1. Safety 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 modelIza Rozman, Álvaro Gallo-Cordova, María del Puerto Morales, Marco A. Morales Ovalle, Gerardo F. Goya, Katja Kološa, Domen Hočevar, Bojana Žegura, Alja Štern, 2026, original scientific article Abstract: Given 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.
Keywords: DNA damage, genotoxicity, HepG2 spheroids, magnetic ferrite-based nanoparticles, ROS induction, safety assessment, toxicogenomics
Published in DiRROS: 27.01.2026; Views: 189; Downloads: 150
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2. Nano spray-dried particles of in-situ crosslinked alginate and their toxicological characterisationJaka Levanič, Ilja Gasan Osojnik Črnivec, Iza Rozman, Mihaela Skrt, Alja Štern, Bojana Žegura, Nataša Poklar Ulrih, 2024, original scientific article Abstract: The feasibility and technical capacity for producing crosslinked sub-micron gels with a nano spray-dryer were studied with variable pH systems incorporating alginate, pectin, and pullulan. The obtained powders were characterized for their morphology, particle size distribution, and their toxicological safety profile using genotoxicity and cytotoxicity assays. Additionally, quercetin was added to the encapsulation system to study the potential of the system to encapsulate this material. The produced powders exhibited morphologies and particle size distributions characteristic for nano spray-drying. The addition of pullulan and pectin to the feed solutions resulted in a particle size increase, with crosslinked alginate particles having a mean value of 1.43 μm, while particles with added pectin and pullulan had a mean particle size of 1.70 and 1.75 μm, respectively. The inclusion of quercetin proved to be problematic with this encapsulation system. Extremely high degradation rates and extremely low encapsulation efficiencies were observed due to the alkaline pH (~10) of the system that is needed to keep the feed dispersion in a liquid state and prevent premature crosslinking of the alginate. Although pectin and pullulan provided some protection for quercetin in the alkaline dispersion, the absolute quercetin content in the final product remained very low, with a maximum achieved encapsulation efficiency of 2.06 %. The safety profile of most produced powders was favourable, as they did not exhibit any significant cytotoxic and genotoxic activity in the HepG2 cell line, except in the case of Alginate/Pullulan which showed a 43 % decrease in cell viability at 500 μg/mL. Samples where quercetin was added did not show any increased toxicological effect.
Keywords: alginate, polysaccharides, crosslinking, nano spray drying, toxicological evaluation
Published in DiRROS: 08.05.2025; Views: 853; Downloads: 526
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3. Genotoxicity and heating performance of VxFe3-xO4 nanoparticles in health applicationsBeatriz Sanz-Sagué, Amaia Sáenz-Hernández, Bojana Žegura, Alja Štern, Katja Kološa, Iza Rozman, 2024, original scientific article Abstract: The applications of magnetic nanoparticles (MNPs) as biocatalysts in different biomedical areas have been evolved very recently. One of the main challenges in this field is to design affective MNPs surfaces with catalytically active atomic centres, while producing minimal toxicological side effects on the hosting cell or tissues. MNPs of vanadium spinel ferrite (VFe2O4) are a promising material for mimicking the action of natural enzymes in degrading harmful substrates due to the presence of active V5+ centres. However, the toxicity of this material has not been yet studied in detail enough to grant biomedical safety. In this work, we have extensively measured the structural, compositional, and magnetic properties of a series of VxFe3-xO4 spinel ferrite MNPs to assess the surface composition and oxidation state of V atoms, and also performed systematic and extensive in vitro cytotoxicity and genotoxicity testing required to assess their safety in potential clinical applications. We could establish the presence of V5+ at the particle surface even in water-based colloidal samples at pH 7, as well as different amounts of V2+ and V3+ substitution at the A and B sites of the spinel structure. All samples showed large heating efficiency with Specific Loss Power values up to 400 W/g (H0 = 30 kA/m; f = 700 kHz). Samples analysed for safety in human hepatocellular carcinoma (HepG2) cell line with up to 24h of exposure showed that these MNPs did not induce major genomic abnormalities such as micronuclei, nuclear buds, or nucleoplasmic bridges (MNIs, NBUDs, and NPBs), nor did they cause DNA double-strand breaks (DSBs) or aneugenic effects—types of damage considered most harmful to cellular genetic material. The present study is an essential step towards the use of these type of nanomaterials in any biomedical or clinical application.
Keywords: magnetic nanoparticles, vanadium ferrite, cytotoxicity, genotoxicity, specific power absorption, cell viability
Published in DiRROS: 23.05.2024; Views: 1245; Downloads: 907
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