Toxicology in Vitro最新文献_第8页

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。

{"title":"The protective mechanisms of glycyrrhizic acid for deoxynivalenol-induced toxicity in rabbit corneal stroma","authors":"Chunlan Liang , Xuewei Xiong , Qi Shi , Qingqing Li , Guocheng Yu , Jingxiang Zhong , Lian Liu","doi":"10.1016/j.tiv.2025.106104","DOIUrl":"10.1016/j.tiv.2025.106104","url":null,"abstract":"<div><h3>Objectives</h3><div>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.</div></div><div><h3>Methods</h3><div>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.</div></div><div><h3>Results</h3><div>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.</div></div><div><h3>Conclusions</h3><div>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.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106104"},"PeriodicalIF":2.6,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568074","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}

引用次数: 0

Corrigendum to “Assessing the safety of midazolam: A comprehensive analysis of adverse events from FAERS” [Toxicology in vitro 105 (2025) 106023] “咪达唑仑的安全性评估：FAERS不良事件的综合分析”的更正[体外毒理学105 (2025)106023]

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-07-01 DOI: 10.1016/j.tiv.2025.106103

Jieyuan Chen , Zhaojun Wang , Li Wei , Songsong Mao

引用次数: 0

Dodecyldimethylamine oxide may induce lysosomal-associated cell death: Potential lipotoxicity 十二烷基二甲胺氧化物可能诱导溶酶体相关细胞死亡：潜在的脂肪毒性。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-06-24 DOI: 10.1016/j.tiv.2025.106102

Eun-Jung Park , Minseok Seo , Wonkyun Jung , Hyosun Choi , Ji Ae Lee

Dodecyldimethylamine oxide (DDAO) is one of the most commonly used chemicals in household products, including spray types. We identified the potential toxicity mechanism using human bronchial epithelial cells. After 6 h of exposure, DDAO decreased cell viability (non-apoptotic cell death) and formed numerous small artificial vacuoles in the cytoplasm. Cellular impedance decreased rapidly following exposure to DDAO, and condensation of the inner mitochondrial membrane, an increased level of cytochrome C, and a decrease in ATP production and mitochondrial volume were observed in DDAO-treated cells, affecting the level of second messengers. Impaired intracellular components, including lamellar bodies, were also found within the mitochondria or vacuoles. While the secretion of chemotactic cytokines decreased, the levels of IL-6, IL-11, MMP-1, and MMP-3 increased. Furthermore, DDAO enhanced the expression of the LAMP-2 protein and notably increased the volume of acidic compartments. More importantly, the expression of lipid metabolism-related genes was most affected following DDAO treatment. Considering that the volume of acidic compartments decreased rapidly at a concentration of 40 μg/mL, we suggest that DDAO may induce lysosomal-associated cell death (possible lipotoxicity) through structural damage to the cell (mitochondria) membrane, and that lysosomal membrane proteins may contribute to protecting cells from DDAO-induced impacts.

十二烷基二甲胺氧化物（DDAO）是家用产品中最常用的化学品之一，包括喷雾类型。我们利用人支气管上皮细胞确定了潜在的毒性机制。暴露6 h后，DDAO降低细胞活力（非凋亡细胞死亡），并在细胞质中形成许多小的人工液泡。DDAO处理后，细胞阻抗迅速下降，线粒体内膜凝结，细胞色素C水平升高，ATP产量和线粒体体积减少，影响第二信使的水平。在线粒体或液泡内也发现了受损的细胞内成分，包括板层体。趋化因子分泌减少，IL-6、IL-11、MMP-1、MMP-3水平升高。此外，DDAO增强了LAMP-2蛋白的表达，并显著增加了酸性区室的体积。更重要的是，DDAO治疗对脂质代谢相关基因的表达影响最大。考虑到酸性区室的体积在浓度为40 μg/mL时迅速减少，我们认为DDAO可能通过对细胞（线粒体）膜的结构损伤诱导溶酶体相关细胞死亡（可能是脂毒性），溶酶体膜蛋白可能有助于保护细胞免受DDAO诱导的影响。

