Abstract
Hydrogen peroxide is a normal by-product of cellular metabolism that in higher concentrations can cause oxidative stress. Reactive oxygen species impair the physiological functions of retinal pigment epithelial (RPE) cells, which are known as one major cause of ocular pathologies. Most studies investigating the influence of H2O2 on cells in culture but H2O2 concentrations are not sustained in culture medium. Continuous generation using glucose oxidase (GOx) system allows application of relevant low H2O2 concentrations over physiologically relevant times periods (up to 24 h). Recent findings suggest that bolus and GOx treatments can lead to different cellular response, thus warranting a quantitative comparison between the two approaches. When added as a pulse H2O2 is rapidly depleted. Continuous generation of H2O2 produces different behavior in function of GOx activities. Cytotoxicity analyses show that cells can tolerate short exposure to high H2O2 doses delivered as a pulse but are susceptible to lower continuous doses. Application of hydrogen peroxide causes a concentration-dependent decrease in the intracellular glutathione (GSH) content that was accompanied by a matching decrease in the glutathione peroxide activity and reducing power (FRAP).
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