Toxicology in Vitro最新文献_第2页

Effects of cyanobacterial metabolites and their mixtures on biomarkers of oxidative stress in RTgill-W1 cells 蓝藻代谢产物及其混合物对RTgill-W1细胞氧化应激生物标志物的影响

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2026-01-06 DOI: 10.1016/j.tiv.2026.106195

Adam Bownik, Barbara Pawlik-Skowrońska

The aim of our investigation was to determine the effects of seven cyanobacterial metabolites microcystin-LR (MC-LR), anabaenopeptin-A (ANA-A), cylindrospermopsin (CYL), anabaenopeptin-B (ANA-B) aeruginosin 98 A (AER-A), aeruginosin 98B (AER-B), microginin-FR1 (MG-FR1) and their mixtures on common indicators of oxidative stress in RTgill-W1cells. The cells were exposed to the metabolites at various concentrations and the following parameters were determined after 48 h: catalase (CAT) activity, lipid peroxidation (LPO), total nitric oxide (NO) level and superoxide (SOD) dismutase activity. Data were analyzed with the use of one-way ANOVA (N = 3) and Dunnett's test. We found that all single tested metabolites increased but CYL decreased CAT activity. Mixtures had stimulatory effect, however antagonistic interactions were found in the binary and ternary mixtures. No lipid peroxidation occurred in the cells exposed to any of the tested variant. Only AER-A increased NO level, however the rest of both single metabolites at highest concentrations (1004 nM) and mixtures reduced the level of this parameter. Only ANA-B inhibited SOD activity at the highest concentration, however no alterations were found in the cells exposed to binary or ternary mixtures. The study showed that cyanobacterial metabolites may induce oxidative stress in fish gill cells, however effects are dependent on type of a metabolite and concentration of a component in mixtures. On the basis of antagonistic interactions in ternary mixture it may be hypothesized that during natural exposure components of mixtures may reciprocally mitigate their effects.

本研究旨在探讨7种蓝藻代谢产物微囊藻毒素- lr （MC-LR）、鸭绿霉素-A （ANA-A）、柱状精子素（CYL）、鸭绿霉素- b （ANA-B）、绿脓杆菌蛋白98 A （AER-A）、绿脓杆菌蛋白98B （AER-B）、微球蛋白- fr1 （MG-FR1）及其混合物对rtgill - w1细胞氧化应激指标的影响。将细胞暴露于不同浓度的代谢物中，48 h后测定过氧化氢酶（CAT）活性、脂质过氧化（LPO）、总一氧化氮（NO）水平和超氧化物歧化酶（SOD）活性。数据分析采用单因素方差分析（N = 3）和Dunnett检验。我们发现所有单一测试的代谢物都增加了，但CYL降低了CAT活性。混合物具有刺激作用，但在二元和三元混合物中发现拮抗相互作用。暴露于任何测试变体的细胞中均未发生脂质过氧化。只有AER-A增加了NO水平，而最高浓度（1004 nM）和混合浓度下的其他单代谢物均降低了该参数的水平。只有ANA-B在最高浓度下抑制SOD活性，但在二元或三元混合物中未发现任何变化。研究表明，蓝藻代谢物可能诱导鱼鳃细胞氧化应激，但影响取决于代谢物的类型和混合物中成分的浓度。根据三元混合物中的拮抗相互作用，可以假设在自然暴露期间，混合物的组分可以相互减轻其作用。

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引用次数: 0

Dual effects of metformin and resveratrol on compromising viability of endometrial cancer cells 二甲双胍和白藜芦醇对子宫内膜癌细胞生存能力的双重影响

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2026-01-06 DOI: 10.1016/j.tiv.2026.106194

Henrique Leal de Oliveira , Sara Hartke , Victória Borgmann A. de Souza , Carolina Vaccari Batista , Gabriela Pasqualim , Vânia Marisia Santos Fortes dos Reis , Edison Capp , Leo Anderson Meira Martins , Ilma Simoni Brum

