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Molecular hydrogen suppresses Porphyromonas gingivalis lipopolysaccharide-induced increases in interleukin-1 alpha and interleukin-6 secretion in human gingival cells

Abstract

Periodontitis is defined as a multifactorial polymicrobial infection accompanied by inflammatory reactions. Porphyromonas gingivalis (Pg) is known as a major pathogen in the initiation and progression of periodontitis, and a major virulence factor is Pg lipopolysaccharide (LPS). Molecular hydrogen (H2) has been reported to act as a gaseous antioxidant, which suppresses periodontitis progression by decreasing gingival oxidative stress. However, no human periodontitis model has examined the anti-inflammatory effects of H2. In this study, we examined the effects of H2 on Pg LPS-induced secretion of 8 types of inflammation markers in a human periodontitis model using human gingival cells with enzyme-linked immunosorbent assays. Our results demonstrated that Pg LPS increased interleukin (IL) 1 alpha (IL-1α) and IL-6 secretion, but H2 significantly suppressed the secretion of both cytokines without cytotoxicity. H2 can suppress the production of IL-1α and IL-6, which are identified as cytokines involved in inflammatory reactions in periodontal disease. Thus, H2 may provide therapeutic applications for periodontitis.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was partly supported by Fukuyama city support project for research & development and market development [Grant Number #236, 2018] and JSPS KAKENHI Grant Number 20K11627.

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YS designed the study. NY, DM, and AM performed the experiments and analyzed the data. YS wrote the manuscript. All authors approved the final version of the manuscript.

Corresponding author

Correspondence to Yasukazu Saitoh.

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YS and NY declare that they have no conflict of interest. DM and AM are employees of Hiroshima Kasei, Ltd.

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Supplementary Information

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11010_2021_4262_MOESM1_ESM.pptx

Supplementary file1 Supplementary Fig. 1 Time-dependent change in H2 concentration in cell culture condition. Culture medium containing dissolved H2 was produced, and incubated at 37 °C in a humidified atmosphere of 95% air and 5% CO2 as same condition of the cell culture. Thereafter, the time-dependent change in H2 concentration was measured for 3 h. Results are expressed as mean ± SD (n = 3). Significant differences from the control, **p < 0.01. Supplementary Fig. 2 The effect of Pg LPS treatment on intracellular oxidative stress level. Intracellular oxidative stress level was determined using a fluorescent assay performed using 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate, di (acetoxymethyl ester) (CDCFH-DA). Briefly, cells were rinsed with cell culture medium, then incubated for 30 min at 37 °C in the medium containing 10 µM CDCFH-DA. After Pg LPS treatment and incubation for 24 h, cells were washed with medium and the oxidative stress-derived fluorescence was measured using a fluorescence microscope reader with excitation and emission wavelengths of 485 nm and 530 nm, respectively. Then, cell number was counted in each well. Results are expressed as mean ± SD (n = 5) (PPTX 46 kb)

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Saitoh, Y., Yonekura, N., Matsuoka, D. et al. Molecular hydrogen suppresses Porphyromonas gingivalis lipopolysaccharide-induced increases in interleukin-1 alpha and interleukin-6 secretion in human gingival cells. Mol Cell Biochem (2021). https://doi.org/10.1007/s11010-021-04262-7

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Keywords

  • Periodontitis
  • Molecular hydrogen
  • Human gingival cell
  • Porphyromonas gingivalis
  • Interleukin-1α
  • Interleukin-6