000168612 001__ 168612 000168612 005__ 20260211123813.0 000168612 0247_ $$2doi$$a10.1016/j.biopha.2025.118950 000168612 0248_ $$2sideral$$a147999 000168612 037__ $$aART-2026-147999 000168612 041__ $$aeng 000168612 100__ $$aRozman, Iza 000168612 245__ $$aSafety of ferrite nanoparticles for biomedical applications: Cyto- and genotoxic effects of MxFe3-xO4 (M = Fe, Zn, Mn) in an Advanced 3D human hepatic in vitro model 000168612 260__ $$c2026 000168612 5060_ $$aAccess copy available to the general public$$fUnrestricted 000168612 5203_ $$aGiven the growing interest in nanosized spinel-type ferrite nanoparticles for biomedical applications and the limited information on their safety, this study aimed to assess their cellular and genotoxic effects in an in vitro 3D human hepatic cell model (HepG2 spheroids). Ferrite nanoparticles – γFe2O3 (FeNPs; 14 ± 4 nm), Zn0.7Fe2.3O4 (ZnNPs; 14 ± 5 nm), and Mn0.4Fe2.6O4 (MnNPs; 7 ± 2 nm) – were synthesised through a microwave-assisted polyol route, functionalized with citric acid, and characterised using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES), Transmission Electron Microscopy (TEM), X-ray Diffraction (XRD), and Fourier Transform Infrared Spectroscopy (FTIR). Nanoparticle uptake was analysed using TEM, cytotoxicity was measured with CellTiter-Glo®, and oxidative stress induction was assessed using the 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) and malondialdehyde (MDA) assay. Genotoxic effects were evaluated using the comet, γH2AX and p-H3 assays. Cellular stress responses were assessed using toxicogenomic analysis. Significant cytotoxicity of the tested nanoparticles (0.1–250 µg/mL) was observed; however, TEM analysis revealed limited penetration to the outermost cell layers of spheroids. Notably, only FeNPs induced ROS generation, while MDA levels remained unchanged in all tested samples. Low DNA damage was detected at 24 h, but a significant increase was observed at 96 h (5–50 µg/mL). No increase in γH2AX or p-H3 was found. No substantial alterations in DNA damage or oxidative stress-response gene expression were detected. Altogether, our findings suggest that the effects of ferrite nanoparticles are time- and composition-dependent, underlining the importance of further mechanistic and chronic exposure evaluations in 3D cell models. 000168612 536__ $$9info:eu-repo/grantAgreement/EC/HORIZON EUROPE/101079113/EU/TWINNING FOR EXCELLENCE TO STRATEGICALLY ADVANCE RESEARCH IN CARCINOGENESIS AND CANCER/CutCancer$$9info:eu-repo/grantAgreement/EC/H2020/101007629 /EU/Nanomaterials for Enzymatic Control of Oxidative Stress Toxicity and Free Radical Generation/NESTOR$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 101007629 -NESTOR 000168612 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttps://creativecommons.org/licenses/by/4.0/deed.es 000168612 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000168612 700__ $$aGallo-Cordova, Alvaro 000168612 700__ $$aPuerto Morales, María del 000168612 700__ $$aMorales Ovalle, Marco A. 000168612 700__ $$0(orcid)0000-0003-1558-9279$$aGoya, Gerardo F.$$uUniversidad de Zaragoza 000168612 700__ $$aKološa, Katja 000168612 700__ $$aHocevar, Domen 000168612 700__ $$aŽegura, Bojana 000168612 700__ $$aŠtern, Alja 000168612 7102_ $$12003$$2395$$aUniversidad de Zaragoza$$bDpto. Física Materia Condensa.$$cÁrea Física Materia Condensada 000168612 773__ $$g195 (2026), 118950 [15 pp.]$$pBiomed. pharmacother.$$tBIOMEDICINE & PHARMACOTHERAPY$$x0753-3322 000168612 8564_ $$s6450218$$uhttps://zaguan.unizar.es/record/168612/files/texto_completo.pdf$$yVersión publicada 000168612 8564_ $$s2443261$$uhttps://zaguan.unizar.es/record/168612/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000168612 909CO $$ooai:zaguan.unizar.es:168612$$particulos$$pdriver 000168612 951__ $$a2026-02-11-10:28:16 000168612 980__ $$aARTICLE