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Oxidative stress, largely mediated by reactive oxygen species (ROS), is a nearly ubiquitous component in complex biological processes such as aging and disease. Optimal in vitro methods used in elucidating disease mechanisms would deliver of low levels of hydrogen peroxide, emulating the in vivo pathological state, but current methods are limited by kinetic stability or accurate measurement of the dose administered. Here we present an in vitro platform that exploits anthraquinone catalysts for the photocatalytic production of hydrogen peroxide. This system can be dynamically tuned to provide constant generation of hydrogen peroxide at a desired physiologic rate over at least 14 days and is described using a kinetic model. Material characterization and stability is discussed along with a proof-of-concept in vitro study that assessed the viability of cells as they were oxidatively challenged over 24 h at different ROS generation rates.

Patients with cobalt chrome (CoCr) metal-on-metal (MOM) implants may be exposed to a wide size range of metallic nanoparticles as a result of wear. In this study we have characterised the biological responses of human fibroblasts to two types of synthetically derived CoCr particles [(a) from a tribometer (30 nm) and (b) thermal plasma technology (20, 35, and 80 nm)] in vitro, testing their dependence on nanoparticle size or the generation of oxygen free radicals, or both. Metal ions were released from the surface of nanoparticles, particularly from larger (80 nm) particles generated by thermal plasma technology. Exposure of fibroblasts to these nanoparticles triggered rapid (2 h) generation of reactive oxygen species (ROS) that could be eliminated by inhibition of NADPH oxidase, suggesting that it was mediated by phagocytosis of the particles. The exposure also caused a more prolonged, MitoQ sensitive production of ROS (24 h), suggesting involvement of mitochondria. Consequently, we recorded elevated levels of aneuploidy, chromosome clumping, fragmentation of mitochondria and damage to the cytoskeleton particularly to the microtubule network. Exposure to the nanoparticles resulted in misshapen nuclei, disruption of mature lamin B1 and increased nucleoplasmic bridges, which could be prevented by MitoQ. In addition, increased numbers of micronuclei were observed and these were only partly prevented by MitoQ, and the incidence of micronuclei and ion release from the nanoparticles were positively correlated with nanoparticle size, although the cytogenetic changes, modifications in nuclear shape and the amount of ROS were not. These results suggest that cells exhibit diverse mitochondrial ROS-dependent and independent responses to CoCr particles, and that nanoparticle size and the amount of metal ion released are influential.