Toxicology最新文献

{"title":"Effects of cigarette smoke extract on surfactant production in ATII-like cells, involvement of oxidative stress and autophagy","authors":"Maria Lisa Garavaglia ,&nbsp;Francesca Bodega ,&nbsp;Chiara Sironi ,&nbsp;Cristina Porta ,&nbsp;Isabella Dalle-Donne","doi":"10.1016/j.tox.2025.154262","DOIUrl":"10.1016/j.tox.2025.154262","url":null,"abstract":"<div><div>Surfactant plays an essential role in pulmonary physiology by reducing surface tension, preventing alveolar collapse, and regulating immune responses in the lung. The composition and function of surfactant can be modified by environmental pollutants. Among these, cigarette smoke is a leading cause of respiratory and cardiovascular diseases. However, the impact of cigarette smoke on surfactant composition and function is still poorly understood. This study evaluated the effects of cigarette smoke extract (CSE) on surfactant production in a differentiated cell model (A549/ATII-like). Specifically, we evaluated the effects of CSE on multilamellar bodies (MLBs) involved in surfactant synthesis and storage, lipid production and secretion, surface tension, and the expression of the surfactant proteins SP-B and SP-C. Since autophagy is involved in MLBs maturation, the expression of SQSTM1/p62 and of the phosphorylated protein pATG-16L1^S278 was evaluated to assess the effect of CSE exposure on the induction of the early stages of the autophagic process. Our results demonstrated that CSE exposure affects the expression of surfactant components in A549/ATII-like cells, inducing an increase in the surface tension at the air-water interface, possibly via oxidative stress and the alteration of autophagy.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154262"},"PeriodicalIF":4.6,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144840929","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A mechanistically-anchored and human stem cell-based in vitro test battery for assessing liver steatogenic potential of chemicals","authors":"Anouk Verhoeven,&nbsp;Alexandra Gatzios,&nbsp;Robim M. Rodrigues,&nbsp;Julen Sanz-Serrano,&nbsp;Joery De Kock,&nbsp;Mathieu Vinken ,&nbsp;Tamara Vanhaecke","doi":"10.1016/j.tox.2025.154263","DOIUrl":"10.1016/j.tox.2025.154263","url":null,"abstract":"<div><div>Fatty liver disease, which can result from various factors including chemical exposure, is an increasing clinical concern. A key event in its development is steatosis, referring to the accumulation of lipids within hepatocytes. To enable early detection of chemical-induced liver steatosis, we developed a mechanistically-anchored, human-relevant new approach methodology (NAM). This NAM consists of a 2-tiered <em>in vitro</em> test battery, aligned with the adverse outcome pathway (AOP) network for steatosis, and utilizes human skin-derived precursor cells differentiated into hepatic cells (hSKP-HPC), previously shown to be responsive to steatogenic triggers. In total, 6 well-known steatogenic compounds, including 3 pharmaceuticals (sodium valproate, tetracycline hydrochloride, amiodarone hydrochloride), a pesticide (cyproconazole), and 2 plasticizers (tricresyl phosphate and perfluorohexanesulfonic acid) alongside 2 non-steatogenic chemicals (minocycline hydrochloride and tartaric acid), were tested over a 72-hour period. Tier 1 evaluated transcriptional changes in key lipid metabolism pathways, and modulations were observed in nuclear receptors (<em>peroxisome proliferator-activated receptor),</em> fatty acid uptake <em>(fatty acid translocase)</em>, <em>de novo</em> lipogenesis <em>(diacylglycerol acyl transferase 2, fatty acid synthase and stearoyl-CoA desaturase 1)</em>, as well as in VLDL secretion <em>(apolipoprotein B100)</em>. Tier 2 assays assessed and confirmed downstream functional disruptions in fatty acid uptake and lipid accumulation as ultimate specific key events for steatogenic chemicals. Overall, this human stem cell-based NAM offers a promising tool for supporting early hazard identification of steatogenic chemicals across diverse sectors, bridging mechanistic insights to outcomes relevant to the initiation of fatty liver disease.