Toxicology in Vitro最新文献

{"title":"Cigarette sidestream smoke-induced cellular senescence and the protective role of histone H2AX","authors":"Yukako Komaki,&nbsp;Yuko Ibuki","doi":"10.1016/j.tiv.2025.106076","DOIUrl":"10.1016/j.tiv.2025.106076","url":null,"abstract":"<div><div>Cigarette smoke imposes serious health hazards such as cancer and cardiovascular diseases but is also associated with cellular senescence. Recently, histone loss and modifications are reported as one of the characteristics of cellular senescence. In this study, we examined the relationship between cigarette smoke-induced cellular senescence and histone H2A variant H2AX, an important player in DNA damage response. We exposed normal human skin diploid fibroblast ASF-4-1 to cigarette sidestream smoke (CSS) extract and successfully induced premature senescence. Persistent DNA damages are known to induce cellular senescence. Double strand breaks (DSBs) formation was detected in CSS-treated cells, indicating DSBs could be the cause for the CSS-induced cellular senescence. In the senescent cells, persistent phosphorylation of histone H2AX (γ-H2AX) and unexpected increase of H2AX protein expression was observed. To elucidate the role of H2AX in CSS-induced cellular senescence, we depleted H2AX in ASF-4-1 cells with siRNA. In H2AX-depleted cells, CSS-induced elevated β-galactosidase activity was more prominent. CSS concentration-dependent increase of reactive oxygen species and DSBs formation was also facilitated by H2AX depletion. These results suggest that histone H2AX may have a protective role against DNA damage-induced premature senescence.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106076"},"PeriodicalIF":2.6,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143891621","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":"Association between drug-induced heart failure and CYP1A1, CYP1B1, and CYP3A4 inhibition: Utility of cytochrome P450 inhibition assay for evaluating cardiotoxicity of drug candidates","authors":"Shunnosuke Kaito ,&nbsp;Kiyomi Sato ,&nbsp;Takamitsu Sasaki ,&nbsp;Takuomi Hosaka ,&nbsp;Ryota Shizu ,&nbsp;Jun-ichi Takeshita ,&nbsp;Kouichi Yoshinari","doi":"10.1016/j.tiv.2025.106075","DOIUrl":"10.1016/j.tiv.2025.106075","url":null,"abstract":"<div><div>Unpredictable adverse drug reactions (ADRs) present a significant challenge in drug development and require advanced prediction systems for ADRs. Previously, we identified a connection between drug-induced liver injury (DILI) and the inhibition of CYP1A1 or CYP1B1, reporting the usefulness of this inhibition data from these cytochrome P450s (P450s) for predicting DILI. This study aimed to investigate the utility of P450 inhibition data in predicting drug-induced organ toxicities beyond DILI. We selected 364 drugs with ADR information as test drugs from the public database SIDER (Side Effect Resource). Our focus was on 10 groups of ADRs affecting the liver, kidney, heart, blood/hematopoietic system, intestines, muscle, and lungs, as classified by MedDRA. The inhibitory activities of 10 human P450s were evaluated in vitro using recombinant enzymes and luminescent substrates. Our analyses revealed notable associations between heart failure and the inhibition of CYP1A1, CYP1B1, and CYP3A4. Heart failure-positive drugs tended to exhibit strong inhibition of these P450s compared to heart failure-negative drugs. Furthermore, most drugs that inhibited two or three of the three P450 forms were found to be heart failure-positive. These results suggest that the inhibition assay data for CYP1A1, CYP1B1, and CYP3A4 help assess drug-induced cardiotoxicity during drug development.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106075"},"PeriodicalIF":2.6,"publicationDate":"2025-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869489","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":"A549 cells exposed to a marijuana smoke extract affect mono-layer integrity related to oxidative stress state","authors":"Elvira Gómez-Guerrero ,&nbsp;Yazmín Debray-García ,&nbsp;Octavio Gamaliel Aztatzi-Aguilar ,&nbsp;Fanny Azuzena Colchero-Amateco ,&nbsp;Omar Amador-Muñoz ,&nbsp;Josefina Poblano-Bata ,&nbsp;Irais Poblete-Naredo ,&nbsp;Arnulfo Albores","doi":"10.1016/j.tiv.2025.106072","DOIUrl":"10.1016/j.tiv.2025.106072","url":null,"abstract":"<div><div>Marijuana smoke contains several toxic compounds that may induce dysregulation of oxidative mechanisms and barrier system in airway alveolar cells. This study aimed to assess whether marijuana smoke extract (MSE) modifies mono-layer integrity and antioxidant effects on the alveolar epithelial cells. A549 cells were exposed to MSE (0.1 to 5 μg/mL) or cannabinoid (+)-WIN 55,212–2 (WIN; 0.01 to 1 μM) for 24 h. Epithelial integrity and protein expression of claudin (Cl)-2, Cl-5, and occludin (Ocl) were evaluated by