Toxicology in Vitro最新文献_第7页

Comparison of the behavior of human lung epithelial cell lines cultured at the air-liquid interface and assessment of their responses after benzo(a)pyrene exposure 在气液界面培养的人肺上皮细胞系的行为比较及其暴露于苯并(a)芘后的反应评估

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-23 DOI: 10.1016/j.tiv.2025.106122

Suen Boulé, Louise Verhaeghe, Dominique Courcot, Yann Landkocz

The biological effects of air pollution are still not well known, due to its complex mixture of particulate and gaseous compounds. In vitro cell culture models exposed at the air-liquid interface (ALI) represent a potential alternative to in vivo experiments to assess the effects of outdoor air pollution. This study compares two bronchial cell lines, Calu-3 and BEAS-2B, and two alveolar cell lines, hAELVi and A549, regarding their capacity to form a tight epithelial cell barrier for a 2-week culture period and metabolize xenobiotics actively. Culture at the air-liquid interface permits the Calu-3 and hAELVi cells to form and maintain a tight epithelial cell barrier with lower permeability to lucifer yellow, greater trans-epithelial electrical resistance, and the presence of Zonula Occludens 1 (ZO-1) protein at the membrane, than the BEAS-2B and A549 cells. Exposure to benzo(a)pyrene (BaP) induces the up-regulation of CYP1A1 and CYP1B1 genes, proteins, and functional activity at the air-liquid interface in all cell lines. So, these results demonstrate that the Calu-3 and the hAELVi cells are the more relevant models to assess the effects of ambient air pollution at the air-liquid interface, forming a tight epithelial cell barrier and being metabolically active.

空气污染的生物效应仍然不为人所知，因为它是微粒和气体化合物的复杂混合物。暴露于空气-液体界面（ALI）的体外细胞培养模型代表了评估室外空气污染影响的体内实验的潜在替代方案。本研究比较了两种支气管细胞系Calu-3和BEAS-2B以及两种肺泡细胞系hAELVi和A549在2周培养期内形成紧密上皮细胞屏障和积极代谢外源药物的能力。与BEAS-2B和A549细胞相比，在气液界面培养使Calu-3和hAELVi细胞形成并维持紧密的上皮细胞屏障，其对路西法黄的渗透性较低，跨上皮电阻较大，膜上存在ZO-1蛋白。暴露于苯并(a)芘（BaP）可诱导所有细胞系中CYP1A1和CYP1B1基因、蛋白和气液界面功能活性的上调。因此，这些结果表明，Calu-3和hAELVi细胞在气液界面形成紧密的上皮细胞屏障并具有代谢活性，是评估环境空气污染影响的更合适的模型。

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引用次数: 0

Polystyrene nanoplastic co-exposed to BPA and BPS induces cytotoxicity, genotoxicity, and alters ROS production in HepG2 cells 聚苯乙烯纳米塑料共同暴露于BPA和BPS诱导细胞毒性，遗传毒性，并改变HepG2细胞的ROS产生

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-23 DOI: 10.1016/j.tiv.2025.106121

Cecilia Cristina de Souza Rocha , Caroline Andolfato Sanchez , Marília Cristina Oliveira Souza , Estéfani Maria Treviso , Gabriel Henrique Savietto , Paula Pícoli Devóz , Lucas Victor Pereira de Freitas , Lusânia Maria Greggi Antunes , Jonas Augusto Rizzato Paschoal , Fernando Barbosa Junior

During plastic degradation, it is fragmented into micro and nanoplastic, which can adsorb contaminants from the environment, increasing the plastics´ toxicity. Bisphenol A is used in plastic production and can be an endocrine disruptor. Bisphenol S is an analog of bisphenol A and has been used as an alternative to “BPA-free” products. Therefore, this is the first research proposing to verify whether nanoplastics associated with bisphenol A or S can increase toxicity in HepG2 cells. Nanoplastics associated with bisphenols could alter the cell viability of HepG2 cells compared to the group treated with the nanoplastic alone and concerning the respective bisphenol. The co-exposure of nanoplastic to bisphenols A or S promoted cytotoxic and genotoxic damage in HepG2 cells, altering the reactive oxygen species production, increasing DNA strand breaks, and increasing apoptotic cells. Bisphenols A and S also showed cytotoxic and genotoxic effects in HepG2 cells. However, 8-OHdG was only detected in the group treated with the nanoplastic at the lowest concentration. This study highlights the cytotoxicity and genotoxicity of nanoplastics and bisphenols A and S, providing new insights into hepatocyte toxicity.