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154263"},"PeriodicalIF":4.6,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144856427","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gadolinium impairs male steroidogenesis: In vivo and in vitro","authors":"Sara Falvo ,&nbsp;Giulia Grillo ,&nbsp;Massimo Venditti ,&nbsp;Gabriella Chieffi Baccari ,&nbsp;Maria Maddalena Di Fiore ,&nbsp;Tiziana Cappello ,&nbsp;Mariachiara Galati ,&nbsp;Maria Maisano ,&nbsp;Giuseppe Petito ,&nbsp;Rosalba Senese ,&nbsp;Alessandra Santillo","doi":"10.1016/j.tox.2025.154261","DOIUrl":"10.1016/j.tox.2025.154261","url":null,"abstract":"<div><div>The increasing use of gadolinium (Gd) in industrial and medical fields made it a hazardous environmental pollutant. Once ingested through water and/or food, Gd may potentially have toxic effects on all body districts. However, the effects of Gd on testicular function have been little explored. In the present study, adult male rats were allowed to drink GdCl₃ or Gd₂O₃ (10–20–40 mg/Kg b.w.) for 4 weeks. Following Gd treatment, a significant decrease in steroidogenic-related protein (StAR, 3β-HSD, 17β-HSD, and 5α-Red) expressions, T and DHT levels, and spermatozoa concentration were observed. To clarify the cellular mechanisms underlying Gd-induced damage, we exposed mouse Leydig (TM3) cells to increasing concentrations (5–1000 µM) of GdCl₃ or Gd₂O₃ for 24 h. The <em>in vitro</em> results showed a dose-dependent decrease in cell viability and confirmed that both Gd forms inhibited steroidogenesis-related protein expressions. Steroidogenesis is a multistep process taking place in mitochondria and endoplasmic reticulum, and Mitochondria-Associated Endoplasmic Reticulum Membranes (MAMs) play a key role. We found a decrease in MMP as well as in mitochondrial biogenesis (PGC1-α, NRF1, TFAM), fusion (MFN2), fission (DRP1) and MAMs (ATAD3, SOAT1, FACL4) marker expressions. The Gd-caused oxidative stress, as suggested by the increase in TBARS levels in GdCl₃- or Gd₂O₃-treated TM3 cells, increased autophagy and apoptosis by inhibiting the Akt pathway. In conclusion, our study highlights for the first time the adverse effects, and the underlying intracellular mechanisms, of orally administred Gd on the testicular function, laying the foundation for further research to understand its impact on male fertility.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154261"},"PeriodicalIF":4.6,"publicationDate":"2025-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144827229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Importance of post-translational protein modifications in PFAS toxicity","authors":"Narukkottil Safreena ,&nbsp;Goutam Chandra ,&nbsp;Venkatesh P. Thirumalaikumar ,&nbsp;Jason Cannon","doi":"10.1016/j.tox.2025.154260","DOIUrl":"10.1016/j.tox.2025.154260","url":null,"abstract":"<div><div>Per- and polyfluoroalkyl substances (PFAS) are a group of synthetic compounds known for their high chemical stability, resistance to degradation. This class includes thousands of different compounds widely used in various products like non-stick cookware, water-repellent fabrics, food packaging, firefighting foam, and many personal care products. While these properties contribute to their industrial utility, they also pose significant environmental and health concerns due in part to long environmental and biological half-lives. PFAS contamination has been widely reported in water sources and soil, along with reports of bioaccumulation in the blood and tissues of living organisms. These chemicals have been linked to a range of health effects, including potential risks to cancer, neurotoxicity, and developmental disorders. Given their broad involvement in disease pathology, it is critical to investigate the underlying molecular mechanisms of PFAS toxicity. This review evaluates current research on the impact of various PFAS on different proteins, their post-translational modifications (PTMs), and the effects on cellular signalling pathways. It also reviews proteomic studies conducted over the years that aim to elucidate these protein-level changes, while highlighting existing gaps in the field. Despite growing attention to PFAS toxicity, significant advances are required in elucidating the primary mechanisms of toxic action. Studying PFAS effects from a proteome perspective could be very promising. In particular, the lack of organ-specific proteomic data, including studies on distinct brain regions and PTM-specific profiling, represents a critical area for future research.