，須澤徹夫，馬場一美，大隅典子，上條竜太郎
  • Organizer: 第68回NPO法人日本口腔科学会学術集会
  • Place of Presentation: 宇都宮
• [Presentation] 成体組織に存在する神経堤由来細胞の効率的な分離法を目指した遺伝子発現パターンの解析． (2013)
  • Author(s): 高橋正晧，須澤徹夫，山田篤、山口徹太郎、上條竜太郎、槇宏太郎
  • Organizer: 第72回日本矯正歯科学会大会
  • Place of Presentation: 松本
• [Presentation] 毛包に存在する神経堤由来細胞を用いた骨芽細胞分化誘導． (2013)
  • Author(s): 森澤絵里，高見正道，須澤徹夫，馬場一美，大隅典子，上條竜太郎
  • Organizer: 日本口腔組織培養学会設立50周年記念学術大会・総会
  • Place of Presentation: 東京
• [Presentation] 成体組織における神経堤由来細胞の遺伝子発現解析． (2013)
  • Author(s): 高橋正晧，須澤徹夫，山田篤、山口徹太郎、上條竜太郎、槇宏太郎
  • Organizer: 第33回昭和歯学会例会
  • Place of Presentation: 東京
• [Presentation] Characteristic gene expression profile of neural crest-derived cells in adult mouse tissues. (2013)
  • Author(s): Takahashi M, Suzawa T, Yamada A, Maki K, Kamijo R.
  • Organizer: American Society for Cell Biology 2013 Annual Meeting
  • Place of Presentation: New Orleans, USA
• [Presentation] 毛包内の神経堤由来細胞による象牙芽細胞分化誘導 (2012)
  • Author(s): 森澤絵里，須澤徹夫，宮内知彦，高見正道，山田篤，宮本洋一，林竜平，西田幸二，大隅典子，馬場一美，上條竜太郎
  • Organizer: 第11回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜（横浜）
• [Presentation] 毛包から採取した神経堤由来細胞による象牙芽細胞分化誘導 (2012)
  • Author(s): 森澤絵里，須澤徹夫，宮内知彦，高橋正皓，槇宏太郎，馬場一美，大隅典子，上條竜太郎
  • Organizer: 第46回日本口腔科学会関東地方部会
  • Place of Presentation: 埼玉医科大学（川越）
• [Presentation] 成体マウス毛包内の神経堤由来細胞による象牙芽細胞分化誘導 (2012)
  • Author(s): 森澤絵里，須澤徹夫，宮内知彦，鈴木航，馬場一美，上條竜太郎
  • Organizer: 第54回歯科基礎医学会学術大会・総会
  • Place of Presentation: 奥羽大学（郡山）
• [Presentation] 口腔粘膜上皮のケラチノサイトの一部は神経堤に由来する (2012)
  • Author(s): 小野美樹，高見正道，須澤徹夫，綿引淳一，榎本明子，南保友樹，田口智博，市川雄大，野瀬佳奈，上條竜太郎，槇宏太郎
  • Organizer: 第71回日本矯正歯科学会大会
  • Place of Presentation: 盛岡市アイスアリーナ(盛岡)
• [Presentation] LPS Inhibits Ectopic Bone Formation Induced by BMP-2 plus TGF-β1 in Mice (2012)
  • Author(s): Matsumoto A, Takami M, Miyamoto A, Tachi K, Suzawa T, Baba K, Kamijo R.
  • Organizer: The ASBMR 2012 Annual Meeting
  • Place of Presentation: Minneapolis, MN, USA
• [Book] The New Frontiers in Research for Oral Cancer (2012)
  • Author(s): Tachikawa T.
  • Total Pages: 209
  • Publisher: MARUZEN PLANET
• [Remarks] 昭和大学歯学部口腔生化学講座
  • URL: http://www10.showa-u.ac.jp/~0rqlbio/
• [Remarks] http://www10.showa-u.ac.jp/~oralbio/
• [Remarks] 昭和大学歯学部口腔生化学講座
  • URL: http://www10.showa-u.ac.jp/~oralbio/