Raising of mitogenic and anti-apoptotic agents – such as insulin, insulin-like growth factor type 1, and estrogen – during obesity and diabetes mellitus (types 1 and 2) favors the endometrial cancer (EC) development. Metformin, commonly used for treating type 2 diabetes, and resveratrol, a natural polyphenol, can both decrease cancer cell proliferation by modulating the PI3K/Akt/mTOR pathway. We evaluate the effects of metformin and/or resveratrol in an in vitro model of human type 1 endometrioid EC. Ishikawa cells were treated with 0.1 to 50 mM of metformin and/or 0.1 to 75 μM of resveratrol from 24 h to 72 h. Analyses assessed cell viability, cytotoxicity, caspases activation, mitochondrial function, cellular death, cell cycle, and the PI3K/Akt/mTOR pathway gene expression. In-silico analysis was conducted using Cytoscape. Metformin induced mitochondrial swelling, caspase-mediated apoptosis, and cell cycle arrest. Resveratrol decreased mitochondrial mass, cytotoxicity, and induced cell cycle arrest. Combined treatment with the highest concentrations reduced mitochondrial activity, cytotoxicity, and caspase activation while maintaining apoptotic features and cell cycle arrest. Resveratrol attenuated the toxic effects of metformin but it could be inducing a caspase-independent cell death in co-treated cells. Although in-silico analysis suggested potential molecular targets and interconnected mechanisms, lower concentrations did not alter PI3K/Akt/mTOR gene expression.

在肥胖和糖尿病（1型和2型）期间，有丝分裂和抗凋亡因子如胰岛素、胰岛素样生长因子1型和雌激素的升高有利于子宫内膜癌（EC）的发展。通常用于治疗2型糖尿病的二甲双胍和天然多酚白藜芦醇都可以通过调节PI3K/Akt/mTOR通路来抑制癌细胞增殖。我们评估二甲双胍和/或白藜芦醇对人类1型子宫内膜样EC体外模型的影响。石川细胞用0.1至50 mM的二甲双胍和/或0.1至75 μM的白藜芦醇处理24至72小时。分析评估细胞活力、细胞毒性、半胱天冬酶激活、线粒体功能、细胞死亡、细胞周期和PI3K/Akt/mTOR通路基因表达。使用Cytoscape进行了硅内分析。二甲双胍诱导线粒体肿胀、caspase介导的细胞凋亡和细胞周期阻滞。白藜芦醇减少线粒体质量、细胞毒性和诱导细胞周期阻滞。最高浓度的联合治疗降低了线粒体活性、细胞毒性和半胱天冬酶活性，同时维持了凋亡特征和细胞周期停滞。白藜芦醇减轻了二甲双胍的毒性作用，但它可能在共处理的细胞中诱导caspase非依赖性细胞死亡。尽管计算机分析提示了潜在的分子靶点和相互关联的机制，但较低的浓度并未改变PI3K/Akt/mTOR基因的表达。

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引用次数: 0

Diesel exhaust nanoparticles: Cellular adaptation in lung epithelial and fibroblast cells – An in vitro study 柴油废气纳米颗粒：肺上皮细胞和成纤维细胞的细胞适应性-体外研究。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2026-01-03 DOI: 10.1016/j.tiv.2026.106193

Gabriel Martínez-Razo , Ruth Angélica Lezama , Armando Vega-López , María Lilia Domínguez-López

Diesel exhaust particles (DEnP) represent a major urban air pollutant with known adverse effects on human health, yet detailed cellular interactions at the nanoscale are poorly understood. This study aims to elucidate the cellular responses and adaptation mechanisms of human lung epithelial and fibroblast-like cell lines exposed to Mexican diesel exhaust nanoparticles. Exhaust samples from cold and warm engine emissions were analyzed for polyaromatic hydrocarbons, organic matter, and elemental composition using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Additionally, the DEnP nanostructure was scrutinized using 3D topographical image analysis. In vitro assays assessed cell proliferation, adhesion molecule expression (ICAM-1, VCAM-1), and proteins related to endocytosis (clathrin and dynamin) in response to DEnP exposure. SEM and TEM analyses revealed distinct nanoparticle forms and compositions, with significant increases in cell proliferation, endocytosis, and adhesion molecule expression observed, suggesting robust cellular adaptation mechanisms to counteract DEnP-induced stress. The study confirms significant cellular adaptations in response to DEnP, underscoring the need for preventive strategies to mitigate the impacts of exposure. These findings provide a foundation for further investigation into long-term cellular adaptations and their implications for pulmonary health.