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154260"},"PeriodicalIF":4.6,"publicationDate":"2025-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144810536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Maternal, fetal, and neonatal toxicity and potency estimates of perfluorohexane sulfonate (PFHxS) from oral maternal exposure in the Sprague-Dawley rat","authors":"Justin M. Conley ,&nbsp;Christy S. Lambright ,&nbsp;Nicola Evans ,&nbsp;Jacqueline Bangma ,&nbsp;Jermaine Ford ,&nbsp;Donna Hill ,&nbsp;Elizabeth Medlock Kakaley ,&nbsp;L. Earl Gray Jr.","doi":"10.1016/j.tox.2025.154245","DOIUrl":"10.1016/j.tox.2025.154245","url":null,"abstract":"<div><div>Perfluorohexane sulfonate (PFHxS) is a legacy, long-chain per- and polyfluoroalkyl substance (PFAS) frequently detected in human serum and environmental media across the globe. Previously published developmental studies in rats indicated a general lack of effects in common endpoints like maternal and offspring body and liver weights at doses up to 50 mg/kg/d. To facilitate mixture-based studies of PFAS co-exposure, individual dose response parameters are necessary for model predictions. To address these data needs we exposed timed-pregnant Sprague-Dawley rat dams to PFHxS (3–125 mg/kg/d) via oral gavage from gestation days (GD)14–18 or GD8 to postnatal day (PND)2. We evaluated a range of key events and adverse outcomes in both exposure intervals with a focus on chemical effects on serum thyroid hormone concentrations and liver toxicity. Maternal exposure from GD14–18 resulted in increased maternal liver weight (≥30 mg/kg), reduced serum total thyroxine (T4; ≥10 mg/kg) and low-density lipoprotein (LDL; ≥10 m/kg), and modest changes in maternal liver fatty acid metabolism-associated and fetal liver peroxisome proliferator-activated receptor signaling-associated gene expression. Exposure from GD8-PND2 did not affect newborn liver glycogen concentration, but did result in other neonatal effects consistent with prior PFAS studies including reduced pup survival (125 mg/kg), reduced pup bodyweight (≥62.5 mg/kg), increased pup liver weight (≥10 mg/kg), highly elevated pup serum total cholesterol and bile acids (≥3 mg/kg) and reduced serum thyroid hormones (T3, rT3, T4; ≥3 mg/kg). Maternal effects were largely limited to reductions in serum total T3 and T4 (≥10 mg/kg) and elevated serum BUN (≥62.5 mg/kg). Compared to previously published data for other PFAS, PFHxS displayed relatively low potency as a function of maternal serum concentration across a range of effects.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154245"},"PeriodicalIF":4.6,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144776236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A high-throughput microphysiological system to quantify key events leading to liver fibrosis","authors":"Saskia Schmidt ,&nbsp;Laura Suter-Dick","doi":"10.1016/j.tox.2025.154248","DOIUrl":"10.1016/j.tox.2025.154248","url":null,"abstract":"<div><div>New approach methodologies (NAMs), including microphysiological systems (MPS), are emerging as alternatives to animal testing. In the liver, chronic hepatocellular damage can progress to fibrosis, which has been described by an Adverse Outcome Pathway (AOP). However, standardized in vitro models that capture and quantify key AOP events and cell-cell interactions are lacking. We developed a scalable liver fibrosis model using the 384-well Akura™ Twin microplate featuring 168 interconnected well pairs. We studied fibrosis progression by seeding HepaRG microtissues (MTs), with or without THP-1 cells in alpha wells and hepatic stellate cell (hTERT-HSC) MTs in beta wells. Cell health and metabolic activity were monitored via specific sensors that detect glucose and lactate levels. Transforming growth factor beta 1 (TGF-β1), methotrexate (MTX) and acetaminophen (APAP) reduced albumin production, indicating hepatocellular injury. TGF-β1 activated THP-1, increasing ALOX5AP, TREM2, and TGF-β1 mRNA expression. PAI-1 protein levels increased following treatment with TGF-β1, particularly in HepaRG-THP-1 co-cultures. In hTERT-HSCs, TGF-β1 also induced expression of fibrosis markers (ACTA2, COL1A1, COL3A1 and FN1) and increased stress fibers and fibronectin expression. Extracellular matrix remodeling was confirmed by elevated Pro-Collagen 1A1 and CTGF protein levels upon TGF-β1 treatment. The Akura™ Twin platform enables high-throughput modeling of liver fibrosis, mimicking the key events of the liver fibrosis AOP. This model, combining a high-throughput MPS with established cell lines, offers a promising tool to investigate fibrosis mechanisms and advancing quantitative AOP development. Journal: Toxicology (Special Issue: Hepatotoxicity: mechanisms and animal-free prediction models).