在塑料降解过程中，它被分解成微塑料和纳米塑料，它们可以吸附环境中的污染物，增加塑料的毒性。双酚A用于塑料生产，可能是一种内分泌干扰物。双酚S是双酚A的类似物，已被用作“无双酚A”产品的替代品。因此，这是首次提出验证纳米塑料与双酚A或S相关是否会增加HepG2细胞毒性的研究。与单独处理纳米塑料和双酚相关的组相比，与双酚相关的纳米塑料可以改变HepG2细胞的活力。纳米塑料与双酚A或S的共暴露促进了HepG2细胞的细胞毒性和基因毒性损伤，改变了活性氧的产生，增加了DNA链断裂，增加了凋亡细胞。双酚A和S对HepG2细胞也有细胞毒性和基因毒性作用。然而，8-OHdG仅在最低浓度的纳米塑料处理组中检测到。本研究强调了纳米塑料和双酚A和S的细胞毒性和遗传毒性，为肝细胞毒性提供了新的见解。

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引用次数: 0

The neuroprotective role of chlorogenic acid and Fisetin in differentiated neuronal cell line-SHSY5Y against amyloid-β-induced neurotoxicity 绿原酸和非西汀对分化神经细胞系shsy5y抗淀粉样蛋白β诱导的神经毒性的保护作用。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-20 DOI: 10.1016/j.tiv.2025.106110

Apoorv Sharma , Puneet Kumar , Asimul Islam , Monika Bhardwaj , Vijay Kumar , Hridayesh Prakash

Alzheimer's disease (AD) is a neurodegenerative disorder and characterized by amyloid-beta (Aβ) accumulation, synaptic dysfunction, oxidative stress, and lacks effective therapies. Fisetin and chlorogenic acid (CGA) are natural polyphenols have shown potential in mitigating several age-related diseases. Therefore this study investigates their neuroprotective effects against Aβ1–42-induced toxicity in differentiated SHSY5Y cells. Both Fisetin and CGA reversed the deleterious effects of Aβ1–42 by restoring redox balance, suppressing reactive oxygen species, and upregulating mRNA expression of critical antioxidant enzymes like SOD1, GSR, and CAT. These compounds also attenuated Aβ1–42-induced mitophagy via reduced PINK1 expression and restored mitochondrial fusion by upregulating MFN2. Autophagy-related pathways were significantly modulated which was evidenced by increased PRKAA1 (AMPK) and decreased MTOR mRNA levels, alongside elevated expression of ATG101, ATG13, ULK1, SQSTM1 (p62) and reduced ATG5 levels. Fisetin and CGA improved synaptic integrity by upregulating DLG4 (PSD95) and SYP (synaptophysin) and reducing ACHE (acetylcholinesterase) expression. These findings highlight their potential in ameliorating Aβ1–42-induced neuronal toxicity through autophagy activation, synaptic preservation, and mitochondrial function enhancement. Furthermore, our docking studies also revealed good Fisetin and CGA binding affinity within AMPK and mTOR's binding pocket (FKBP12-FRB). Although the neuroprotective effects of CGA and Fisetin are underscored by transcriptional and docking studies, further translational and biophysical validation is required to demonstrate their therapeutic efficacy against AD-related neurodegeneration.

阿尔茨海默病（AD）是一种神经退行性疾病，以β淀粉样蛋白（a β）积累、突触功能障碍、氧化应激为特征，缺乏有效的治疗方法。非瑟酮和绿原酸（CGA）是天然的多酚类物质，在缓解几种与年龄有关的疾病方面显示出潜力。因此，本研究探讨了它们对a - β1-42诱导的SHSY5Y分化细胞的神经保护作用。非瑟酮和CGA均通过恢复氧化还原平衡、抑制活性氧以及上调SOD1、GSR和CAT等关键抗氧化酶mRNA表达来逆转a - β1-42的有害作用。这些化合物还通过降低PINK1表达来减弱a β1-42诱导的线粒体自噬，并通过上调MFN2来恢复线粒体融合。PRKAA1 （AMPK）升高，MTOR mRNA水平降低，ATG101、ATG13、ULK1、SQSTM1 （p62）表达升高，ATG5水平降低，这些都证明了自噬相关通路的显著调节。非西汀和CGA通过上调DLG4 （PSD95）和SYP （synaptophysin）以及降低ACHE（乙酰胆碱酯酶）表达来改善突触完整性。这些发现强调了它们通过自噬激活、突触保存和线粒体功能增强来改善a β1-42诱导的神经元毒性的潜力。此外，我们的对接研究还显示，在AMPK和mTOR的结合口袋（FKBP12-FRB）中，fissetin和CGA具有良好的结合亲和力。尽管转录和对接研究强调了CGA和非塞汀的神经保护作用，但还需要进一步的翻译和生物物理验证来证明它们对ad相关神经变性的治疗效果。