柴油废气颗粒（DEnP）是一种主要的城市空气污染物，已知对人类健康有不利影响，但在纳米尺度上详细的细胞相互作用知之甚少。本研究旨在阐明墨西哥柴油机尾气纳米颗粒对人肺上皮细胞和成纤维细胞样细胞系的细胞反应和适应机制。利用扫描电子显微镜（SEM）和透射电子显微镜（TEM）对冷热发动机排放的废气样品进行了多芳烃、有机物和元素组成的分析。此外，使用三维地形图像分析仔细检查了DEnP纳米结构。体外实验评估了DEnP暴露后的细胞增殖、粘附分子表达（ICAM-1、VCAM-1）和与内吞作用相关的蛋白质（网格蛋白和动力蛋白）。扫描电镜和透射电镜分析揭示了不同的纳米颗粒形式和组成，观察到细胞增殖、内吞作用和粘附分子表达显著增加，表明细胞适应机制可以抵抗denp诱导的应激。该研究证实了对DEnP的显著细胞适应，强调了采取预防策略以减轻暴露影响的必要性。这些发现为进一步研究长期细胞适应及其对肺部健康的影响提供了基础。

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引用次数: 0

Barrier gel formulations and coated gloves to reduce skin permeation of nicotine and protect against green tobacco sickness 屏障胶配方和涂层手套，以减少皮肤对尼古丁的渗透，防止绿色烟草病。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-22 DOI: 10.1016/j.tiv.2025.106192

Abhay U. Andar , Youcheng Liu , Dana C. Hammell , David A. Sterling , Tom Klingner , Mark Tokarski , Mark Boeniger , Audra Stinchcomb

Tobacco harvesting workers may have high levels of skin exposure to nicotine that can lead to green tobacco sickness. Current exposure reduction methods are often infeasible. The purpose of this work was to develop and evaluate the effectiveness of topical barrier gel formulations as a personal protective equipment to reduce nicotine permeation through skin. Four formulations of a barrier gel developed and applied on Yucatan miniature pig skin were tested using a PermeGear flow through in vitro diffusion apparatus. Donor solutions of either L-nicotine or green tobacco leaf extract with and without the use of barrier gel formulations were analyzed over a 24 h exposure period. High pressure liquid chromatography was used to quantify the nicotine content in the receiver compartment. Gloves coated with a barrier gel formulation were also tested. The best barrier gel formulations reduced in vitro skin permeation of nicotine by 97.6 % from L-nicotine, by 64.0 % from green tobacco leaf extract, and by 86.6 % from green tobacco leaf extract for gardening gloves coated with the barrier gel. The barrier gel is effective in reducing skin permeation of nicotine in vitro and might have greater preventive capabilities at environmental exposure levels of nicotine during tobacco harvesting.

烟草采收工人的皮肤可能高度暴露于尼古丁，从而导致绿烟病。目前减少暴露的方法往往是不可行的。这项工作的目的是开发和评估局部屏障凝胶制剂作为个人防护设备的有效性，以减少尼古丁通过皮肤的渗透。开发并应用于尤卡坦微型猪皮肤的四种屏障凝胶配方，使用pergear体外扩散装置进行了测试。在24 h的暴露时间内，对l -尼古丁或绿色烟草叶提取物的供体溶液进行了分析，其中有或没有使用屏障凝胶配方。采用高压液相色谱法定量烟碱含量。涂有屏障凝胶配方的手套也进行了测试。在涂有屏障凝胶的园艺手套中，最佳屏障凝胶配方可使l -尼古丁的体外皮肤渗透率降低97.6% %，使绿烟叶提取物的体外皮肤渗透率降低64.0 %，使绿烟叶提取物的体外皮肤渗透率降低86.6% %。屏障凝胶在体外有效地减少尼古丁的皮肤渗透，并且在烟草收获期间的尼古丁环境暴露水平下可能具有更大的预防能力。

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引用次数: 0

Interactions of neocryptotanshinone and human cytochrome P450 in silico and in vitro 新隐丹参酮与人细胞色素P450的相互作用。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-17 DOI: 10.1016/j.tiv.2025.106191

