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The increase in large-scale production of magnetic nanoparticles (NP) associated with the incomplete comprehensive knowledge regarding the potential risks of their use on environmental and human health makes it necessary to study the biological effects of these particles on organisms at the cellular level. The aim of this study to examine the cellular effects on fibroblast lineage LA-9 after exposure to mixed iron oxide NP (Fe 3 O 4  NP). The following analyses were performed: field emission gun–scanning electron microscopy (SEM-FEG), dynamic light scattering (DLS), zeta potential, ultraviolet/visible region spectroscopy (UV/VIS), and attenuated total reactance-Fourier transform infrared (ATR-FTIR) spectroscopy analyses for characterization of the NP. The assays included cell viability, morphology, clonogenic potential, oxidative stress as measurement of reactive oxygen species (ROS) and nitric oxide (NO) levels, cytokines quantification interleukin 6 (IL-6) and tumor necrosis factor (TNF), NP uptake, and cell death. The size of Fe 3 O 4  NP was 26.3 nm when evaluated in water through DLS. Fe 3 O 4  NP did not reduce fibroblast cell viability until the highest concentration tested (250 µg/ml), which showed a decrease in clonogenic potential as well as small morphological changes after exposure for 48 and 72 hr. The NP concentration of 250 µg/ml induced enhanced ROS and NO production after 24 hr treatment. The uptake assay exhibited time-dependent Fe 3 O 4  NP internalization at all concentrations tested with no significant cell death. Hence, exposure of fibroblasts to Fe 3 O 4  NP-induced oxidative stress but not reduced cell viability or death. However, the decrease in the clonogenic potential at the highest concentration demonstrates cytotoxic effects attributed to Fe 3 O 4  NP which occurred on the 7 th  day after exposure.

Toxicological effects of the mixed iron oxide nanoparticle (Fe 3 O 4 NP) on murine fibroblasts LA-9