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引用次数: 0

Beyond traditional toxicology: The transformative power of PBTK modeling 超越传统毒理学：PBTK建模的变革力量

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-18 DOI: 10.1016/j.tiv.2025.106111

Anagha Damre, Aniruddha Banerjee

Physiologically Based Toxicokinetic (PBTK) modeling has emerged as a crucial tool in toxicokinetics, enabling the quantitative assessment of chemical absorption, distribution, metabolism, and excretion (ADME) across various biological systems. Unlike traditional toxicokinetic approaches, PBTK models integrate physiological and biochemical parameters to allow for precise interspecies and dose extrapolations. This capability enhances their applicability in regulatory risk assessment for pharmaceuticals, industrial chemicals, food additives, cosmetics, and pesticides. High Throughput Toxicokinetics (HTTK) and in vitro-to-in vivo extrapolations (IVIVE) further improve the predictive power of PBTK models by utilizing large-scale experimental datasets and computational approaches. Additionally, these models facilitate route-to-route extrapolations, predicting systemic exposure across different administration routes such as oral, inhalation, and dermal pathways. PBTK modeling also enables the estimation of specific target tissue concentrations, cross-species extrapolations, and extrapolations to special populations, thereby improving human biological modeling. Furthermore, the integration of PBTK models in ecological risk assessment supports the evaluation of environmental chemical exposure effects on diverse species. As regulatory agencies increasingly adopt PBTK models for toxicity evaluations, their role in advancing data-driven risk assessment and reducing reliance on animal testing continues to grow. This review explores the application of PBTK modeling in toxicokinetics and its alignment with regulatory guidelines for risk assessment and interspecies extrapolation.

基于生理的毒物动力学（PBTK）建模已成为毒物动力学的重要工具，可以定量评估各种生物系统中的化学吸收、分布、代谢和排泄（ADME）。与传统的毒物动力学方法不同，PBTK模型整合了生理和生化参数，以允许精确的种间和剂量外推。这种能力增强了它们在药品、工业化学品、食品添加剂、化妆品和农药监管风险评估中的适用性。高通量毒物动力学（HTTK）和体外到体内外推（IVIVE）通过利用大规模实验数据集和计算方法进一步提高了PBTK模型的预测能力。此外，这些模型促进了途径到途径的外推，预测了不同给药途径（如口服、吸入和皮肤途径）的全身暴露。PBTK建模还可以估计特定的目标组织浓度，跨物种外推，以及对特殊种群的外推，从而改进人类生物学建模。此外，PBTK模型在生态风险评价中的整合，为评价环境化学物质暴露对不同物种的影响提供了支持。随着监管机构越来越多地采用PBTK模型进行毒性评估，它们在推进数据驱动的风险评估和减少对动物试验的依赖方面的作用不断增强。这篇综述探讨了PBTK模型在毒性动力学中的应用，以及它与风险评估和种间外推的监管指南的一致性。

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引用次数: 0

Inter-laboratory validation study of an in vitro glucocorticoid receptor transactivation assay for testing potential endocrine disrupting chemicals 用于检测潜在内分泌干扰物的体外糖皮质激素受体反激活试验的实验室间验证研究。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-17 DOI: 10.1016/j.tiv.2025.106109

Andrea Rivero-Arze , Marina Grimaldi , Géraldine Maillet , Cindie Gesbert , Erwan Michelin , Clémentine Garoche , Emmanuelle Maillot-Maréchal , Olivier Palluel , François Sermier , Sebastian Hoffmann , Jérôme Couteau , Philippe Hubert , Elise Grignard , Selim Aït-Aïssa , Patrick Balaguer , Torben Österlund

