Study of the inhibitory effects of polyphenol loaded titania nanotubes on bacterial growth and biofilm formation

• Project/Area Number: 19K10132
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57030:Conservative dentistry-related
• Research Institution: The University of Tokushima
• Principal Investigator: OZAKI Kazumi 徳島大学, 大学院医歯薬学研究部(歯学域), 教授 (90214121)
• Co-Investigator(Kenkyū-buntansha): 湯本 浩通 徳島大学, 大学院医歯薬学研究部(歯学域), 教授 (60284303) ; 細川 義隆 徳島大学, 病院, 講師 (90346601) ; 細川 育子 徳島大学, 大学院医歯薬学研究部(歯学域), 助教 (50707908)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: バイオフィルム / 二酸化チタン / チタニアナノチューブ / ポリフェノール / 光触媒

Outline of Research at the Start

TNT-PolyP mixture（以下，合剤）として期待する効果が最大限に発揮される至適な形状のTiO2 nanotubes（TNT）を形態学的解析によって確認しつつ合成する。一方，TNTに保持するPolyP（EGCGとCCM）の至適濃度や混合比を，細菌学的解析等を通して決定する。そして，合剤の口腔バイオフィルムの初期形成阻止や成長抑制の効果を検証するとともに，齲蝕誘発モデルで形成させたバイオフィルム内の菌体内外多糖および歯質の超微構造に合剤が与える影響を解析する。さらに，歯根部試料を調製し，歯根表面の耐酸性や再石灰化能に及ぼす合剤の効果を検証する。これらの検証を行った上，臨床応用に向けた具体的な技術指針を確立する。

Outline of Final Research Achievements

The aim of this study was to evaluate the inhibitory effect of polyphenol loaded titania nanotubes (TNT-PolyP mixture) on bacterial growth and biofilm formation. After preparing a TNT-PolyP mixture, the Streptococcus mutans UA159 strain was cultured for 12 hours using a sucrose-added BHI medium and apatite pellets supplemented with the TNT-PolyP mixture.
As a result, a certain degree of antibacterial effect and biofilm formation inhibitory effect were observed. TEM observation revealed that the TNT-PolyP mixture suppresses the biofilm formation of S.mutans. In addition, it showed a tendency to reduce the roughening of the surface of apatite pellets, that is, the decalcification effect. From the above, it was shown that the TNT-PolyP mixture prepared in this study may be useful for suppressing the onset of caries.

Academic Significance and Societal Importance of the Research Achievements

う蝕や歯周病の発症要因となるだけでなく，誤嚥性肺炎など死に至る疾患の原因ともいわれる口腔バイオフィルムの駆逐技術を開発することは，歯科臨床上非常に意義深い。
とくに口腔衛生管理の手段として患者に指導するブラッシングなど口腔バイオフィルムの機械的除去に加え，本研究で調製した合剤のような新たなコンセプトに基づく口腔バイオフィルム形成制御技術が応用されることで，機能障害の有無を問わず高度なブラッシングを要求しがたい高齢者や要介護者の口腔衛生管理の質が向上し，根面う蝕の発症予防や進行抑制に貢献すると考えられる。

