DNA Damage Induction After Exposure to Ferrite Nanoparticles for biomedical applications (MxFe3-xO4, M = Fe, Zn, Mn) in an Advanced 3D Human Hepatic in vitro ModelRozman, Iza (Avtor) Štern, Alja (Avtor) Žegura, Bojana (Avtor) Gallo-Cordova, Álvaro (Avtor) Puerto Morales, María del (Avtor) Hočevar, Domen (Avtor) Goya, Gerardo F. (Avtor) TEMferrite-based nanoparticlesHepG2 spheroidsDNA damagecomet assayNanosized spinel-type ferrites have gained recognition as a unique class of engineered nanomaterials with promising applications, but their safety profiles remain insufficiently explored. Although iron (Fe), zinc (Zn), and manganese (Mn) are biologically relevant elements, the use of Zn- and Mn-containing ferrite nanoparticles in biomedical contexts demands careful (geno)toxicity evaluation. In this study, three ferrite nanoparticles – γFe2O3 (FeNPs), Zn0.7Fe2.3O4 (ZnNPs), and Mn0.4Fe2.6O4 (MnNPs) – synthesised through a microwave-assisted polyol route, functionalized with citric acid to improve colloidal stability, were evaluated for their potential (geno)toxic effects in an advanced in vitro 3D cell model, HepG2 spheroids. DNA damage induction was studied using the alkaline comet assay. After 24 and 96 hours of exposure, spheroids were dissociated into a single-cell suspension by collagenase treatment. Each spheroid was incubated in 0.25% trypsin–EDTA for 3 minutes and then gently dissociated into a single-cell suspension using cut pipette tips. After this the comet assay was conducted under conditions described in the Supplement material (Table 1). The cell suspension was combined with 1% low-melting-point (LMP) agarose and layered onto fully frosted slides pre-coated with 1% normal-melting-point (NMP) agarose. Slides were lysed, the nucleoids unwound and electrophoresis preformed, followed by neutralisation of the gells.Zenodo20252026-02-13 03:50:44Neznano27561UDK: 577.2DOI: 10.5281/zenodo.17896231COBISS_ID: 266870019sl