The glucocorticoid receptor (GR) belongs to the family of steroid receptors (SRs). These receptors regulate a vast selection of cell-, tissue-, and organism biology, and are also targets of endocrine disrupting chemicals warranting design and validation of in vitro assays. Here we report a “blinded” ring trial of an in vitro cell-based GR transactivation assay with four involved laboratories. The laboratories set up the assay and tested 34 selected chemicals with remarkably good concordance. There was agreement between all laboratories for the classification of activity in 97 % of the cases, and three or more laboratories were always in agreement. The within laboratory concordance was very high (99.6 %) with only one of all 272 triplicates deviating. The assay was, thus, deemed easily transferable and reproducible within and between laboratories, since they would arrive at the same qualitative results. Furthermore, for the chemicals with solid data regarding GR activation or inhibition, the laboratories arrived at the expected conclusion in all cases. Overall, the transfer and validation were successful, and the method is under evaluation to become an OECD test guideline. The method is expected to become valuable in tiered approaches for assessing chemicals or environmental samples together with other similar methods.

糖皮质激素受体（GR）属于类固醇受体（SRs）家族。这些受体调节着大量的细胞、组织和有机体生物学，也是内分泌干扰化学物质的目标，需要设计和验证体外试验。在这里，我们报告了一个“盲法”环试验的体外细胞为基础的GR转激活测定与四个相关实验室。实验室建立了测定法，并对34种选定的化学物质进行了测试，一致性非常好。在97% %的病例中，所有实验室对活动性的分类是一致的，三个或更多的实验室总是一致的。实验室内一致性非常高（99.6 %），272个三重复中只有一个偏离。因此，该分析被认为易于在实验室内部和实验室之间转移和重复，因为它们会得到相同的定性结果。此外，对于具有关于GR激活或抑制的可靠数据的化学品，实验室在所有情况下都得出了预期的结论。总的来说，转移和验证是成功的，该方法正在评估中，以成为经合组织的测试指南。该方法有望与其他类似方法一起在评估化学品或环境样品的分层方法中变得有价值。

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引用次数: 0

Lupeol-loaded PLGA particles conserve anti-topoisomerase activity and decrease the cytotoxicity of lupeol 负载lupeol的PLGA颗粒保持抗拓扑异构酶活性，降低lupeol的细胞毒性。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-15 DOI: 10.1016/j.tiv.2025.106107

Francisco Fabián Razura-Carmona , Mayra Herrera-Martínez , Jorge Alberto Sánchez-Burgos , Alejandro Pérez-Larios , Karina Janice Guadalupe Díaz-Reséndiz , Manuel Iván Girón-Pérez , Marco Vinicio Ramírez-Mares

Lupeol (LP), a bioactive triterpene, has been extensively studied for its chemoprotective, anti-inflammatory, and anti-arthritic properties. To enhance its bioavailability, lupeol-loaded polylactic-co-glycolic acid (PLGA) nanoparticles, designated T_LP14, were developed using the emulsification-evaporation method. These nanoparticles exhibited an average size of 339.8 nm, a polydispersity index (PdI) of 0.240, and an encapsulation efficiency of 38 %. In topoisomerase II inhibition assays, T_LP14 retained activity comparable to that of lupeol free, indicating that encapsulation does not impair its biological function. In vitro cytotoxicity assays on BEAS-2 and HEPG2 cell lines demonstrated that concentrations above 1250 μg/mL of T_LP14 induced minimal cytotoxic effects, suggesting low toxicity in non-tumor cells. Furthermore, ex vivo studies on peripheral blood mononuclear cells showed that a 1000 μg/mL concentration of free lupeol induced 28.01 % apoptosis, while the encapsulated formulation significantly reduced this adverse effect. These findings support the potential of T_LP14 nanoparticles as an effective controlled-release system for lupeol, enhancing its safety profile while preserving its therapeutic activity.