Research Products

• [Journal Article] Nobiletin inhibits inflammatory reaction in interleukin-1β-stimulated human periodontal ligament cells. (2021)
  • Author(s): Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Takashi Matsuo.
  • Journal Title: Pharmaceutics Volume: 13 Issue: 5 Pages: 667-667
  • DOI: 10.3390/pharmaceutics13050667
  • NAID: 120007188425
  • Peer Reviewed / Open Access
• [Journal Article] Nobiletin Decreases Inflammatory Mediator Expression in Tumor Necrosis Factor-Stimulated Human Periodontal Ligament Cells., (2021)
  • Author(s): Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki
  • Journal Title: Mediators of Inflammation Volume: 2021 Pages: 1-7
  • DOI: 10.1155/2021/5535844
  • NAID: 120007185165
  • Peer Reviewed / Open Access
• [Journal Article] Conversion from epithelial to partial-EMT phenotype by Fusobacterium nucleatum infection promotes invasion of oral cancer cells. (2021)
  • Author(s): Wenhua Shao, Natsumi Fujiwara, Yashiro Mouri, Satoru Kisoda, Kayo Yoshida, Kaya Yoshida, Hiromichi Yumoto, Kazumi Ozaki, Naozumi Ishimaru, Yasusei Kudo
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 14943-14943
  • DOI: 10.1038/s41598-021-94384-1
  • NAID: 120007168795
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular vesicles of P. gingivalis-infected macrophages induce lung injury. (2021)
  • Author(s): Kayo Yoshida, Kaya Yoshida, Natsumi Fujiwara, Mariko Seyama, Kisho Ono, Hotaka Kawai, Jiajie Guo, Ziyi Wang, Yao Weng, Yaqiong Yu, Yoko Uchida-Fukuhara, Mika Ikegame, Akira Sasaki, Hitoshi Nagatsuka, Hiroshi Kamioka, Hirohiko Okamura, Kazumi Ozaki
  • Journal Title: Biochim Biophys Acta - Molecular Basis of Disease Volume: 1867 Issue: 11 Pages: 166236-166236
  • DOI: 10.1016/j.bbadis.2021.166236
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Suppressive effects of 2-Methacryloyloxyethyl phosphorylcholine (MPC)-polymer on the adherence of Candida albicans and MRSA to acrylic denture resin. (2020)
  • Author(s): N. Fujiwara, K. Murakami, K. Yoshida, S. Sakurai, Y. Kudoh, K. Ozaki, K. Hirota, H. Fujii, M. Suzuki, Y. Miyake, H. Yumoto
  • Journal Title: Heliyon Volume: 6 Issue: 6 Pages: e04211-e04211
  • DOI: 10.1016/j.heliyon.2020.e04211
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular Vesicles in Periodontal Medicine: (2020)
  • Author(s): Yoshida K., Yoshida K., Seyama M., Ozaki K., Okamura H.
  • Journal Title: Journal of Oral Health and Biosciences Volume: 33 Issue: 1 Pages: 15-23
  • DOI: 10.20738/johb.33.1_15
  • NAID: 130007854091
  • ISSN: 2188-7888, 2189-6682
  • Peer Reviewed
• [Journal Article] Carnosic acid inhibits inflammatory cytokines production in human periodontal ligament cells. (2020)
  • Author(s): Hosokawa I., Hosokawa Y., Ozaki K., Matsuo T.
  • Journal Title: Immunopharmacology and Immunotoxicology Volume: 42 Issue: 4 Pages: 373-378
  • DOI: 10.1080/08923973.2020.1782427
  • Peer Reviewed
• [Journal Article] Involvement of Fusobacterium Species in Oral Cancer Progression：A Literature Review Including Other Types of Cancer. (2020)
  • Author(s): Fujiwara N., Kitamura N., Yoshida K., Yamamoto T.,Ozaki K., Kudo Y.
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 17 Pages: 6207-6216
  • DOI: 10.3390/ijms21176207
  • Peer Reviewed / Open Access
• [Journal Article] Outer membrane vesicles of Porphyromonas gingivalis attenuate insulin sensitivity by delivering gingipains to the liver. (2020)
  • Author(s): Seyama M, Yoshida K, Yoshida K, Fujiwara N, Ono K, Eguchi T, Kawai H, Guo J, Weng Y, Haoze Y, Uchibe K, Ikegame M, Sasaki A, Nagatsuka H, Okamoto K, Okamura H, Ozaki K.
  • Journal Title: Biochim Biophys Acta Mol Basis Dis. Volume: 1866(6) Issue: 6 Pages: 165731-165731
  • DOI: 10.1016/j.bbadis.2020.165731
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Carnosic Acid Inhibits CXCR3 Ligands Production in IL-27-Stimulated Human Oral Epithelial Cells (2019)
  • Author(s): Hosokawa Ikuko、Hosokawa Yoshitaka、Ozaki Kazumi、Matsuo Takashi
  • Journal Title: Inflammation Volume: 印刷中 Issue: 4 Pages: 1311-1316
  • DOI: 10.1007/s10753-019-00991-6
  • Peer Reviewed / Open Access
• [Journal Article] Sudachitin Inhibits Matrix Metalloproteinase-1 and -3 Production in Tumor Necrosis Factor-α-Stimulated Human Periodontal Ligament Cells. (2019)
  • Author(s): Yoshitaka Hosokawa, Ikuko Hosokawa, Kazumi Ozaki, Takashi Matsuo
  • Journal Title: Inflammation Volume: 42 Issue: 4 Pages: 1456-1462
  • DOI: 10.1007/s10753-019-01007-z
  • Peer Reviewed / Open Access
• [Presentation] Nobiletinはヒト歯根膜由来細胞のIL-1β誘導炎症性メディエーター産生を抑制する (2021)
  • Author(s): 細川 義隆, 細川 育子, 尾崎 和美
  • Organizer: 日本歯科保存学会2021年度春季学術大会(第154回)
• [Presentation] 歯周病原菌感染マクロファージの細胞外小胞が肺炎を誘導する可能性の検証 (2020)
  • Author(s): 吉田 佳世，吉田 賀弥，瀬山 真莉子，尾崎 和美
  • Organizer: 第63回春季日本歯周病学会学術大会(オンライン学術大会)
• [Presentation] 歯周病原菌感染マクロファージの細胞外小胞が肺に線維化を誘導する可能性の検証 (2020)
  • Author(s): 吉田 佳世，吉田 賀弥，瀬山 真莉子，尾崎 和美
  • Organizer: 第63回秋季日本歯周病学会学術大会(オンライン学術大会)
• [Presentation] 最終糖化産物はヒト口腔上皮細胞のリポカイン2発現を増加する (2019)
  • Author(s): 木戸 理恵，廣島 佑香，生田 貴久，吉田 賀弥，稲垣 裕司，成石 浩司，尾崎 和美，木戸 淳一，湯本 浩通
  • Organizer: 第62回春季日本歯周病学会学術大会
• [Presentation] 感染マクロファージの膜小胞は歯周病関連疾患発症に関与するか？ (2019)
  • Author(s): 吉田 賀弥，吉田 佳世，藤原 奈津美，尾崎 和美，岡村 裕彦
  • Organizer: 第38回 分子病理学研究会 淡路島シンポジウム
• [Presentation] Fusobacterium nucleatumが与える口腔癌細胞への影響 (2019)
  • Author(s): 藤原 奈津美, Shao Wenhua,吉田 賀弥,新垣 理恵子, 尾崎 和美,石丸 直澄, 工藤 保誠
  • Organizer: 第38回 分子病理学研究会 淡路島シンポジウム