{"title":"Dodecyldimethylamine oxide may induce lysosomal-associated cell death: Potential lipotoxicity","authors":"Eun-Jung Park , Minseok Seo , Wonkyun Jung , Hyosun Choi , Ji Ae Lee","doi":"10.1016/j.tiv.2025.106102","DOIUrl":"10.1016/j.tiv.2025.106102","url":null,"abstract":"<div><div>Dodecyldimethylamine oxide (DDAO) is one of the most commonly used chemicals in household products, including spray types. We identified the potential toxicity mechanism using human bronchial epithelial cells. After 6 h of exposure, DDAO decreased cell viability (non-apoptotic cell death) and formed numerous small artificial vacuoles in the cytoplasm. Cellular impedance decreased rapidly following exposure to DDAO, and condensation of the inner mitochondrial membrane, an increased level of cytochrome C, and a decrease in ATP production and mitochondrial volume were observed in DDAO-treated cells, affecting the level of second messengers. Impaired intracellular components, including lamellar bodies, were also found within the mitochondria or vacuoles. While the secretion of chemotactic cytokines decreased, the levels of IL-6, IL-11, MMP-1, and MMP-3 increased. Furthermore, DDAO enhanced the expression of the LAMP-2 protein and notably increased the volume of acidic compartments. More importantly, the expression of lipid metabolism-related genes was most affected following DDAO treatment. Considering that the volume of acidic compartments decreased rapidly at a concentration of 40 μg/mL, we suggest that DDAO may induce lysosomal-associated cell death (possible lipotoxicity) through structural damage to the cell (mitochondria) membrane, and that lysosomal membrane proteins may contribute to protecting cells from DDAO-induced impacts.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106102"},"PeriodicalIF":2.6,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144509379","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}

引用次数: 0

In vitro evaluation of antimicrobial food packaging: Assessing its inflammatory and sensitization potential 抗菌食品包装的体外评价：评估其炎症和致敏潜力

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-06-15 DOI: 10.1016/j.tiv.2025.106083

Franziska Trodtfeld, Cornelia Wiegand

Skin sensitization is a key endpoint in chemical safety assessment for cosmetics and personal care products and is increasingly suggested for consideration in the evaluation of packaging materials, especially food packaging materials. Keratinocyte activation, involving oxidative stress, cytokine release, and the engagement of stress-related signaling pathways, represents an early and critical event in this process. In this study, we evaluated the responses of primary keratinocytes (PK), epidermis equivalents exposed to solutions (Epi-Sol), and coatings (Epi-Coat) to known skin sensitizers and antimicrobial agents included in food packaging.

Cytokine measurements (IL-1α, IL-6, IL-8, IL-18), gene expression profiling of stress and immune-related genes (HMOX1, MMP10, DUSP1, HSP90AA1, ICAM1, IL4R, IL23A, IL33), and histological analyses were performed. PK models exhibited the strongest and most diverse responses, with consistent HMOX1 upregulation and robust inflammatory signaling. Epi-Sol models exhibited moderate responses, whereas Epi-Coat models showed lower reactivity, emphasizing the impact of both exposure method and model selection. Nanoparticles (Ag-NPs, ZnO-NPs) triggered exposure-dependent effects, with coatings limiting inflammatory signaling and tissue penetration. Histological changes confirmed barrier disruption and keratinocyte hyperproliferation in models exposed to stronger sensitizers.

Overall, these findings demonstrate that no single biomarker or model is sufficient for accurate safety assessment and support the use of multi-test, multi-endpoint evaluation strategies — particularly for emerging applications like food-contact materials — to ensure reliable hazard identification and material safety evaluation.

皮肤致敏性是化妆品和个人护理用品化学安全评价的关键指标，在包装材料尤其是食品包装材料的评价中越来越多地被建议考虑。角质形成细胞的激活，包括氧化应激、细胞因子释放和应激相关信号通路的参与，代表了这一过程的早期和关键事件。在这项研究中，我们评估了初代角质形成细胞（PK）、暴露于溶液（Epi-Sol）的表皮当量和涂层（Epi-Coat）对食品包装中已知皮肤致敏剂和抗菌剂的反应。细胞因子测定（IL-1α、IL-6、IL-8、IL-18）、应激和免疫相关基因（HMOX1、MMP10、DUSP1、HSP90AA1、ICAM1、IL4R、IL23A、IL33）的基因表达谱和组织学分析。PK模型表现出最强和最多样化的反应，具有一致的HMOX1上调和强大的炎症信号。Epi-Sol模型表现出中等的反应性，而Epi-Coat模型表现出较低的反应性，强调了暴露方法和模型选择的影响。纳米颗粒（Ag-NPs, ZnO-NPs）引发暴露依赖性效应，其涂层限制了炎症信号和组织渗透。在暴露于强致敏剂的模型中，组织学变化证实了屏障破坏和角化细胞过度增殖。总的来说，这些发现表明，没有单一的生物标志物或模型足以进行准确的安全性评估，并支持使用多测试，多端点评估策略-特别是对于食品接触材料等新兴应用-以确保可靠的危害识别和材料安全性评估。

