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    Differential Sensitivity of Two Leukemia Cell Lines towards Two Major Gas Plasma Products Hydrogen Peroxide and Hypochlorous Acid
    (Basel : MDPI, 2022) Singer, Debora; Miebach, Lea; Bekeschus, Sander
    Oxidative stress has major implications for health and disease. At the same time, the term collectively describes the reactions to different types of reactive oxygen species (ROS) and oxidants, including hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). However, how both compare in terms of cytotoxicity and mechanism of action is less known. Using two leukemia cell lines, Jurkat and THP-1, as model systems at similar cell concentrations, we found an 8-fold greater sensitivity of the former over the latter for H2O2 exposure. Unexpectantly, this was not the case with HOCl exposure. Jurkat cells were 2-fold more resistant to HOCl-induced cytotoxicity than THP-1 cells. In each cell type, the relatively more toxic oxidant also induced activation of caspases 3 and 7 at earlier time points, as time-lapse fluorescence microscopy revealed. The effects observed did not markedly correlate with changes in intracellular GSH and GSSG levels. In addition, siRNA-mediated knockdown of the Nrf2 target HMOX-1 encoding for HO-1 protein and the growth and survival factor IL-8 revealed Jurkat cells to become more sensitive to HOCl, while HO-1 and IL-8 siRNA-mediated knockdown in THP-1 cells produced greater sensitivity towards H2O2. siRNA-mediated knockdown of catalase increased oxidant sensitivity only negligibly. Collectively, the data suggest striking HOCl-resistance of Jurkat and H2O2 resistance of THP-1 cells, showing similar protective roles of HO-1 and IL-8, while caspase activation kinetics differ.
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    Cold Atmospheric Pressure Plasma Is Effective against P. gingivalis (HW24D-1) Mature Biofilms and Non-Genotoxic to Oral Cells
    (Basel : MDPI, 2022) de Morais Gouvêa Lima, Gabriela; Carta, Celina Faig Lima; Borges, Aline Chiodi; Nishime, Thalita Mayumi Castaldelli; da Silva, Cézar Augusto Villela; Caliari, Marcelo Vidigal; Mayer, Marcia Pinto Alves; Kostov, Konstantin Georgiev; Koga-Ito, Cristiane Yumi
    The effects of helium cold atmospheric pressure plasma (He-CAPP) jet on Porphyromonas gingivalis (HW24D-1) biofilm, on human gingival fibroblasts (HGF) and human gingival keratinocytes (OBA-9) were assessed. Standardized suspension of P. gingivalis was obtained, and biofilms were grown anaerobically for 48 h. After exposition to He-CAPP, the biofilm viability was evaluated by XTT assay. HGF were grown at 37 °C, in an CO2 chamber in DMEM, while OBA-9 cells were cultured in keratinocyte serum-free medium. After 24 h, plates were exposed to He-CAPP for 1 to 7 min. Plasma was generated using a commercial AC power supply with amplitude modulated signal (voltage amplitude of 20 kVp-p, frequency of 31.0 kHz and duty cycle of 22%). The corresponding discharge power was 0.6W at He flow rate of 1 L/min. DNA damage was accessed by static cytometry. Data were analyzed by GraphPad Prism (p < 0.05). Significant reductions in P. gingivalis viability in relation to non-treated groups were detected (p < 0.0001), directly proportional to exposure time. Treated groups were slightly aneuploid after 5- and 7-min treatment in HGF, and for 3 min in OBA-9 cells, with 1.2 DNA index mean. Helium cold atmospheric pressure plasma jet showed inhibitory effect on P. gingivalis mature biofilm and was not genotoxic for epithelial gingival cells and human oral fibroblasts.