Pub Date : 2025-09-25DOI: 10.1016/j.tiv.2025.106153
Dana Macejova , Jakub Kollar , Zdenek Dvorak , Daniela Schuster , Julius Brtko
Several organometallic trialkyltins or triaryltins are known to act as retinoid X receptor (RXR) agonists, which were also reported to exert eminent cytotoxic properties. In the present study, binding properties of tributyltin propionate (TBT-P) and tributyltin salicylate (TBT-S) in RXRα molecule were theoretically investigated using docking in Schrödinger small molecule drug discovery suite. Our data has shown that TBT-P and TBT-S bind in the binding pocket of RXRα and represent novel RXR agonists. In vitro data has shown that both TBT-P and TBT-S exert transcriptional activity under basal and ligand-activated conditions through RXR − nuclear thyroid hormone receptor (TR) heterodimer in the human reporter cell line for the assessment of thyroid receptor transcriptional activity (PZ-TR). In the human reporter cell line, enabling sensitive and selective identification of nuclear dihydroxyvitamin D3 receptor (VDR) agonists (IZ-VDRE), TBT-P and TBT-S did not exert any transcriptional activity through VDR under basal and ligand-activated conditions, which is in agreement with the properties of the non-permissive type of RXR-VDR heterodimer. Our in silico and in vitro data demonstrate that both organometallic compounds TBT-P and TBT-S represent new nuclear retinoid X receptor agonists.
{"title":"Tributyltin propionate and tributyltin salicylate represent novel RXR ligands: Effects on transcriptional activity of thyroid hormone receptor and vitamin D3 receptor","authors":"Dana Macejova , Jakub Kollar , Zdenek Dvorak , Daniela Schuster , Julius Brtko","doi":"10.1016/j.tiv.2025.106153","DOIUrl":"10.1016/j.tiv.2025.106153","url":null,"abstract":"<div><div>Several organometallic trialkyltins or triaryltins are known to act as retinoid X receptor (RXR) agonists, which were also reported to exert eminent cytotoxic properties. In the present study, binding properties of tributyltin propionate (TBT-P) and tributyltin salicylate (TBT-S) in RXRα molecule were theoretically investigated using docking in Schrödinger small molecule drug discovery suite. Our data has shown that TBT-P and TBT-S bind in the binding pocket of RXRα and represent novel RXR agonists. <em>In vitro</em> data has shown that both TBT-P and TBT-S exert transcriptional activity under basal and ligand-activated conditions through RXR − nuclear thyroid hormone receptor (TR) heterodimer in the human reporter cell line for the assessment of thyroid receptor transcriptional activity (PZ-TR). In the human reporter cell line, enabling sensitive and selective identification of nuclear dihydroxyvitamin D<sub>3</sub> receptor (VDR) agonists (IZ-VDRE), TBT-P and TBT-S did not exert any transcriptional activity through VDR under basal and ligand-activated conditions, which is in agreement with the properties of the non-permissive type of RXR-VDR heterodimer. Our <em>in silico</em> and <em>in vitro</em> data demonstrate that both organometallic compounds TBT-P and TBT-S represent new nuclear retinoid X receptor agonists.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106153"},"PeriodicalIF":2.7,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145158180","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}
Pub Date : 2025-09-24DOI: 10.1016/j.tiv.2025.106152
V. Fogaça-Santos , F.S. Alcântara , M.R.L. Conceição , L.P. Marques , J.L. Teixeira-Fonseca , D.J.B. Orts , J. Branquinho , R.L. Morais , K.O. Mota , D.S. Souza , J.B. Pesquero , D. Roman-Campos
Difenoconazole (DIF) is a widely used fungicide in agriculture, but it is unknown if it can modulate the excitability of the heart tissue. Thus, we investigated the acute effect of DIF on the electrical properties of heart tissue, with a particular focus on the cardiac sodium channel (Nav1.5) using transient expression of human Nav1.5 in human kidney cell (HEK) and human cardiomyocytes derived from induced pluripotent stem cells (CM-hiPSCs). DIF reduced the peak amplitude of sodium current (INa) in a concentration dependent manner. While DIF did not alter the voltage dependence of activation, it shifted the INa inactivation curve to more negative potentials and delayed the recovery from inactivation. In silico simulation identified aminoacids Thr1710 and Val1765 as critical for the interaction between DIF with Nav1.5. DIF partially blocked the late sodium current induced by deltamethrin, a pyrethroid pesticide, suggesting a potential interaction between these compounds in the environment. In CM-hiPSCs, DIF completely abolished the action potential. These findings suggest that DIF may modulate the biophysical properties of Nav1.5 and modulate cellular excitability which may cause reduced heart excitability, an DIF can also counteracting the effects of pyrethroid pesticides, but further studies using DIF concentrations closer to human exposure levels are necessary.