Xiu Chen , Xiaoyi Pan , Jieping Zhao , Huihui Jin , Hengbin Zhang , Yongbiao Song , Hui Zhou , Jianbiao Yao , Huidi Jiang

Neocryptotanshinone (NCTS), an ingredient of Salviae Miltiorrhizae Radix et Rhizoma, is a promising compound for development since it exhibits various pharmacological effects including hypoglycemic and anti-inflammatory activities. In this study, we aimed to elucidate the interactions between NCTS and cytochrome P450s (CYPs), including the CYP-mediated NCTS metabolism and NCTS-induced CYP regulation. The results revealed that NCTS was mainly metabolized by CYPs in human liver microsomes (HLMs), and CYP2C8 and 2C9 were identified as the main CYPs involved; the relative contributions of CYP2C8 and 2C9 were 35 % and 65 %, respectively, after normalization of individual CYP abundance in the human liver. Meanwhile, NCTS weakly inhibited CYP1A2, 2C8, and 2C9, with IC₅₀ > 30 μM. In addition, NCTS induced CYP2B6 and CYP3A4 expression in human primary hepatocytes, and the underlying mechanism was found by the activation of the pregnane X receptor (PXR) vis reporter gene assay. Molecular docking of NCTS and CYPs, PXR confirmed these results. Our study sheds light on the interactions of NCTS with CYPs and provides useful information for predicting potential drug-drug interactions for NCTS, which will be helpful for NCTS development and clinical utilization of drugs containing NCTS.

新crypto丹参酮（NCTS）是丹参的一种成分，具有降血糖、抗炎等多种药理作用，是一种很有开发前景的化合物。在本研究中，我们旨在阐明NCTS与细胞色素p450 （CYPs）之间的相互作用，包括CYPs介导的NCTS代谢和NCTS诱导的CYP调控。结果表明，NCTS主要被人肝微粒体（HLMs）中的CYPs代谢，其中CYP2C8和2C9是主要参与代谢的CYPs；在人肝脏个体CYP丰度归一化后，CYP2C8和2C9的相对贡献分别为35 %和65 %。NCTS对CYP1A2、2C8、2C9的抑制作用较弱，IC50 > 30 μM。此外，NCTS在人原代肝细胞中诱导CYP2B6和CYP3A4的表达，并通过激活妊娠X受体（PXR）报告基因实验发现其潜在机制。NCTS与CYPs的分子对接，PXR证实了这些结果。本研究揭示了NCTS与CYPs的相互作用，为预测NCTS的潜在药物相互作用提供了有用的信息，这将有助于NCTS的开发和含NCTS药物的临床应用。

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引用次数: 0

Activation of Toll-like receptor 2 reveals microbial contamination beyond endotoxins on micro- and nanoplastics toll样受体2的激活揭示了微和纳米塑料上的内毒素以外的微生物污染。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-13 DOI: 10.1016/j.tiv.2025.106190

Øyvind P. Haugen , Itziar Polanco-Garriz , Victor Alcolea-Rodriguez , Raquel Portela , Rita Bæra , Hamed Sadeghiankaffash , Jana Hildebrandt , Dmitri Ciornii , Korinna Altmann , Francesco Barbero , Ivana Fenoglio , Julián J. Reinosa , José F. Fernández , Alberto Katsumiti , Laura M.A. Camassa , Håkan Wallin , Shan Zienolddiny-Narui , Anani K. Afanou

Current literature on health hazards associated with micro- and nanoplastics (MNPs) is largely influenced by studies that insufficiently account for potential microbial contamination of their test materials. This may lead to misinterpretation of outcomes, as the test materials may be incorrectly considered pristine MNPs. The present study screened eight MNP test materials for microbial contaminants using Toll-like receptor (TLR) reporter cells for TLR2 and TLR4 and the commonly used Limulus amebocyte lysate (LAL) assay. Our results show that MNPs testing negative for endotoxins, based on the absence of TLR4 activation and negative LAL results, may still contain microbial ligands that selectively activate TLR2. Moreover, five of the eight MNP test materials contained microbial ligands capable of activating TLR2 and/or TLR4. Compared to the LAL assay, TLR4-based screening effectively detected endotoxin contamination. Overall, we found that the TLR reporter cell assay provides broader coverage than the LAL assay in detecting microbial ligands, which appear to be highly prevalent in MNP test materials.