{"title":"In vitro evaluation of antimicrobial food packaging: Assessing its inflammatory and sensitization potential","authors":"Franziska Trodtfeld, Cornelia Wiegand","doi":"10.1016/j.tiv.2025.106083","DOIUrl":"10.1016/j.tiv.2025.106083","url":null,"abstract":"<div><div>Skin sensitization is a key endpoint in chemical safety assessment for cosmetics and personal care products and is increasingly suggested for consideration in the evaluation of packaging materials, especially food packaging materials. Keratinocyte activation, involving oxidative stress, cytokine release, and the engagement of stress-related signaling pathways, represents an early and critical event in this process. In this study, we evaluated the responses of primary keratinocytes (PK), epidermis equivalents exposed to solutions (Epi-Sol), and coatings (Epi-Coat) to known skin sensitizers and antimicrobial agents included in food packaging.</div><div>Cytokine measurements (IL-1α, IL-6, IL-8, IL-18), gene expression profiling of stress and immune-related genes (<em>HMOX1, MMP10, DUSP1, HSP90AA1, ICAM1, IL4R, IL23A, IL33</em>), and histological analyses were performed. PK models exhibited the strongest and most diverse responses, with consistent <em>HMOX1</em> upregulation and robust inflammatory signaling. Epi-Sol models exhibited moderate responses, whereas Epi-Coat models showed lower reactivity, emphasizing the impact of both exposure method and model selection. Nanoparticles (Ag-NPs, ZnO-NPs) triggered exposure-dependent effects, with coatings limiting inflammatory signaling and tissue penetration. Histological changes confirmed barrier disruption and keratinocyte hyperproliferation in models exposed to stronger sensitizers.</div><div>Overall, these findings demonstrate that no single biomarker or model is sufficient for accurate safety assessment and support the use of multi-test, multi-endpoint evaluation strategies — particularly for emerging applications like food-contact materials — to ensure reliable hazard identification and material safety evaluation.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106083"},"PeriodicalIF":2.6,"publicationDate":"2025-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144312903","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}

引用次数: 0

Corrigendum to “Integrative toxicity assessment of tocotrienol-rich fraction from palm oil using in silico methods and zebrafish embryotoxicity model” [Toxicology in Vitro, 107 (2025) 106062] “利用计算机方法和斑马鱼胚胎毒性模型对棕榈油中富含生育三烯醇的部分进行综合毒性评估”的勘误表[体外毒理学，107 (2025)106062]

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-06-12 DOI: 10.1016/j.tiv.2025.106099

Thenmoly Damodaran , Najib Sani Yahaya , Mohd Nizam Mordi

引用次数: 0

Effects of an environmental chemical mixture on early-stage embryo development: In vitro evidence from human embryonic stem cells 环境化学混合物对早期胚胎发育的影响：来自人类胚胎干细胞的体外证据。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-06-12 DOI: 10.1016/j.tiv.2025.106101

Bai Li , Yihang Kevin Pan , Xiaolei Jin , Hing Man Chan

Environmental chemicals are known risk factors for adverse pregnancy outcomes. However, the effects of environmentally relevant concentrations of chemical mixtures are less studied. This study investigated the effects of a mixture containing 23 chemicals reported in blood samples of Nunavik pregnant women, termed the Nunavik Chemical Mixture (NCM), on embryo development using human embryonic stem cells (hESCs). hESCs were exposed to 0-100X of NCM (X is the sum of geometric mean concentrations of NCM's components) for 24 h and 6 d Cell viability, apoptosis, stress response, cell cycle, cytoskeleton, autophagy, and expression of lineage marker genes/proteins were measured after exposures. NCM decreased cell viability and adhesion, induced apoptosis, disrupted the cell cycle, and altered the expression of cytoskeleton, autophagy proteins, and lineage marker genes/proteins in a dose-dependent manner. These results suggested that NCM affected embryo development, leading to potential adverse pregnancy outcomes if it occurs in vivo. Moreover, the effects caused by NCM were different from those caused by the same doses of MeHg alone that we previously found, indicating potential interactions among components within the mixture. Our results highlight the importance of considering the potential combined effects of chemical mixtures when assessing health risks in populations exposed to various environmental chemicals.