{"title":"The pesticide fungicide difenoconazole modulates the biophysical properties of sodium channel Nav1.5","authors":"V. Fogaça-Santos , F.S. Alcântara , M.R.L. Conceição , L.P. Marques , J.L. Teixeira-Fonseca , D.J.B. Orts , J. Branquinho , R.L. Morais , K.O. Mota , D.S. Souza , J.B. Pesquero , D. Roman-Campos","doi":"10.1016/j.tiv.2025.106152","DOIUrl":"10.1016/j.tiv.2025.106152","url":null,"abstract":"<div><div>Difenoconazole (DIF) is a widely used fungicide in agriculture, but it is unknown if it can modulate the excitability of the heart tissue. Thus, we investigated the acute effect of DIF on the electrical properties of heart tissue, with a particular focus on the cardiac sodium channel (Na<sub>v</sub>1.5) using transient expression of human Na<sub>v</sub>1.5 in human kidney cell (HEK) and human cardiomyocytes derived from induced pluripotent stem cells (CM-hiPSCs). DIF reduced the peak amplitude of sodium current (I<sub>Na</sub>) in a concentration dependent manner. While DIF did not alter the voltage dependence of activation, it shifted the I<sub>Na</sub> inactivation curve to more negative potentials and delayed the recovery from inactivation. <em>In silico</em> simulation identified aminoacids Thr1710 and Val1765 as critical for the interaction between DIF with Na<sub>v</sub>1.5. DIF partially blocked the late sodium current induced by deltamethrin, a pyrethroid pesticide, suggesting a potential interaction between these compounds in the environment. In CM-hiPSCs, DIF completely abolished the action potential. These findings suggest that DIF may modulate the biophysical properties of Na<sub>v</sub>1.5 and modulate cellular excitability which may cause reduced heart excitability, an DIF can also counteracting the effects of pyrethroid pesticides, but further studies using DIF concentrations closer to human exposure levels are necessary.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106152"},"PeriodicalIF":2.7,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145180280","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}
Pub Date : 2025-09-24DOI: 10.1016/j.tiv.2025.106149
Kamil Jurowski , Adrian Frydrych
This study provides the first toxicity profile of two aminoindane-based new psychoactive substances (NPS): 2-aminoindane (2-AI) and N-methyl-2-aminoindane (NM-2-AI), using a multi-method in silico approach. Tools including Percepta, ProTox 3.0, ADMETlab 3.0, VEGA QSAR, TEST, QSAR Toolbox, and STopTox were applied to evaluate acute toxicity, systemic organ effects, genotoxicity, skin and eye irritation, and cardiotoxicity. Both compounds were predicted to be acutely toxic via oral, dermal, and inhalation routes. Quantitative LD₅₀ values (rat, oral) ranged from 150 to 560 mg/kg (Percepta), 259.7–112.6 mg/kg (VEGA), and 326–360 mg/kg (ProTox 3.0). Health effect predictions indicated high probabilities of cardiovascular toxicity (2-AI: 80 %, NM-2-AI: 82 %) and pulmonary involvement (66 % and 65 %, respectively). NM-2-AI showed a higher likelihood of genotoxicity (Ames test) with a probability of 66.3 % (ADMETlab), compared to 2-AI (58.7 %). Both substances were predicted to be non-irritant to the eye, but skin irritation was probable for 2-AI (80 %, StopTox) and less so for NM-2-AI (60 %). hERG inhibition risk was low (IC₅₀: 391.7 μM for 2-AI; 126.9 μM for NM-2-AI). The in silico multi-tool strategy yielded convergent and interpretable data, supporting early toxicological characterization. These results are relevant for clinical management of NPS intoxications and for forensic interpretation in the absence of experimental data.