目前关于与微和纳米塑料（MNPs）相关的健康危害的文献在很大程度上受到研究的影响，这些研究没有充分考虑其测试材料的潜在微生物污染。这可能导致对结果的误解，因为测试材料可能被错误地认为是原始MNPs。本研究利用toll样受体（TLR）报告细胞对TLR2和TLR4以及常用的鲎试剂（LAL）法筛选了8种MNP微生物污染物检测材料。我们的研究结果表明，内毒素检测呈阴性的MNPs，基于TLR4的缺乏激活和LAL的阴性结果，可能仍然含有选择性激活TLR2的微生物配体。此外，8种MNP测试材料中有5种含有能够激活TLR2和/或TLR4的微生物配体。与LAL法相比，基于tlr4的筛选能有效检测内毒素污染。总的来说，我们发现TLR报告细胞法在检测微生物配体方面比LAL法提供了更广泛的覆盖范围，这在MNP测试材料中似乎非常普遍。

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引用次数: 0

Cisplatin adducts at DNA or RNA do not affect their cellular isolation efficiencies using column-based kits or manual Trizol-based purification methods DNA或RNA上的顺铂加合物使用柱基试剂盒或手动trizol基纯化方法不影响其细胞分离效率。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-06 DOI: 10.1016/j.tiv.2025.106188

Ketaki Sandu, Dirk Theile

In experimental toxicology, evaluating cisplatin adducts at DNA or RNA is often required but can be complicated by methodological aspects. For instance, the cross-linking might affect the nucleic acid isolation efficiencies, which in turn can be influenced by the purification approach. Two cancer cell lines were cisplatin treated and DNA/RNA were isolated using manual Trizol-based protocols or column-based kits. Platinum levels at DNA/RNA were evaluated by atomic absorption spectroscopy.

For RNA, the manual purification yielded higher concentrations than the kit for both cisplatin-treated and non-treated samples. RNA platination was identical for both isolation approaches. For DNA, the manual method can yield lower concentrations from cisplatin-treated cells, likely reflecting diminished solubility of cross-linked DNA. DNA platination levels again were identical with both isolation methods.

Because DNA isolation is less efficient than RNA isolation, platinum-DNA adduct quantification is difficult when DNA yields are low or cells were exposed to low cisplatin concentrations. However, DNA platination can be estimated by the platination degree of RNA because platination of both nucleic acids agreed well and RNA was always isolated very efficiently.

In summary: First, manual purification and column-based kits can yield unequal nucleic acid concentrations, and RNA is more efficiently purified than DNA. Second, column-based kits remain practical because platination does not affect isolation efficiency. Third, when DNA platination is not quantifiable (low yield, small sample volumes), RNA platination is a good proxy.

在实验毒理学中，通常需要评估DNA或RNA上的顺铂加合物，但可能因方法学方面而复杂化。例如，交联可能会影响核酸分离效率，而核酸分离效率又会受到纯化方法的影响。顺铂处理两种癌细胞系，使用手动trizol协议或柱基试剂盒分离DNA/RNA。用原子吸收光谱法测定DNA/RNA中的铂含量。对于RNA，无论是顺铂处理的样本还是未处理的样本，人工纯化的RNA浓度都高于试剂盒。两种分离方法的RNA镀化是相同的。对于DNA，手工方法可以从顺铂处理的细胞中产生较低的浓度，可能反映了交联DNA的溶解度降低。DNA铂化水平再次与两种分离方法相同。由于DNA分离比RNA分离效率低，当DNA产量低或细胞暴露于低顺铂浓度时，铂-DNA加合物的定量是困难的。而DNA的铂化程度可以通过RNA的铂化程度来判断，因为两种核酸的铂化程度一致，RNA的分离效率很高。综上所述：首先，人工纯化和柱式试剂盒可能产生不相等的核酸浓度，RNA的纯化效率高于DNA。其次，基于柱的试剂盒仍然是实用的，因为镀化不会影响分离效率。第三，当DNA铂化无法量化（产量低、样品体积小）时，RNA铂化是一个很好的替代方法。