众所周知，环境化学物质是导致不良妊娠结果的危险因素。然而，对与环境有关的化学混合物浓度的影响的研究较少。这项研究调查了努纳维克孕妇血液样本中含有23种化学物质的混合物，称为努纳维克化学混合物（NCM），对人类胚胎干细胞（hESCs）胚胎发育的影响。将hESCs暴露于0-100倍的NCM （X为NCM各组分几何平均浓度之和）中24 h和6 d后，检测细胞活力、凋亡、应激反应、细胞周期、细胞骨架、自噬和谱系标记基因/蛋白的表达。NCM降低细胞活力和粘附，诱导细胞凋亡，破坏细胞周期，并以剂量依赖的方式改变细胞骨架、自噬蛋白和谱系标记基因/蛋白的表达。这些结果表明，NCM影响胚胎发育，如果发生在体内，可能会导致潜在的不良妊娠结局。此外，NCM引起的影响与我们之前发现的相同剂量的MeHg单独引起的影响不同，这表明混合物中成分之间可能存在相互作用。我们的研究结果强调了在评估暴露于各种环境化学品的人群的健康风险时，考虑化学混合物的潜在综合影响的重要性。

{"title":"Effects of an environmental chemical mixture on early-stage embryo development: In vitro evidence from human embryonic stem cells","authors":"Bai Li , Yihang Kevin Pan , Xiaolei Jin , Hing Man Chan","doi":"10.1016/j.tiv.2025.106101","DOIUrl":"10.1016/j.tiv.2025.106101","url":null,"abstract":"<div><div>Environmental chemicals are known risk factors for adverse pregnancy outcomes. However, the effects of environmentally relevant concentrations of chemical mixtures are less studied. This study investigated the effects of a mixture containing 23 chemicals reported in blood samples of Nunavik pregnant women, termed the Nunavik Chemical Mixture (NCM), on embryo development using human embryonic stem cells (hESCs). hESCs were exposed to 0-100X of NCM (X is the sum of geometric mean concentrations of NCM's components) for 24 h and 6 d Cell viability, apoptosis, stress response, cell cycle, cytoskeleton, autophagy, and expression of lineage marker genes/proteins were measured after exposures. NCM decreased cell viability and adhesion, induced apoptosis, disrupted the cell cycle, and altered the expression of cytoskeleton, autophagy proteins, and lineage marker genes/proteins in a dose-dependent manner. These results suggested that NCM affected embryo development, leading to potential adverse pregnancy outcomes if it occurs <em>in vivo</em>. Moreover, the effects caused by NCM were different from those caused by the same doses of MeHg alone that we previously found, indicating potential interactions among components within the mixture. Our results highlight the importance of considering the potential combined effects of chemical mixtures when assessing health risks in populations exposed to various environmental chemicals.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106101"},"PeriodicalIF":2.6,"publicationDate":"2025-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144295312","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}

引用次数: 0

CiPA-qualified human iPSC-derived cardiomyocytes: A new frontier in toxicity testing by evaluating drug-induced arrhythmias cipa合格的人ipsc衍生心肌细胞：通过评估药物性心律失常进行毒性测试的新前沿。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-06-05 DOI: 10.1016/j.tiv.2025.106100

Vijay Bhaskar Reddy Konala , Rutuja Kuhikar , Shruti More , Matthias Gossmann , Bettina Lickiss , Peter Linder , Jaganmay Sarkar , Paresh Bhanushali , Amit Khanna

Drug-induced arrhythmias remain a significant challenge in drug development, often leading to serious cardiovascular complications and the withdrawal of approved drugs from the market. The Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative aims to enhance cardiac safety assessment by leveraging human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). In this study, we evaluated the effects of 28 drugs on a well-characterized hiPSC-CMs (YBLiCardio, Yashraj Biotechnology Ltd., Mumbai, India) using Electric Field Potential (EFP) measurements. The CardioExcyte 96 system recorded extracellular signals from 96 wells, functioning similarly to microelectrode arrays. Each drug was tested at four concentrations, and the effects were analyzed based on dynamic changes in beat patterns, with QT prolongation assessed by measuring the interval between the sodium spike and T-wave. Our results demonstrated that YBLiCardio cells responded to all drugs in line with the findings from the HESI CiPA study. Notably, droperidol (173 %) and domperidone (182 %), originally classified as intermediate-risk compounds, were identified as high-risk in our model, consistent with previous findings by Nguyen et al. (2017). Additionally, YBLiCardio showed enhanced predictive accuracy for chlorpromazine. These findings highlight the potential of hiPSC-CMs for proarrhythmia risk assessment within the CiPA framework, complementing ion channel data and in silico modeling approaches. Overall, YBLiCardio provides a robust and physiologically relevant platform for predicting cardiotoxicity, supporting safer and more efficient pre-clinical drug discovery & development.

