Ozone ultrafine bubble water induces the cellular signaling involved in oxidative stress responses in human periodontal ligament fibroblasts

Anongwee Leewananthawet,Shinichi Arakawa,Tokuju Okano,Ryo Daitoku Kinoshita,Hiroshi Ashida,Yuichi Izumi,Toshihiko Suzuki

doi:10.1080/14686996.2019.1614980

Anongwee Leewananthawet, Shinichi Arakawa + Show 5 more

Open Access

https://doi.org/10.1080/14686996.2019.1614980

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Abstract

ABSTRACT 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.

Highlights

Ozone (O3) is a strong oxidizing agent that is widely used as an antiseptic in the food industry and dental fields
To investigate the cytotoxicity of ozone ultrafine bubble water (OUFBW) at the cellular level, we used human primary periodontal ligament fibroblasts, since these cells play an important role in the maintenance and regeneration of periodontal tissues
The cells were exposed to OUFBW for 1 or 10 min, because the treatment time with OUFBW is assumed to be less than 10 min in clinical use, and the cells were further incubated for 3 h in the medium

Summary

Introduction

Ozone (O3) is a strong oxidizing agent that is widely used as an antiseptic in the food industry and dental fields. Regarding practical application of its antimicrobial activity, ozone has been used for water purification and food preservation, and more recently, as a disinfectant in the treatment of dental caries, endodontic lesions and periodontal diseases [1,2,3,4,5,6,7]. The strong bactericidal efficacy of ozone results from its strong oxidizing activity. Ozone is known to induce oxidative stress in cells via Supplemental data for this article can be accessed here. Note that inhalation of high concentrations of gaseous ozone has been suggested as a possible risk factor in the development of asthma and allergic airway inflammation [8,9]

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