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引用次数: 0

Optimisation of ultrafine particle exposure in an alveolar tri-culture model at the air-liquid interface 在空气-液体界面的肺泡三培养模型中超细颗粒暴露的优化。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-05 DOI: 10.1016/j.tiv.2025.106189

Anna-Katharina Hensel , Henri Hakkarainen , Mo Yang , Jingwen Huang , Laura Mussalo , Claire Fayad , Katja Kanninen , Pasi Jalava

Air-liquid interface (ALI) systems have emerged as a physiologically relevant in vitro platform for evaluating the toxicological impact and potential health effects of airborne pollutants. When utilizing collected ultrafine particles (UFPs), application volume and liquid type are critical parameters. Using a smaller volume of liquid for the cell exposure results in a heterogeneous distribution of UFPs across the cell monolayer, whereas application of a sufficient volume optimises even UFP distribution. A buffered solution for UFP administration minimises potential side effects and unravels dose-dependent effects in toxicological endpoints. However, standardised exposure methodologies limit reproducibility and comparability across studies. Therefore, we propose a refined manual exposure technique of suspended airborne pollutants in an adequate exposure volume that bridges the gap between conventional submerged cultures and ALI systems. Our model uses cell culture inserts with A549 epithelial cells, THP-1 macrophages, and EA.hy926 endothelial cells to mimic the in vivo alveolar barrier within the lungs. This approach offers a balance of experimental reproducibility whilst addressing the current challenges of standardisation and feasibility in exposure studies with manual UFP exposure. In conjunction with existing aerosol ALI continuous flow exposure systems, our studies are advancing translational in vitro evaluations, aligning with the 3R principle.

气液界面（ALI）系统已成为评估空气污染物的毒理学影响和潜在健康影响的生理相关体外平台。在利用收集的超细颗粒（ufp）时，应用体积和液体类型是关键参数。使用较小体积的液体进行细胞暴露会导致UFP在细胞单层上的不均匀分布，而使用足够体积的液体则会优化UFP的均匀分布。UFP管理的缓冲解决方案最大限度地减少潜在的副作用和解开毒理学终点的剂量依赖效应。然而，标准化暴露方法限制了研究的可重复性和可比性。因此，我们提出了一种改进的人工暴露技术，在适当的暴露量下对悬浮空气污染物进行暴露，以弥补传统的浸入式培养和ALI系统之间的差距。我们的模型使用含有A549上皮细胞、THP-1巨噬细胞和EA.hy926内皮细胞的细胞培养插入物来模拟肺内的肺泡屏障。这种方法提供了实验可重复性的平衡，同时解决了目前手工UFP暴露研究中标准化和可行性的挑战。结合现有的气溶胶ALI连续流动暴露系统，我们的研究正在推进转化体外评估，与3R原则保持一致。

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引用次数: 0

Alternatives to animal testing in cosmetic products: A patent applications review and future perspectives 化妆品动物试验的替代方案：专利申请审查和未来展望。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-03 DOI: 10.1016/j.tiv.2025.106187

Carine Cassimiro Cedrola, Clara Cassimiro Cedrola, Ana Cláudia Chagas de Paula, Juliana de Carvalho da Costa, Fernanda Maria Pinto Vilela

Ethical concerns, high costs, and scientific limitations associated with animal testing have accelerated the search for alternative methods to evaluate the safety and efficacy of topical cosmetic formulations. This study provides a comprehensive analysis of the global patent landscape related to non-animal testing approaches for skin care products, focusing on filings from January 2015 to March 2025, indexed in the Espacenet database. From 470 patent applications initially screened, 23 met the predefined inclusion and exclusion criteria and were selected for in-depth analysis. Key innovations include 3D epidermal models featuring melanocytes, hair follicles, and sebaceous glands; advanced microfluidic chips, and enzyme-based chemical toxicity assays. Although supported by regulatory frameworks, challenges persist regarding standardization, reproducibility, and the ethical sourcing of human tissue. This patent application review reveals a clear shift toward advanced 3D models and organ-on-a-chip technologies that better replicate the complexity of human skin physiology. The trends observed indicate that alternative methods to animal testing are not only an ethical necessity but are also becoming a technological reality, offering more predictive, reliable, and efficient strategies for safety assessment.