药物性心律失常仍然是药物开发中的一个重大挑战，经常导致严重的心血管并发症和已批准的药物退出市场。综合体外心律失常检测（CiPA）计划旨在通过利用人类诱导的多能干细胞来源的心肌细胞（hiPSC-CMs）来加强心脏安全性评估。在本研究中，我们利用电场电位（EFP）测量方法评估了28种药物对具有良好特征的hiPSC-CMs （YBLiCardio, Yashraj Biotechnology Ltd., Mumbai, India）的影响。CardioExcyte 96系统记录了来自96个孔的细胞外信号，其功能类似于微电极阵列。每种药物在四种浓度下进行测试，并根据心跳模式的动态变化分析效果，通过测量钠峰和t波之间的间隔来评估QT延长。我们的研究结果表明，YBLiCardio细胞对所有药物都有反应，这与HESI CiPA研究的结果一致。值得注意的是，原本被归类为中等风险化合物的氟哌啶醇（173 %）和多潘立酮（182 %）在我们的模型中被确定为高风险，这与Nguyen等人（2017）之前的发现一致。此外，YBLiCardio对氯丙嗪的预测准确性也有所提高。这些发现强调了hiPSC-CMs在CiPA框架内用于心律失常风险评估的潜力，补充了离子通道数据和计算机建模方法。总的来说，YBLiCardio为预测心脏毒性提供了一个强大的生理学相关平台，支持更安全、更有效的临床前药物发现和开发。

{"title":"CiPA-qualified human iPSC-derived cardiomyocytes: A new frontier in toxicity testing by evaluating drug-induced arrhythmias","authors":"Vijay Bhaskar Reddy Konala , Rutuja Kuhikar , Shruti More , Matthias Gossmann , Bettina Lickiss , Peter Linder , Jaganmay Sarkar , Paresh Bhanushali , Amit Khanna","doi":"10.1016/j.tiv.2025.106100","DOIUrl":"10.1016/j.tiv.2025.106100","url":null,"abstract":"<div><div>Drug-induced arrhythmias remain a significant challenge in drug development, often leading to serious cardiovascular complications and the withdrawal of approved drugs from the market. The Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative aims to enhance cardiac safety assessment by leveraging human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). In this study, we evaluated the effects of 28 drugs on a well-characterized hiPSC-CMs (YBLiCardio, Yashraj Biotechnology Ltd., Mumbai, India) using Electric Field Potential (EFP) measurements. The CardioExcyte 96 system recorded extracellular signals from 96 wells, functioning similarly to microelectrode arrays. Each drug was tested at four concentrations, and the effects were analyzed based on dynamic changes in beat patterns, with QT prolongation assessed by measuring the interval between the sodium spike and T-wave. Our results demonstrated that YBLiCardio cells responded to all drugs in line with the findings from the HESI CiPA study. Notably, droperidol (173 %) and domperidone (182 %), originally classified as intermediate-risk compounds, were identified as high-risk in our model, consistent with previous findings by Nguyen et al. (2017). Additionally, YBLiCardio showed enhanced predictive accuracy for chlorpromazine. These findings highlight the potential of hiPSC-CMs for proarrhythmia risk assessment within the CiPA framework, complementing ion channel data and in silico modeling approaches. Overall, YBLiCardio provides a robust and physiologically relevant platform for predicting cardiotoxicity, supporting safer and more efficient pre-clinical drug discovery & development.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106100"},"PeriodicalIF":2.6,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144250803","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}

引用次数: 0

Chronic exposure to palmitic acid-induced adipocyte hypertrophy and altered batokine gene expression in T37i brown adipocytes 慢性暴露于棕榈酸诱导的脂肪细胞肥大和T37i棕色脂肪细胞中细胞因子基因表达的改变。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-05-29 DOI: 10.1016/j.tiv.2025.106097

Khanyisani Ziqubu , Phiwayinkosi V. Dludla , Sithandiwe E. Mazibuko-Mbeje

Enlargement of adipose tissue through hypertrophy is a key hallmark of obesity. Our previous study demonstrated that chronic obesity induces brown adipose tissue hypertrophy and altered batokine gene expression patterns in vivo. The present study further explored and verified the pathophysiological and molecular changes implicated in brown adipocyte hypertrophy by exposing T37i cells to 0.25, 0.5, 0.75, and 1 mM of palmitic acid for 48 h. The results showed that palmitic acid-induced intracellular lipid accumulation and lipolysis. Gene expression analysis demonstrated that palmitic acid downregulated genes responsible for glucose and lipid metabolism, such as AdipoQ and PIk3r1, while upregulating Cpt1A, a mitochondrial fatty acid transporter, and Tnf-α, a pro-inflammatory cytokine. Moreover, palmitic acid downregulated brown adipocyte transcriptional factors and thermogenic markers, including Prdm16, Pparg, Cidea, Dio2, Sirt1, and Ucp1. Gene expression of batokines involved in regulating substrate metabolism (Fgf21), angiogenesis (Nrg4 and VegfA), and immune cell recruitment (Metrnl, Gdf15, and Cxcl14) were altered by palmitic acid. This data has demonstrated that palmitic acid contributes to the hypertrophy and whitening of brown adipocytes by inhibiting brown adipocyte differentiation and altering batokines expression patterns.

