Leewananthawet Anongwee, Arakawa Shinichi, Okano Tokuju, Daitoku Kinoshita Ryo, Ashida Hiroshi, Izumi Yuichi, Suzuki Toshihiko
Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Sci Technol Adv Mater. 2019 Jun 13;20(1):589-598. doi: 10.1080/14686996.2019.1614980. eCollection 2019.
Periodontitis is a chronic inflammatory disease caused by oral microorganisms in the subgingival biofilm. Stable aqueous ozone ultrafine bubble water (OUFBW) has recently begun to be used as an antiseptic in the treatment of periodontitis. The effectiveness of OUFBW is thought to depend on the bactericidal actions of dissolved ozone exerted via its oxidizing effect. On the other hand, the effects of ozone on the periodontal tissues are largely unknown. In this paper we examined the cellular responses after OUFBW treatment. Human primary periodontal ligament fibroblasts (hPDLFs) or Ca9-22 human gingival epithelial cells were treated with OUFBW or UV-inactivated OUFBW. The production of reactive oxygen species (ROS), the activation of mitogen-activated protein kinase (MAPK) and the nuclear factor-kappa B (NF-κB) activation were analyzed. The transcript profiles of hPDLFs after OUFBW treatment were also analyzed by RNA sequencing (RNA-seq). Our results showed that OUFBW induces oxidative stress by generating ROS, which, in turn, activated the MAPK pathway. OUFBW triggered activation of c-Fos, a major component of the transcription factor activator protein 1 (AP-1), and also nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2), which possessed a high sensitivity to oxidative stress. The results of RNA-seq analysis revealed that the numerous genes involved in oxidative stress responses or MAPK signaling pathway were up-regulated after OUFBW treatment. Investigation of the signaling pathways activated by OUFBW highlights another aspect of the biological roles of OUFBW, in addition to its bactericidal activity, in the treatment of periodontitis.
牙周炎是一种由龈下生物膜中的口腔微生物引起的慢性炎症性疾病。稳定的水性臭氧超细气泡水（OUFBW）最近开始被用作治疗牙周炎的抗菌剂。OUFBW的有效性被认为取决于溶解臭氧通过其氧化作用发挥的杀菌作用。另一方面，臭氧对牙周组织的影响在很大程度上尚不清楚。在本文中，我们研究了OUFBW处理后的细胞反应。用人原代牙周膜成纤维细胞（hPDLFs）或Ca9-22人牙龈上皮细胞用OUFBW或紫外线灭活的OUFBW处理。分析了活性氧（ROS）的产生、丝裂原活化蛋白激酶（MAPK）的激活和核因子-κB（NF-κB）的激活。还通过RNA测序（RNA-seq）分析了OUFBW处理后hPDLFs的转录谱。我们的结果表明，OUFBW通过产生活性氧诱导氧化应激，进而激活MAPK途径。OUFBW触发了转录因子激活蛋白1（AP-1）的主要成分c-Fos以及对氧化应激具有高敏感性的核因子红细胞2（NF-E2）相关因子2（Nrf2）的激活。RNA-seq分析结果显示，OUFBW处理后，许多参与氧化应激反应或MAPK信号通路的基因上调。对OUFBW激活的信号通路的研究突出了OUFBW在治疗牙周炎中的生物学作用的另一个方面，除了其杀菌活性之外。
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