transepithelial electrical resistance (TEER), permeability assay, Western blot, immunofluorescence, and qPCR. TEER decreased after MSE or WIN exposure, whereas the monolayer permeability increased. Protein expression and localization of Cl-2 and Ocl were reduced after MSE treatment. However, WIN increased Cl-2 protein and decreased Cl-5 and Ocl. Differential gene expressions were observed between treatments. Malondialdehyde (MDA) production and advanced oxidation protein products (AOPP) determination showed that MSE increased AOPP, whereas WIN augmented the MDA production and decreased AOPP levels. The activity of antioxidant enzymes shows an increase in catalase, glutathione-<em>S</em>-transferase, γ-glutamyl transferase and arginase after MSE treatment and a decrease with WIN. Data indicates that MSE exposure alters epithelial integrity and the alveolar cells antioxidant response that, finally, may lead to lung damage.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106072"},"PeriodicalIF":2.6,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143874686","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":"Effects of Penthorum chinense pursh and gallic acid on embryonic and cardiac development in zebrafish (Danio rerio)","authors":"Kehui Liu ,&nbsp;Hui Jiang ,&nbsp;Rong Ji ,&nbsp;Yuanyuan Ma ,&nbsp;Rui Zhang ,&nbsp;Binbin Song ,&nbsp;Ying Han","doi":"10.1016/j.tiv.2025.106074","DOIUrl":"10.1016/j.tiv.2025.106074","url":null,"abstract":"<div><div><em>Penthorum chinense</em> Pursh (<em>P. chinense</em>), known for its anti-inflammatory and antioxidant properties, is valued for its low toxicity in animal and human models. However, concerns have arisen regarding the developmental effects of its bioactive components. This study investigates the acute toxicity of <em>P. chinense</em> extract and gallic acid on zebrafish embryos. The calculated LC₅₀ values were 237.0 mg/L for <em>P. chinense</em> extract and 328.4 mg/L for gallic acid, demonstrating a dose-response relationship with increasing mortality rates. Developmental assessments revealed significant morphological abnormalities, including heart defects, swim bladder and tail malformations, particularly at higher concentrations. Body length and eye diameter showed a hormetic dose-response to <em>P. chinense</em> extract, with increased growth at lower concentrations but a decrease at higher doses. Cardiac evaluations revealed altered heart rates, initially increasing and then decreasing at elevated concentrations. qRT-PCR analyses confirmed modulation of several heart-related genes, highlighting the differential impacts on cardiac development. These findings underscore the need to carefully assess the potential risks of <em>P. chinense</em> extract and gallic acid exposure in aquatic organisms.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106074"},"PeriodicalIF":2.6,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143864611","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":"Species differences in microsomal metabolism of hydroxychloroquine","authors":"Guilin Wei ,&nbsp;Chaohua Yan ,&nbsp;Liwei Zou ,&nbsp;Yong Liu ,&nbsp;Ling Yang","doi":"10.1016/j.tiv.2025.106063","DOIUrl":"10.1016/j.tiv.2025.106063","url":null,"abstract":"<div><div>Hydroxychloroquine (HCQ), a medication renowned for its anti-malarial and anti-autoimmune properties, is susceptible to drug-drug interactions (DDIs), particularly those involving cytochrome P450 enzymes (CYPs). The pharmacokinetic and metabolic profiles of HCQ across species are not well-characterized. In this study, we conducted a comparative analysis of HCQ metabolism and pharmacokinetics in human liver microsomes and those from various preclinical animal models. Metabolite profiling indicated that the mono-oxidized metabolite M5 was predominantly produced in microsomes from mouse (MLM), rats (RLM), dogs (DLM), and pigs (PLM). Conversely, two other oxidative metabolites, M4 and M6, were uniquely generated in PLM. Utilizing selective inhibitors of human CYP isoforms, we identified key enzymes in HLM that differ from those in other species. Furthermore, the metabolic pathways in HLM were distinct from those observed in other species. In HLM, two metabolic pathways have been identified, each comprising a two-step reaction. CYP2D6, CYP2C8, CYP3A4, and CYP1A1 are the key enzymes involved in HCQ metabolism, with HCQ demonstrating significant substrate inhibition of CYP2D6, CYP2C8, and CYP1A1. Our work shed a new light on selection of suitable experimental animal models for accurate evaluation of HCQ's <em>in vivo</em> processes and its potential in multi-drug regimens.