Lupeol （LP）是一种生物活性三萜，因其化学保护、抗炎和抗关节炎的特性而被广泛研究。为了提高其生物利用度，采用乳化蒸发法制备了载聚乳酸-羟基乙酸（PLGA）纳米粒子TLP14。这些纳米颗粒的平均尺寸为339.8 nm，多分散指数（PdI）为0.240，包封效率为38 %。在拓扑异构酶II抑制实验中，TLP14保留了与不含lupeol的活性相当的活性，这表明包封不会损害其生物学功能。对BEAS-2和HEPG2细胞株的体外细胞毒实验表明，浓度高于1250 μg/mL的TLP14对非肿瘤细胞的细胞毒作用最小，表明其毒性较低。此外，对外周血单个核细胞的体外研究表明，1000 μg/mL浓度的游离lup柚醇可诱导28.01 %的细胞凋亡，而包封制剂可显著降低这种不良反应。这些发现支持了TLP14纳米颗粒作为一种有效的lupeol控释系统的潜力，增强了其安全性，同时保持了其治疗活性。

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引用次数: 0

Testing chemicals for CYP19 inhibition: Comparison of a modified H295R steroidogenesis assay to a commercial fluorometric enzyme inhibition kit 检测CYP19抑制的化学物质：改良的H295R甾体生成试验与商业荧光酶抑制试剂盒的比较。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-13 DOI: 10.1016/j.tiv.2025.106108

Caroline Despicht, Kieu-Mi Tran, Terje Svingen, Anna Kjerstine Rosenmai

Aromatase (CYP19A1) is a key enzyme that converts androgens to estrogens. Given its central role in estrogen biosynthesis, aromatase is a target of endocrine disruption. Aromatase disruption can be measured in vitro with cell-free enzyme assays or using cells expressing CYP19A1. Results may vary between models, as cell-based models can have adaptive responses not present in enzyme assays. Here, we have compared 21 chemicals for their ability to affect aromatase by direct and indirect disruption using an aromatase commercial kit and a modified H295R steroidogenesis assay. New data was generated for eighteen test compounds in either one or both of the models. Chemicals eliciting a response could be divided into five categories: 1) substances that decreased the response in both models, 2) substances that directly inhibited aromatase, but increased 17β-estradiol (E2) levels in H295R assay, 3) substances that directly inhibited aromatase, but did not affect E2 levels in H295R assay, 4) substances that did not inhibit aromatase activity, but increased E2 levels in the H295R model, and 5) substances that did not alter responses in either of the assays. These results illustrate that both assays are useful for assessing the potential of chemicals to affect E2 levels either through direct aromatase inhibition or other indirect mechanisms.

芳香化酶（CYP19A1）是将雄激素转化为雌激素的关键酶。鉴于其在雌激素生物合成中的核心作用，芳香化酶是内分泌干扰的目标。芳香酶破坏可以通过体外无细胞酶测定或使用表达CYP19A1的细胞来测量。模型之间的结果可能有所不同，因为基于细胞的模型可能具有酶分析中不存在的适应性反应。在这里，我们比较了21种化学物质通过直接和间接破坏芳香化酶的能力，使用芳香化酶商业试剂盒和改进的H295R甾体生成试验。在一种或两种模型中产生了18种测试化合物的新数据。引起反应的化学物质可分为五类：1)在两种模型中均降低反应的物质；2)直接抑制芳香化酶，但在H295R实验中增加17β-雌二醇（E2）水平的物质；3)直接抑制芳香化酶，但在H295R实验中不影响E2水平的物质；4)不抑制芳香化酶活性，但在H295R模型中增加E2水平的物质；5)在两种实验中均不改变反应的物质。这些结果表明，这两种分析方法都可用于评估化学物质通过直接芳香化酶抑制或其他间接机制影响E2水平的潜力。

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引用次数: 0

Impact of gadolinium oxide based nanoparticles on genotoxic, cytotoxic and oxidative stress in human lymphocytes 氧化钆纳米颗粒对人淋巴细胞基因毒性、细胞毒性和氧化应激的影响。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-13 DOI: 10.1016/j.tiv.2025.106106

Isela Álvarez-González , Lizbeth Espinosa-García , Felipe de Jesús Carrillo-Romo , Antonieta García-Murillo , José A. Morales-González , Eduardo O. Madrigal-Santillán , Eduardo Madrigal-Bujaidar