脂肪组织因肥大而增大是肥胖的一个重要标志。我们之前的研究表明，慢性肥胖诱导体内棕色脂肪组织肥大和改变细胞因子基因表达模式。本研究通过将T37i细胞暴露于0.25、0.5、0.75和1 mM的棕榈酸中48 h，进一步探索和验证了褐色脂肪细胞肥大的病理生理和分子变化。结果表明，棕榈酸可诱导细胞内脂质积累和脂质分解。基因表达分析显示，棕榈酸下调糖脂代谢相关基因AdipoQ和PIk3r1，上调线粒体脂肪酸转运体Cpt1A和促炎细胞因子Tnf-α。此外，棕榈酸下调棕色脂肪细胞转录因子和产热标志物，包括Prdm16、Pparg、Cidea、Dio2、Sirt1和Ucp1。棕榈酸改变了调节底物代谢（Fgf21）、血管生成（Nrg4和VegfA）和免疫细胞募集（Metrnl、Gdf15和Cxcl14）的细胞因子的基因表达模式。这些数据表明，棕榈酸通过抑制棕色脂肪细胞分化和改变细胞因子表达模式，有助于棕色脂肪细胞的肥大和增白。

{"title":"Chronic exposure to palmitic acid-induced adipocyte hypertrophy and altered batokine gene expression in T37i brown adipocytes","authors":"Khanyisani Ziqubu , Phiwayinkosi V. Dludla , Sithandiwe E. Mazibuko-Mbeje","doi":"10.1016/j.tiv.2025.106097","DOIUrl":"10.1016/j.tiv.2025.106097","url":null,"abstract":"<div><div>Enlargement of adipose tissue through hypertrophy is a key hallmark of obesity. Our previous study demonstrated that chronic obesity induces brown adipose tissue hypertrophy and altered batokine gene expression patterns <em>in vivo</em>. The present study further explored and verified the pathophysiological and molecular changes implicated in brown adipocyte hypertrophy by exposing T37i cells to 0.25, 0.5, 0.75, and 1 mM of palmitic acid for 48 h. The results showed that palmitic acid-induced intracellular lipid accumulation and lipolysis. Gene expression analysis demonstrated that palmitic acid downregulated genes responsible for glucose and lipid metabolism, such as <em>AdipoQ</em> and <em>PIk3r1</em>, while upregulating <em>Cpt1A</em>, a mitochondrial fatty acid transporter, and <em>Tnf-α</em>, a pro-inflammatory cytokine. Moreover, palmitic acid downregulated brown adipocyte transcriptional factors and thermogenic markers, including <em>Prdm16</em>, <em>Pparg</em>, <em>Cidea</em>, <em>Dio2</em>, <em>Sirt1,</em> and <em>Ucp1</em>. Gene expression of batokines involved in regulating substrate metabolism (<em>Fgf21</em>), angiogenesis (<em>Nrg4</em> and <em>VegfA</em>), and immune cell recruitment (<em>Metrnl</em>, <em>Gdf15,</em> and <em>Cxcl14</em>) were altered by palmitic acid. This data has demonstrated that palmitic acid contributes to the hypertrophy and whitening of brown adipocytes by inhibiting brown adipocyte differentiation and altering batokines expression patterns.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106097"},"PeriodicalIF":2.6,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144192530","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}

引用次数: 0

Toxicoproteomic study of fipronil in SH-SY5Y cells reveals induction of endoplasmic reticulum stress and necrotic cell death as neurodegenerative mechanisms 氟虫腈在SH-SY5Y细胞中的毒性蛋白质组学研究揭示了内质网应激和坏死细胞死亡是神经退行性机制

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-05-29 DOI: 10.1016/j.tiv.2025.106098

Theetat Ruangjaroon , N. Monique Paricharttanakul , Daranee Chokchaichamnankit , Chantragan Srisomsap , Kriengsak Lirdprapamongkol , Jisnuson Svasti

Exposure to pesticides has been considered as a risk factor for developing neurodegenerative diseases. The increasing use of fipronil, a phenylpyrazole insecticide, poses a risk to human health. This study aims to use toxicoproteomics for exploring neurodegenerative mechanism of fipronil in SH-SY5Y human neuroblastoma cells. In this study, fipronil at sub-cytotoxic and cytotoxic concentrations (43 and 78 μM) caused increases in superoxide level from 3 to 48 h after treatment, while intracellular glutathione level was decreased at 48 h. Neurite outgrowth of the cells was impaired by fipronil at both concentrations, while significant increase of cell death via apoptosis and necrosis modes were observed with fipronil at cytotoxic concentration. Pretreatment with antioxidant N-acetylcysteine (NAC) effectively relieved impairment of neurite outgrowth and induction of cell death by fipronil. Proteomic analysis showed that expression of proteins involving endoplasmic reticulum (ER) stress and unfolded protein responses were predominantly affected by fipronil. Immunoblotting confirmed the increased expression of ER stress markers, GRP78/BiP (78 kDa glucose-regulated protein/Binding immunoglobulin protein) and PDI (protein disulfide isomerase), in fipronil-treated cells. Improved understanding of the neurotoxic mechanism of fipronil may help in developing a strategy for reducing risk of neurodegenerative development from intense and prolonged use of fipronil.

