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An in vitro senescence model of gingival epithelial cell induced by hydrogen peroxide treatment

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Abstract

Gingival tissue shows progressive changes with aging and an in vitro model of gingival tissue could be useful in understanding age-associated oral diseases. The present study aims to establish a hydrogen peroxide (H2O2) treatment model to induce aging in human gingival epithelial cells. In addition, fisetin, a flavonoid component studied for the anti-aging property is used to examine if it could reverse the induced senescence. Primary human gingival epithelial progenitor (HGEPp) cells were cultured and treated with different concentrations of H2O2. A cell vitality and morphology, senescence-associated beta-galactosidase (SA-β-gal) staining, mRNA and protein expression analysis of known senescence markers p16, p21, and p53, and cell cycle assay were performed. The cells showed dose-dependent changes in vitality and morphology, SA-β-gal staining, relative mRNA and protein expression, and cell cycle assay after H2O2 treatment. Based on these results, 400 μM H2O2 was considered as an optimal concentration to induce senescence. Treatment of senescence-induced cells with fisetin downregulated all the senescence markers used in this study. In conclusion, a senescence model of gingival epithelial cells induced by hydrogen peroxide treatment was established which could be employed to study age-related periodontal diseases.

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Funding

Grant was received to conduct this study.

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Authors and Affiliations

  1. Division of Periodontology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, 061-0293, Japan

    Sarita Giri & Yasushi Furuichi

  2. Division of Biochemistry, Department of Oral Biology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan

    Ayuko Takada

  3. Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan

    Durga Paudel, Koki Yoshida & Yoshihiro Abiko

  4. Department of Oral Disease Research, National Center for Geriatrics and Gerontology, Obu, Japan

    Masae Furukawa & Kenji Matsushita

  5. Research Institute of Cancer Prevention, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan

    Yasuhiro Kuramitsu

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  1. Sarita Giri
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Contributions

YF conceptualized the idea. YF, AT, KM and MF designed the study. SG, AT, KY and YK conducted the experiments. SG, DP, YA and YF analyzed and interpreted the data. SG wrote the initial draft of the manuscript. YA and YF contributed in the critical revision of the manuscript. All authors agreed to the final version of the manuscript.

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Correspondence to Yasushi Furuichi.

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Giri, S., Takada, A., Paudel, D. et al. An in vitro senescence model of gingival epithelial cell induced by hydrogen peroxide treatment. Odontology 110, 44–53 (2022). https://doi.org/10.1007/s10266-021-00630-3

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  • DOI: https://doi.org/10.1007/s10266-021-00630-3

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