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106063"},"PeriodicalIF":2.6,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143844908","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":"Corrigendum to “Tetrazolium–based colorimetric assays underestimat the direct antitumor effects of bevacizumab” [Toxicology in vitro 91 (2023) 105631]","authors":"Pei Wei ,&nbsp;Min Wang ,&nbsp;Mao Lin ,&nbsp;Zhiyong Wang","doi":"10.1016/j.tiv.2025.106061","DOIUrl":"10.1016/j.tiv.2025.106061","url":null,"abstract":"","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106061"},"PeriodicalIF":2.6,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042928","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":"Antibiotics reduce intestinal bile acid reuptake in an in vitro model system","authors":"Nina Zhang,&nbsp;Véronique M.P. de Bruijn,&nbsp;Weijia Zheng,&nbsp;Wouter Bakker,&nbsp;Bennard van Ravenzwaay,&nbsp;Ivonne M.C.M. Rietjens","doi":"10.1016/j.tiv.2025.106071","DOIUrl":"10.1016/j.tiv.2025.106071","url":null,"abstract":"<div><div>Enterohepatic circulation of bile acids is a highly efficient process that is important for bile acid homeostasis. The aim of the present study was to characterize the impact of a series of antibiotics (lincomycin, streptomycin, vancomycin and tobramycin) on the intestinal reuptake of conjugated bile acids (TCA, TCDCA, GCA and GCDCA) using a Caco-2 <em>in vitro</em> transwell model system. The results obtained demonstrate that both pre-exposure and co-exposure of the cells to an antibiotic and the bile acids, affected bile acid transport, to an extent that depended on the antibiotic, its concentration and the type of conjugated bile acid tested. Tobramycin, at concentrations in line with dose levels at which this antibiotic induced effects on bile acid homeostasis <em>in vivo</em>, appeared able to inhibit bile acid transport after pre-exposure of the cells, likely resulting from an effect on the expression of bile acid transporters <em>via</em> its effects on protein synthesis at ribosome level. Upon co-exposure of the Caco-2 cells to an antibiotic and the bile acids, all four antibiotics appeared to significantly reduce the transport of especially the conjugated bile acids TCDCA and GCDCA with a potency that decreased in the order vancomycin &gt; tobramycin = streptomycin &gt; lincomycin. The effects observed illustrate the possibility of using a new approach methodology (NAM) to study effects on intestinal bile acid reuptake.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106071"},"PeriodicalIF":2.6,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143844907","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":"Integrative toxicity assessment of tocotrienol-rich fraction from palm oil using in silico methods and zebrafish embryotoxicity model","authors":"Thenmoly Damodaran ,&nbsp;Najib Sani Yahaya ,&nbsp;Mohd Nizam Mordi","doi":"10.1016/j.tiv.2025.106062","DOIUrl":"10.1016/j.tiv.2025.106062","url":null,"abstract":"<div><div>Tocotrienol-rich fraction (TRF), a natural form of vitamin E derived from palm oil, possesses antioxidant properties. However, its potential embryonic developmental toxicity remains unclear. This study investigated TRF's toxicity using <em>in silico</em> methods and zebrafish embryos. Zebrafish embryos were exposed to TRF (31.25 to 2000 μg/mL) for 96 h post-fertilization (hpf). Mortality, hatching rate, heart rate, and morphological malformations were assessed at 24, 48, 72, and 96 hpf. <em>In silico</em> analysis predicted good pharmacokinetic properties and minimal side effects for five TRF constituents, except for hERG II inhibition, which is associated with cardiac toxicity. TRF exposure up to 96 hpf showed no embryotoxicity in zebrafish at ≤1000 μg/mL. However, TRF at concentrations of ≥1000 μg/mL significantly inhibited hatching rate at 72 hpf, indicating a delay in the hatching process. Additionally, 1000 μg/mL of TRF resulted in reduced heart rate and hypopigmentation in the embryos. Moreover, higher TRF concentrations (≥500 μg/mL) caused morphological malformations including spinal curvature, pericardial edema, and yolk sac edema, in the embryos. These findings suggest that TRF from palm oil is likely safe at concentrations below 500 μg/mL during embryonic development. However, the potential effects of long-term exposure and chronic toxicity warrant further investigation to ensure safety during early pregnancy.