接触杀虫剂被认为是发生神经退行性疾病的一个危险因素。越来越多地使用苯基吡唑类杀虫剂氟虫腈，对人类健康构成威胁。本研究旨在利用毒理学蛋白质组学方法探讨氟虫腈对SH-SY5Y人神经母细胞瘤细胞的神经退行性作用机制。在本研究中，亚细胞毒性和细胞毒性浓度（43 μM和78 μM）的氟虫腈在处理后3 ~ 48 h内引起超氧化物水平升高，而细胞内谷胱甘肽水平在48 h内降低。两种浓度的氟虫腈均损害细胞的神经突生长，同时细胞毒性浓度的氟虫腈显著增加细胞凋亡和坏死模式的死亡。抗氧化剂n -乙酰半胱氨酸（NAC）预处理可有效缓解氟虫腈诱导的神经突生长损伤和细胞死亡。蛋白质组学分析显示，参与内质网应激和未折叠蛋白反应的蛋白质表达主要受到氟虫腈的影响。免疫印迹证实，在氟虫腈处理的细胞中，内质网应激标志物GRP78/BiP （78 kDa葡萄糖调节蛋白/结合免疫球蛋白蛋白）和PDI（蛋白二硫异构酶）的表达增加。提高对氟虫腈神经毒性机制的了解可能有助于制定一种策略，以减少因长期高强度使用氟虫腈而导致的神经退行性发展的风险。

{"title":"Toxicoproteomic study of fipronil in SH-SY5Y cells reveals induction of endoplasmic reticulum stress and necrotic cell death as neurodegenerative mechanisms","authors":"Theetat Ruangjaroon , N. Monique Paricharttanakul , Daranee Chokchaichamnankit , Chantragan Srisomsap , Kriengsak Lirdprapamongkol , Jisnuson Svasti","doi":"10.1016/j.tiv.2025.106098","DOIUrl":"10.1016/j.tiv.2025.106098","url":null,"abstract":"<div><div>Exposure to pesticides has been considered as a risk factor for developing neurodegenerative diseases. The increasing use of fipronil, a phenylpyrazole insecticide, poses a risk to human health. This study aims to use toxicoproteomics for exploring neurodegenerative mechanism of fipronil in SH-SY5Y human neuroblastoma cells. In this study, fipronil at sub-cytotoxic and cytotoxic concentrations (43 and 78 μM) caused increases in superoxide level from 3 to 48 h after treatment, while intracellular glutathione level was decreased at 48 h. Neurite outgrowth of the cells was impaired by fipronil at both concentrations, while significant increase of cell death via apoptosis and necrosis modes were observed with fipronil at cytotoxic concentration. Pretreatment with antioxidant <em>N</em>-acetylcysteine (NAC) effectively relieved impairment of neurite outgrowth and induction of cell death by fipronil. Proteomic analysis showed that expression of proteins involving endoplasmic reticulum (ER) stress and unfolded protein responses were predominantly affected by fipronil. Immunoblotting confirmed the increased expression of ER stress markers, GRP78/BiP (78 kDa glucose-regulated protein/Binding immunoglobulin protein) and PDI (protein disulfide isomerase), in fipronil-treated cells. Improved understanding of the neurotoxic mechanism of fipronil may help in developing a strategy for reducing risk of neurodegenerative development from intense and prolonged use of fipronil.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106098"},"PeriodicalIF":2.6,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144178811","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}

引用次数: 0

Use of centromeric probe to identify micronuclei origin and its advantages in genetic toxicology studies 利用着丝粒探针鉴定微核起源及其在遗传毒理学研究中的优势

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2025-05-26 DOI: 10.1016/j.tiv.2025.106087

