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Bioglasses have been used throughout the past century as a biomaterial in the bone regeneration field. However, recent studies have attempted to use them as a therapeutic material as well, mainly in the treatment of osteosarcomas. The most widely recognized bioglass is the 45S5 Bioglass, invented by Larry Hench et al., which presents higher bioactivity. A possible application of this bioglass in the treatment of osteosarcomas can be accomplished by adding specific ions, such as iron, that will allow the use of magnetic hyperthermia and Fenton reaction as therapeutic mechanisms. In this study, a 45S5 Bioglass containing 10%mol of Fe2O3 was produced using the melt-quenching method. A group of samples was prepared by changing the overall ball milling time, from 1 h up to 48 h, to analyze the effects of iron in the bioactive glass matrix and evaluate the influence of particle size on their physical and biological properties. The studied bioglasses showed no evidence of changes in the amorphous structural nature compared to the 45S5 Bioglass. The data of the impedance spectroscopy study revealed that the addition of Fe2O3 can increase the standard rate constant of the Electro-Fenton reaction, with the sample milled for 12 h showing the most promising results. The reduction in the particle size influenced the cytotoxicity and the bioactivity. The samples with lower particle sizes showed a higher level of cytotoxicity.