Gadolinium-based nanoparticles are presently applied in biomedicine and industry, and their potential is increasing. In this report we synthetized three nanoparticles by the sol-gel, supercritical drying method: Gd₂O₃, Gd₂O₃ doped with Eu⁺³, and Gd₂O₃ doped with Eu⁺³and functionalized with thenoyltrifluoroacetone (TTA). The nanoparticles were characterized for luminescence, morphology, size, and Z potential. The results showed that the highest luminescence was reached with the TTA functionalized nanoparticle, all of them had a porous net structure that decreased to 21 nm in the last nanoparticle, and the Z potential was found from −2 to −12 mV. To determine their geno/cytotoxic potential we performed the cytochalasine-block micronucleus cytome, and the MTT assays in human lymphocytes. With these assays, we demonstrated that the two main genotoxic effects were the nuclear buds and nucleoplasmic bridges, and that nanoparticles decreased the cellular proliferation, mainly because of the induction of apoptosis (about 70 %) in contrast with the 30 % of necrosis. Finally, a significant lipid and protein oxidation increase was determined in the nanoparticles. Our results suggest that the observed geno/cytotoxic damage may be related with the presence of oxidative stress.

目前，钆基纳米颗粒在生物医学和工业上的应用越来越广泛，其应用潜力越来越大。本文采用溶胶-凝胶、超临界干燥的方法合成了三种纳米粒子：Gd2O3、掺杂Eu+3的Gd2O3和掺杂Eu+3并经乙烯基三氟丙酮（TTA）功能化的Gd2O3。对纳米粒子的发光、形貌、尺寸和Z电位进行了表征。结果表明，TTA功能化纳米粒子的发光强度最高，均呈多孔网状结构，最后一个纳米粒子的发光强度降至21 nm， Z电位在-2 ~ -12 mV之间。为了确定它们的基因/细胞毒性潜能，我们在人淋巴细胞中进行了细胞查拉辛阻断微核细胞组和MTT测定。通过这些实验，我们证明了两种主要的基因毒性作用是核芽和核质桥，纳米颗粒减少了细胞增殖，主要是因为诱导了细胞凋亡（约70% %），而不是30% %的坏死。最后，纳米颗粒中脂质和蛋白质氧化显著增加。我们的研究结果表明，观察到的基因/细胞毒性损伤可能与氧化应激的存在有关。

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引用次数: 0

E171-induced toxicity in human iPSC-derived colon organoids: Effects on cell viability, ROS generation, DNA damage, and gene expression changes e171诱导的人类ipsc衍生的结肠类器官毒性：对细胞活力、ROS生成、DNA损伤和基因表达变化的影响

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-10 DOI: 10.1016/j.tiv.2025.106105

Nicolaj S. Bischoff , Anna K. Undas , Marcel van Herwijnen , Marcha Verheijen , Jacco J. Briedé , Simone G. van Breda , Dick T.H.M. Sijm , Theo M. de Kok

Food-grade titanium dioxide (E171) is a widely used food additive with debated safety, particularly regarding its genotoxic effects. This study assessed the dose-dependent toxicity of E171 in human induced pluripotent stem cell (iPSC)-derived colon organoids. Organoids were exposed to E171 (0.1–1000 μg/mL) for 24 h, and effects on cell viability, reactive oxygen species (ROS) generation, DNA damage, and gene expression were evaluated.

Results showed no impact on cell viability but a dose-dependent increase in ROS formation, peaking at 1000 μg/mL. Electrospin Resonance Spectroscopy (ESR) showed a dose-dependent increase in ROS, with increased E171 concentrations. The alkaline comet assay revealed significant DNA damage from 100 μg/mL, with oxidative DNA damage detected at 10 μg/mL using formamidopyrimidine DNA glycosylase (FPG). RNA sequencing identified differentially expressed genes (DEGs) at 100 and 250 μg/mL, linked to translational activity, signal transduction, and DNA damage repair. Gene set enrichment analysis (GSEA) indicated activation of ribosome, chemical carcinogenesis–ROS, and metabolic pathways (carbon metabolism, glycolysis), while key regulatory pathways (Wnt, MAPK, PI3K-Akt) were suppressed.

These findings suggest that E171 induces oxidative stress and DNA damage, modulating transcriptomic pathways associated with metabolism, proliferation, and cancer. Further research is necessary to determine its long-term effects on human gastrointestinal health.