与动物试验相关的伦理问题、高成本和科学限制加速了对替代方法的探索，以评估局部化妆品配方的安全性和有效性。本研究对全球护肤品非动物试验方法专利格局进行了全面分析，重点关注2015年1月至2025年3月的申请，并在Espacenet数据库中检索。从最初筛选的470项专利申请中，有23项符合预定义的纳入和排除标准，并被选中进行深入分析。主要创新包括以黑色素细胞、毛囊和皮脂腺为特征的3D表皮模型；先进的微流控芯片，以及基于酶的化学毒性分析。尽管得到了监管框架的支持，但在人体组织的标准化、可重复性和道德来源方面仍然存在挑战。这项专利申请审查揭示了向先进的3D模型和器官芯片技术的明显转变，这些技术可以更好地复制人类皮肤生理学的复杂性。观察到的趋势表明，动物试验的替代方法不仅在伦理上是必要的，而且正在成为一种技术现实，为安全性评估提供更有预测性、更可靠和更有效的策略。

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引用次数: 0

Evaluating cannabidiol-induced liver injury with and without valproate using a three-dimensional human hepatocyte spheroid model. 使用三维人肝细胞球体模型评估大麻二酚诱导的有丙戊酸和无丙戊酸肝损伤。

IF 2.7 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-12-01 Epub Date: 2025-08-05 DOI: 10.1016/j.tiv.2025.106126

Jessica L Beers, Shalon L Harvey, Raeanne M Lanphier, Blake R Rushing, Susan L McRitchie, Susan J Sumner, Klarissa D Jackson

Cannabidiol (CBD) and valproate (VPA) are anti-epileptic medications commonly co-prescribed to treat seizures due to Lennox-Gastaut syndrome, Dravet syndrome, and tuberous sclerosis complex in children. Clinical trial data have demonstrated that CBD carries a risk for severe hepatotoxicity that is greatly increased when prescribed with VPA through an unknown mechanism. The aim of this study was to investigate CBD-induced liver injury in combination with VPA using an in vitro liver model. Three-dimensional human hepatocyte spheroids are an emerging in vitro system that allows investigation of long-term toxicity. Spheroids derived from primary human hepatocytes were treated with vehicle control, 2-200 μM CBD, 0.5-20 mM VPA, CBD + VPA, and 0.1-10 mM acetaminophen (positive control). After 24 h, 8 days, and 15 days of exposure, spheroids were analyzed for ATP depletion, urea production, and CBD and VPA metabolite generation. Untargeted metabolomic analysis was also conducted. A delayed-onset, dose-dependent hepatotoxicity was observed in spheroids exposed to each drug treatment compared to vehicle control. This study is the first to recapitulate the hepatotoxic drug interaction of CBD and VPA in vitro and demonstrates the utility of human hepatocyte spheroids for toxicity studies. Future work is needed to examine mechanisms of CBD-induced hepatotoxicity with VPA.

大麻二酚（CBD）和丙戊酸酯（VPA）是抗癫痫药物，通常共同用于治疗儿童Lennox-Gastaut综合征，Dravet综合征和结节性硬化症引起的癫痫发作。临床试验数据表明，CBD具有严重肝毒性的风险，当通过未知机制与VPA一起使用时，这种风险会大大增加。本研究的目的是通过体外肝模型研究cbd联合VPA诱导的肝损伤。三维人肝细胞球体是一个新兴的体外系统，允许研究长期毒性。来源于人原代肝细胞的球体分别用对照、2-200 μM CBD、0.5-20 mM VPA、CBD + VPA和0.1-10 mM对乙酰氨基酚（阳性对照）处理。在暴露24小时、8天和15天后，对球体进行ATP消耗、尿素生成、CBD和VPA代谢物生成的分析。还进行了非靶向代谢组学分析。与对照组相比，暴露于每种药物治疗的球体中观察到延迟发作，剂量依赖性肝毒性。这项研究首次概述了CBD和VPA在体外的肝毒性药物相互作用，并证明了人类肝细胞球体在毒性研究中的实用性。未来的工作需要研究cbd诱导的VPA肝毒性的机制。

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