</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154248"},"PeriodicalIF":4.6,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144776235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhalation of dust accumulated on personal computer fans in the office may cause pathological effects by disrupting the metabolic activity of alveolar macrophages","authors":"Wonkyun Jung ,&nbsp;Cheolho Yoon ,&nbsp;Mi-Jin Yang ,&nbsp;Taekyung Yu ,&nbsp;Hyosun Choi ,&nbsp;Nam Chul Kwon ,&nbsp;Seong-Jin Choi ,&nbsp;Beom-Geon Kim ,&nbsp;Sung-Min Hong ,&nbsp;Cheon Woong Choi ,&nbsp;Ji Ae Lee ,&nbsp;Eun-Jung Park","doi":"10.1016/j.tox.2025.154247","DOIUrl":"10.1016/j.tox.2025.154247","url":null,"abstract":"<div><div>Considering that computer fans draw in dust from the office air, and that printers and copiers are typically placed near computers, we collected dust that accumulated on computer fans from three commercial personal computer centers to predict the health effects of inhaled office dust (OD). OD contained various types of microplastics and non-protein particles, and several hydrophilic elements were bound to the OD. When aspirated OD via the pharynx for 90 days, blood levels of IP and K significantly decreased in male mice, but not in female mice. Meanwhile, the total number of pulmonary cells increased, accompanied by increases in the proportions of lymphocytes and PMNs, as well as pulmonary levels of pro-inflammatory and anti-inflammatory mediators. Foamy alveolar macrophages and mucous cell hyperplasia were observed in the lung tissues, accompanied by increased levels of TGF-β1, IL-11, and IL-24. More importantly, structural disruption of mitochondria, mitophagy, and the formation of lamellar bodies and collagen fibers were frequently observed in the lung and heart tissues of OD-treated mice. In addition, LDH release and ATP production were not significantly affected in OD-treated alveolar macrophage cells, even at the highest concentration. Meanwhile, very interestingly, the activity of the NADPH-dependent cellular oxidoreductase enzyme decreased dramatically, even at the lowest concentration. Genes related to cytokine-cytokine receptor interaction and neutrophil extracellular trap formation were most significantly affected in cells exposed to OD, and Saa3, Cybb, Slc7a11, and CXCL2 were identified as the most upregulated genes. The expression of SOD-2, FeH, and COX-2 proteins was enhanced in OD-treated cells, along with increased NO production, whereas the expression of transferrin receptor and BAX protein was reduced in the same cells. Collectively, we suggest that OD exposure may increase the lung burden of foreign bodies by disrupting the metabolic activity of alveolar macrophages, ultimately resulting in chronic pulmonary diseases. We also hypothesize that impaired self-renewal capacity may contribute to the lung burden of OD.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154247"},"PeriodicalIF":4.6,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144769122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mathematical modelling reveals compound-specific stress pathway activity","authors":"Elsje J. Burgers ,&nbsp;Tamara Y. Danilyuk ,&nbsp;Raju P. Sharma ,&nbsp;Nadine Renner ,&nbsp;Andreas Verlohner ,&nbsp;Nicole Rocker ,&nbsp;Philipp Ternes ,&nbsp;Lukas S. Wijaya ,&nbsp;Marcel Leist ,&nbsp;Peter Bouwman ,&nbsp;Franziska M. Zickgraf ,&nbsp;Stefan Schildknecht ,&nbsp;Bob van de Water ,&nbsp;Joost B. Beltman","doi":"10.1016/j.tox.2025.154234","DOIUrl":"10.1016/j.tox.2025.154234","url":null,"abstract":"<div><div>Drug-induced liver injury (DILI) is a major problem for the drug development industry. It has been suggested that activation of stress pathways within cells is an important indicator of DILI. In this project, we aimed to develop a mathematical model of invoked stress responses by three compounds with high DILI liability: nitrofurantoin, diclofenac and ketoconazole. To this end, we used imaging data from HepG2 cells and cell-associated compound and intracellular glutathione measurements. We initially developed a model for the integrated and oxidative stress responses following nitrofurantoin exposure. Subsequently, we expanded this to simulate responses to diclofenac and ketoconazole. To apply the model to these compounds multiple parameters required recalibration, yet the structure of the model was unchanged. Our analysis shows that the magnitude of interactions between transcription factors and downstream targets can differ even when the activated pathways are the same.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"518 ","pages":"Article 154234"},"PeriodicalIF":4.6,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144765576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ethanol exposure impaired mitotic division in apical radial glial cells and disrupted early cortical development in human forebrain organoids: Implications for ethanol-induced microcephaly","authors":"Lanhai Lü ,&nbsp;Miao Bai ,&nbsp;Xiaoqing Wang ,&nbsp;Jie Liu ,&nbsp;Xiaoyang Wu ,&nbsp;Huang-ge Zhang ,&nbsp;Wenke Feng ,&nbsp;Shao-yu Chen","doi":"10.1016/j.tox.2025.154244","DOIUrl":"10.1016/j.tox.2025.154244","url":null,"abstract":"<div><div>Maternal alcohol exposure can severely disrupt the differentiation of neural