G. Tamizh Selvan , P. Venkatachalam

Fluorescence in situ hybridization (FISH) and karyotyping have long been considered essential for chromosomal examination to analyse the stable chromosome aberrations such as translocations after exposure to genotoxic agents. To improve speed of chromosomal analysis in genetic toxicology investigations, we employed the cytokinesis-block micronucleus (CBMN) assay with human pan-centromeric probes, to measure chromosome segregation errors and to differentiate mechanism of action of genotoxic agents. Whole-blood collected from healthy volunteers were exposed to X-rays, bleomycin (BLM), Colchicine (COL) and Mitomycin-C (MMC), arrested at cytokinesis stage and processed for conventional giemsa staining and pan-centromeric FISH to analyse the composition of MN to differentiate between aneugen and clastogen effects. One-way ANOVA with Tukey's multiple comparison test exhibited a significant (p < 0.001) increase in the MN observed using giemsa stained and centromere FISH binucleated cells in all those genotoxic agents when compared to its unexposed lymphocytes cultures. The CBMN with centromere FISH, predominantly shows MNCN^-ve cells in lymphocytes exposed to X-rays (67.4 % and 83.3 %), BLM (79.4 % and 79.2 %) and MMC (81.5 % and 73.2) while, COL treatment resulted in significantly (p < 0.05) higher MNCN^+ve cells (79.6 % and 82.4 %) for the different concentrations of those agents. Centromere FISH enables to identify the origin of MN induced by different genotoxic agents. Thus, a significant development in genetic toxicology is heralded by the incorporation of centromeric probe in modern analytical workflows.

长期以来，荧光原位杂交（FISH）和染色体核型分析一直被认为是染色体检查中分析遗传毒性物质暴露后染色体易位等稳定畸变的必要手段。为了提高遗传毒理学研究中染色体分析的速度，我们采用细胞分裂-阻断微核（CBMN）实验，用人类泛着丝探针测量染色体分离错误，并区分遗传毒性药物的作用机制。从健康志愿者收集的全血暴露于x射线，博来霉素（BLM），秋水仙碱（COL）和丝裂霉素c (MMC)，在细胞分裂阶段捕获，并进行常规吉姆萨染色和泛着丝粒FISH处理，以分析MN的组成，以区分生成素和破胚素的作用。Tukey多重比较检验的单因素方差分析显示(p <；0.001)，与未暴露的淋巴细胞培养相比，使用吉姆萨染色和着丝粒FISH双核细胞在所有这些基因毒性物质中观察到的MN增加。有着丝粒FISH的CBMN，在x射线、BLM（79.4%和79.2%）和MMC（81.5%和73.2%）下淋巴细胞中主要显示MNCN-ve细胞，而COL处理显著(p <；不同浓度的药物对MNCN+ve细胞的影响分别为79.6%和82.4%。着丝粒FISH能够鉴定不同基因毒性物质诱导的MN的来源。因此，在遗传毒理学的重大发展预示着在现代分析工作流程中纳入着丝粒探针。

{"title":"Use of centromeric probe to identify micronuclei origin and its advantages in genetic toxicology studies","authors":"G. Tamizh Selvan , P. Venkatachalam","doi":"10.1016/j.tiv.2025.106087","DOIUrl":"10.1016/j.tiv.2025.106087","url":null,"abstract":"<div><div>Fluorescence <em>in situ</em> hybridization (FISH) and karyotyping have long been considered essential for chromosomal examination to analyse the stable chromosome aberrations such as translocations after exposure to genotoxic agents. To improve speed of chromosomal analysis in genetic toxicology investigations, we employed the cytokinesis-block micronucleus (CBMN) assay with human pan-centromeric probes, to measure chromosome segregation errors and to differentiate mechanism of action of genotoxic agents. Whole-blood collected from healthy volunteers were exposed to X-rays, bleomycin (BLM), Colchicine (COL) and Mitomycin-C (MMC), arrested at cytokinesis stage and processed for conventional giemsa staining and pan-centromeric FISH to analyse the composition of MN to differentiate between aneugen and clastogen effects. One-way ANOVA with Tukey's multiple comparison test exhibited a significant (<em>p</em> < 0.001) increase in the MN observed using giemsa stained and centromere FISH binucleated cells in all those genotoxic agents when compared to its unexposed lymphocytes cultures. The CBMN with centromere FISH, predominantly shows MNCN<sup>-ve</sup> cells in lymphocytes exposed to X-rays (67.4 % and 83.3 %), BLM (79.4 % and 79.2 %) and MMC (81.5 % and 73.2) while, COL treatment resulted in significantly (<em>p</em> < 0.05) higher MNCN<sup>+ve</sup> cells (79.6 % and 82.4 %) for the different concentrations of those agents. Centromere FISH enables to identify the origin of MN induced by different genotoxic agents. Thus, a significant development in genetic toxicology is heralded by the incorporation of centromeric probe in modern analytical workflows.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"108 ","pages":"Article 106087"},"PeriodicalIF":2.6,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144168142","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}

引用次数: 0