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106062"},"PeriodicalIF":2.6,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143792093","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":"Ozone and particulate matter co-exposure at the air-liquid interface: Establishing an approach to assess pollutant interactions in vitro","authors":"Andres R. Henriquez ,&nbsp;Marjolaine Godbout-Cheliak ,&nbsp;Alain Filiatreault ,&nbsp;Errol M. Thomson","doi":"10.1016/j.tiv.2025.106060","DOIUrl":"10.1016/j.tiv.2025.106060","url":null,"abstract":"<div><h3>Background</h3><div>Air pollution is a complex mixture of gases and particulates that varies spatially and temporally, making attribution of adverse effects to specific individual air pollutants a challenge. To disentangle effects of mixtures in a controlled setting, reproducible and realistic co-exposures of human-relevant models to gaseous and particulate pollutants are needed. Although air-liquid interface (ALI) exposures offer considerable promise as non-animal models for inhalation toxicity testing, a lack of studies comparing individual and co-exposures to gaseous and particulate pollutants has thus far prevented assessment of their strengths and limitations for disentangling effects of pollutant mixtures.</div></div><div><h3>Methods</h3><div>Using an integrated ALI exposure system, we characterized the interaction between ozone and particles (25 nm fluorescent polystyrene beads) to assess effects on and reproducibility of critical physical endpoints including temperature, humidity, ozone concentration and particle deposition. Particle deposition and concentration were assessed <em>via</em> three independent methods: fluorescence, quartz crystal microbalance (QCM), and airborne particle count. To evaluate the acute biological effects of an air pollutant mixture <em>in vitro</em>, human lung type 2 epithelial-like cells (A549) were exposed at the ALI to air, ozone (O₃), particles, and O₃ + particles (co-exposure) for 1 h (<em>n</em> = 4 independent repeats/exposure type). Cell injury and inflammation were quantified by extracellular lactate dehydrogenase (LDH) activity and release of proinflammatory cytokines (interleukin (IL)-8 and IL-6) respectively 0 and 24 h post-exposure.</div></div><div><h3>Results</h3><div>Exposures were effective at delivering targeted O₃ exposures under controlled temperature and relative humidity. In-well particle deposition and airborne concentration exiting the exposure system, quantified through parallel methods, were consistent, and increased in relation to aerosolized particle concentration. Levels of each pollutant were effectively maintained in the presence of the other. O₃ alone, and co-exposure to O₃ and particles, increased LDH release from A549 cells, indicating pollutant-specific cytotoxicity. In contrast, IL-8 and IL-6 release (24 h &gt; 0 h) were not changed by exposure to the individual pollutants, but tended to increase following co-exposure.</div></div><div><h3>Conclusion</h3><div>The present work establishes the utility of ALI exposure systems to disentangle individual effects of pollutants from a mixture, and highlights the importance of direct experimental characterization of dosimetry and exposure conditions.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106060"},"PeriodicalIF":2.6,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143782013","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":"The effect of organic solvents on the in vitro transport activity of three OATP isoforms","authors":"Rina Saito ,&nbsp;Takeshi Akiyoshi ,&nbsp;Kazunari Tsujii ,&nbsp;Riho Takahashi ,&nbsp;Hiroki Kataoka ,&nbsp;Ayuko Imaoka ,&nbsp;Hisakazu Ohtani","doi":"10.1016/j.tiv.2025.106059","DOIUrl":"10.1016/j.tiv.2025.106059","url":null,"abstract":"<div><h3>Purpose</h3><div><em>In vitro</em> studies of transporter activity often require the addition of organic solvents such as dimethyl sulfoxide (DMSO), methanol, and ethanol, to dissolve substrates and/or inhibitors. Although this may attenuate the transport activity, the influence of different types of organic solvents on transporter activity has not been elucidated. This study aimed to quantitatively assess the impact of organic solvents on the transport activity of organic anion transporting polypeptide (OATP) 1B1, 1A2, and 2B1.</div></div><div><h3>Methods</h3><div>The concentration-dependent influence of DMSO, methanol, and ethanol on the uptake of [³H]estrone 3-sulfate mediated by OATP1A2, OATP2B1 or 2′,7′-dichlorofluorescein (DCF) mediated by OATP1B1 was assessed using HEK293 cells expressed the respective OATP protein and the concentration that reduced the uptake by 20 % (IC₂₀) was calculated.</div></div><div><h3>Results and discussion</h3><div>The uptake activities of OATPs were reduced by methanol and ethanol in a concentration-dependent manner, with IC₂₀ values of 2.4–3.4 % and 0.51–3.8 % respectively. In contrast, DMSO did not affect the OATP2B1-mediated uptake at the maximum concentration (10 %) of DMSO whereas it exhibited concentration-dependent inhibition for OATP1B1 and 1A2 with IC₂₀ values of approximately 3 % and 0.7 %, respectively. This study provides useful information regarding experimental conditions for evaluating OATPs activity <em>in vitro</em>.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106059"},"PeriodicalIF":2.6,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143755942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}