stem/progenitor cells, leading to a reduced number of neurons and decreased volume and thickness of the cortical layers, a hallmark feature commonly associated with microcephaly. Recent studies from our laboratory have demonstrated that ethanol exposure interfered with the formation of radial glial processes and impaired the generation and migration of outer radial glial cells in human forebrain organoids. However, the precise mechanisms by which ethanol affects the production of outer radial glial cells remain unclear. In this study, human forebrain organoids were used to investigate the effects of ethanol exposure on the mitotic divisions of radial glial cells and cortical development. Our findings revealed that ethanol exposure significantly reduced mitotic cell division in apical radial glial cells. Exposure to ethanol also significantly decreased symmetric cell division while increasing asymmetrical cell division in these cells. Additionally, ethanol exposure reduced the number of outer radial glial cells in forebrain organoids, suggesting that ethanol exposure impaired the production of outer radial glial cells by disrupting the mitotic division of apical radial glial cells. Our findings also revealed that ethanol-induced disruptions in mitotic cell division in apical radial cells and the subsequent reduction in outer radial glial cells contributed to the reduced thickness of the developing cortical wall and resulted in smaller forebrain organoids. This study significantly advances our understanding of the cellular mechanisms underlying ethanol-induced microcephaly and provides potential avenues for developing novel interventions to prevent or mitigate the neurodevelopmental deficits associated with prenatal alcohol exposure, including ethanol-induced microcephaly.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"517 ","pages":"Article 154244"},"PeriodicalIF":4.6,"publicationDate":"2025-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144722219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Computerized predictive approaches of genotoxicity and mutagenesis in 3D Hepoid of normal and transformed human hepatocytes","authors":"Sophie Rose ,&nbsp;Frédéric Ezan ,&nbsp;Ludovic Huot ,&nbsp;Thierry Pécot ,&nbsp;Medjda Bellamri ,&nbsp;Robert Turesky ,&nbsp;Fabrice Nesslany ,&nbsp;Anne Platel ,&nbsp;Sophie Langouët","doi":"10.1016/j.tox.2025.154242","DOIUrl":"10.1016/j.tox.2025.154242","url":null,"abstract":"<div><div>The low specificity of standard <em>in vitro</em> genotoxicity tests in mammalian cells continues to necessitate animal experimentation, raising ethical concerns that conflict with the principles of the 3Rs (Replacement, Reduction, and Refinement). This underscores the urgent need for reliable alternative <em>in vitro</em> assays. Given the liver's critical role in xenobiotic detoxification and bioactivation, we developed an advanced 3D <em>in vitro</em> model of human hepatocytes that supports the proliferation and long-term differentiation of primary human hepatocytes and HepaRG cells. Using a range of methodologies, including the γH2AX assay, the comet assay, the micronucleus test, and transcriptomic analysis, we investigated DNA damage induced by well-known genotoxic carcinogens such as methylmethane sulfonate (MMS), mitomycin C (MMC), colchicine, vinblastine, 1,2-dimethylhydrazine (DMH), and aflatoxin B₁ (AFB₁), as well as non-genotoxic carcinogens, including di-2-ethylhexyl phthalate (DEHP) and methylcarbamate. Our data demonstrate the reliability of the 3D human hepatocyte Hepoid model in replicating <em>in vivo</em> results. We effectively discriminated between genotoxic and non-genotoxic carcinogenic molecules using complementary methods analyzed with advanced computational approaches. We further studied the <em>in vitro</em> genotoxic potential of four main Heterocyclic Aromatic Amines (HAAs) including 2-amino-9<em>H</em>-pyrido[2,3-<em>b</em>]indole (AαC), 2-amino-3-methylimidazo[4,5-<em>f</em>]quinoline (IQ), 2- amino-3,8-dimethylimidazo[4,5-<em>f</em>]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-<em>b</em>]pyridine (PhIP), which are food-borne and environmental contaminants suspected to contribute to hepatocellular carcinoma. Our results validate the Hepoid model as a reliable <em>in vitro</em> system for assessing the genotoxic and mutagenic risks of chemicals and confirm the ability of HAAs to induce DNA damage in a highly differentiated human liver model.</div></div>","PeriodicalId":23159,"journal":{"name":"Toxicology","volume":"517 ","pages":"Article 154242"},"PeriodicalIF":4.6,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144718729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}