{"title":"First toxicity profile of aminoindane-based new psychoactive substances: 2-aminoindane (2-AI, CAS: 2975-41-9) and N-Methyl-2-aminoindane (NM-2-AI, CAS: 24445–44–1) – comprehensive prediction of toxicological endpoints important from clinical and forensic perspective using in silico multi- approach","authors":"Kamil Jurowski , Adrian Frydrych","doi":"10.1016/j.tiv.2025.106149","DOIUrl":"10.1016/j.tiv.2025.106149","url":null,"abstract":"<div><div>This study provides the first toxicity profile of two aminoindane-based new psychoactive substances (NPS): 2-aminoindane (2-AI) and <em>N</em>-methyl-2-aminoindane (NM-2-AI), using a multi-method <em>in silico</em> approach. Tools including Percepta, ProTox 3.0, ADMETlab 3.0, VEGA QSAR, TEST, QSAR Toolbox, and STopTox were applied to evaluate acute toxicity, systemic organ effects, genotoxicity, skin and eye irritation, and cardiotoxicity. Both compounds were predicted to be acutely toxic <em>via</em> oral, dermal, and inhalation routes. Quantitative LD₅₀ values (rat, oral) ranged from 150 to 560 mg/kg (Percepta), 259.7–112.6 mg/kg (VEGA), and 326–360 mg/kg (ProTox 3.0). Health effect predictions indicated high probabilities of cardiovascular toxicity (2-AI: 80 %, NM-2-AI: 82 %) and pulmonary involvement (66 % and 65 %, respectively). NM-2-AI showed a higher likelihood of genotoxicity (Ames test) with a probability of 66.3 % (ADMETlab), compared to 2-AI (58.7 %). Both substances were predicted to be non-irritant to the eye, but skin irritation was probable for 2-AI (80 %, StopTox) and less so for NM-2-AI (60 %). hERG inhibition risk was low (IC₅₀: 391.7 μM for 2-AI; 126.9 μM for NM-2-AI). The <em>in silico</em> multi-tool strategy yielded convergent and interpretable data, supporting early toxicological characterization. These results are relevant for clinical management of NPS intoxications and for forensic interpretation in the absence of experimental data.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106149"},"PeriodicalIF":2.7,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145180335","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}
Pub Date : 2025-09-20DOI: 10.1016/j.tiv.2025.106150
John F. Wambaugh , Nisha S. Sipes , Gilberto Padilla Mercado , Jon A. Arnot , Linda Bertato , Trevor N. Brown , Nicola Chirico , Christopher Cook , Daniel E. Dawson , Sarah E. Davidson-Fritz , Stephen S. Ferguson , Michael-Rock Goldsmith , Chris M. Grulke , Richard S. Judson , Kamel Mansouri , Grace Patlewicz , Ester Papa , Prachi Pradeep , Alessandro Sangion , Risa R. Sayre , Michael J. Devito
High throughput toxicokinetic (HTTK) methods address chemical risk assessment data gaps but require chemical-specific values that can be obtained by in vitro measurements or in silico models. In this study, seven quantitative structure property relationship (QSPR) models were used to estimate intrinsic hepatic clearance (Clint), fraction of chemical unbound in plasma (fup), and/or TK elimination half-life (t½). Performance of the QSPR models was evaluated using literature time-course in vivo TK data, mainly from rats. Simulations of the in vivo data were made with a high throughput physiologically based TK (HT-PBTK) model using the different QSPR model predictions as inputs. We estimate that using rat in vivo data to evaluate QSPR models trained on human in vitro measured data might inflate error estimates by as much as root mean squared log10 error (RMSLE) 0.8. A sensitivity analysis showed that Clint and fup parameters inform predictions of area under the curve (AUC) and steady-state concentration (Css). We estimate that AUC can be predicted by HTTK with RMSLE 0.9 using in vitro measurements and 0.6–0.8 using QSPR model values. We anticipate that, for some novel compounds, QSPRs for HTTK input parameters will give predictions of TK similar to those based on in vitro measurements.