食品级二氧化钛（E171）是一种广泛使用的食品添加剂，安全性存在争议，特别是关于其基因毒性的影响。本研究评估了E171对人诱导多能干细胞（iPSC）衍生的结肠类器官的剂量依赖性毒性。将E171 （0.1 ~ 1000 μg/mL）暴露于类器官24 h，观察其对细胞活力、活性氧（ROS）生成、DNA损伤和基因表达的影响。结果显示，对细胞活力没有影响，但ROS形成呈剂量依赖性增加，在1000 μg/mL时达到峰值。电自旋共振光谱（ESR）显示，随着E171浓度的增加，ROS呈剂量依赖性增加。碱性彗星实验显示，当浓度为100 μg/mL时，DNA损伤显著；当浓度为10 μg/mL时，甲脒嘧啶DNA糖基化酶（FPG）检测到DNA氧化损伤。RNA测序鉴定出100和250 μg/mL浓度的差异表达基因（DEGs），这些差异表达基因与翻译活性、信号转导和DNA损伤修复有关。基因集富集分析（GSEA）显示核糖体、化学致癌- ros和代谢途径（碳代谢、糖酵解）被激活，而关键调控途径（Wnt、MAPK、PI3K-Akt）被抑制。这些发现表明，E171诱导氧化应激和DNA损伤，调节与代谢、增殖和癌症相关的转录组通路。需要进一步的研究来确定其对人体胃肠道健康的长期影响。

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引用次数: 0

The protective mechanisms of glycyrrhizic acid for deoxynivalenol-induced toxicity in rabbit corneal stroma 甘草酸对脱氧雪腐镰刀醇致兔角膜基质毒性的保护机制。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-04 DOI: 10.1016/j.tiv.2025.106104

Chunlan Liang , Xuewei Xiong , Qi Shi , Qingqing Li , Guocheng Yu , Jingxiang Zhong , Lian Liu

Objectives

This study aimed to investigate the effects and mechanisms of deoxynivalenol (DON) on rabbit corneal stroma cell culture and observe the protective effect of glycyrrhizic acid (GA) against DON-induced damage.

Methods

Rabbit corneal stromal cells (RCSCs) were isolated and divided into groups: control group (CON), 0.5 mmol/L GA (GA group), 400 ng/mL DON (DON group), and 0.5 mmol/L GA + 400 ng/mL DON (GAD). The effect of GA on RCSCs induced by DON was evaluated using various assays, including CCK-8 assay, Flow cytometry, qPCR, and Western blot.

Results

DON inhibited RCSCs proliferation, increased apoptosis, and raised the proportion of cells in the S and G2/M phase. It raised reactive oxygen species (ROS) levels and reduced mitochondrial membrane potential (MMP). Additionally, DON upregulated Bax, Caspase-3, inflammatory factors (IL-1α, IL-1β, TNF-α), and matrix metalloproteinases (MMP-1/8), while downregulating tissue inhibitors of matrix metalloproteinases (TIMP-1) and type I collagen. In contrast, GA partially restored RCSCs proliferation and reduced apoptosis, inflammation, and collagen degradation.

Conclusions

DON can induce toxicity in RCSCs, promoting the expression of inflammatory factors, disrupting the balance between MMP-1/8 and TIMP-1, and promoting type I collagen degradation. Conversely, GA reduces the oxidative stress damage, cell apoptosis, and inflammatory reaction of RCSCs induced by DON, and reduces the degradation of type I collagen.

目的：研究脱氧雪腐镰刀菌醇（DON）对兔角膜基质细胞培养的影响及其机制，并观察甘草酸（GA）对DON损伤的保护作用。方法：分离兔角膜基质细胞（RCSCs），分为对照组（CON）、0.5 mmol/L GA组（GA组）、400 ng/mL DON组（DON组）、0.5 mmol/L GA + 400 ng/mL DON （GAD）。采用CCK-8、流式细胞术、qPCR、Western blot等多种检测方法评价GA对DON诱导的RCSCs的影响。结果：DON抑制RCSCs增殖，增加凋亡，提高S期和G2/M期细胞比例。升高活性氧（ROS）水平，降低线粒体膜电位（MMP）。此外，DON上调Bax、Caspase-3、炎症因子（IL-1α、IL-1β、TNF-α）和基质金属蛋白酶（MMP-1/8），下调基质金属蛋白酶（TIMP-1）和I型胶原的组织抑制剂。相反，GA可以部分恢复RCSCs的增殖，减少细胞凋亡、炎症和胶原降解。结论：DON可诱导RCSCs毒性，促进炎症因子的表达，破坏MMP-1/8与TIMP-1之间的平衡，促进I型胶原降解。相反，GA可减轻DON诱导的RCSCs氧化应激损伤、细胞凋亡和炎症反应，减少I型胶原的降解。

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