The purpose of our study was to investigate basic mechanisms of hydrogen (H₂) gas – induced cardioprotection during oxidative stress in a cardiomyocytes cell culture model. H₂ gas has been demonstrated to reduce the hydroxyl radical (•OH), the most cytotoxic of reactive oxygen species (ROS), and effectively protected cells. Here, we developed a new model for evaluating that H₂ gas has the potential to act as antioxidant on H₂O₂-induced death of H9c2 cell. Treatment with H₂O₂ (500µM) induced death of H9c2 cells. However, co-treatment with H₂ gas increases viability of cells. These results suggest that method is a useful alternative animal model to clarify the mechanism which H₂ gas protects H9c2 cardiomyocytes from oxidative stress.

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A cell recovery method, cells are exposed to chemicals and then cultured in chemical-free medium for a specific period, and toxicity is evaluated based on the recovery level of the cells. Cells exposed to strongly toxic chemicals do no not recover, and the cytotoxicity of some chemicals may be enhanced by chemical-free medium. In this study, to investigate the application of this method for ES-D3 cell differentiation, in addition to cell proliferation, we evaluated 4 types of monomer used for dental composite resin and adhesive, and observed that cells exposed to Bis-GMA did not recover compared to those exposed to the other monomers. The cell differentiation levels were the same as those observed under the conventional conditions, showing that further improvement of the cell recovery test method is necessary.

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