{"title":"Collaborative evaluation of in silico predictions for high throughput toxicokinetics","authors":"John F. Wambaugh , Nisha S. Sipes , Gilberto Padilla Mercado , Jon A. Arnot , Linda Bertato , Trevor N. Brown , Nicola Chirico , Christopher Cook , Daniel E. Dawson , Sarah E. Davidson-Fritz , Stephen S. Ferguson , Michael-Rock Goldsmith , Chris M. Grulke , Richard S. Judson , Kamel Mansouri , Grace Patlewicz , Ester Papa , Prachi Pradeep , Alessandro Sangion , Risa R. Sayre , Michael J. Devito","doi":"10.1016/j.tiv.2025.106150","DOIUrl":"10.1016/j.tiv.2025.106150","url":null,"abstract":"<div><div>High throughput toxicokinetic (HTTK) methods address chemical risk assessment data gaps but require chemical-specific values that can be obtained by <em>in vitro</em> measurements or <em>in silico</em> models. In this study, seven quantitative structure property relationship (QSPR) models were used to estimate intrinsic hepatic clearance (Cl<sub>int</sub>), fraction of chemical unbound in plasma (f<sub>up</sub>), and/or TK elimination half-life (t<sub>½</sub>). Performance of the QSPR models was evaluated using literature time-course <em>in vivo</em> TK data, mainly from rats. Simulations of the <em>in vivo</em> data were made with a high throughput physiologically based TK (HT-PBTK) model using the different QSPR model predictions as inputs. We estimate that using rat <em>in vivo</em> data to evaluate QSPR models trained on human <em>in vitro</em> measured data might inflate error estimates by as much as root mean squared log<sub>10</sub> error (RMSLE) 0.8. A sensitivity analysis showed that Cl<sub>int</sub> and f<sub>up</sub> parameters inform predictions of area under the curve (AUC) and steady-state concentration (C<sub>ss</sub>). We estimate that AUC can be predicted by HTTK with RMSLE 0.9 using <em>in vitro</em> measurements and 0.6–0.8 using QSPR model values. We anticipate that, for some novel compounds, QSPRs for HTTK input parameters will give predictions of TK similar to those based on <em>in vitro</em> measurements.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106150"},"PeriodicalIF":2.7,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145126470","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}
Our study explores the effects of glucose, fructose, and sucrose on hepatocyte health, focusing on cell viability and lipid accumulation. Hepatocyte cells (HepG2) were exposed to a range of concentrations of glucose, fructose and sucrose, both individually and in combination, over 24, 48, and 72 h. Cell viability was monitored using the MTT assay, while lipid accumulation was estimated fluorometrically using Nile red staining. Our findings suggest that higher concentrations of fructose significantly improves cell viability in a dose-dependent manner, especially in hepatocyte cells exposed to fructose for extended duration. Additionally, a time-dependent increase in lipid accumulation was seen, particularly in cells exposed to fructose. Interestingly, enhanced hepatocyte viability was noted with combinations of glucose with fructose or sucrose, without detrimental effects. Lipid accumulation was more pronounced with glucose-fructose combinations. These results provide insights into the differential effects of dietary sugars on liver health and metabolism, with implications for understanding metabolic disorders like MASLD and MASH.
{"title":"Synergistic and divergent effects of dietary sugars on hepatocyte viability and steatosis","authors":"Ajay Kumar , Garhima Arora , Deepika Kumari , Harish Changotra , Samrat Chatterjee","doi":"10.1016/j.tiv.2025.106148","DOIUrl":"10.1016/j.tiv.2025.106148","url":null,"abstract":"<div><div>Our study explores the effects of glucose, fructose, and sucrose on hepatocyte health, focusing on cell viability and lipid accumulation. Hepatocyte cells (HepG2) were exposed to a range of concentrations of glucose, fructose and sucrose, both individually and in combination, over 24, 48, and 72 h. Cell viability was monitored using the MTT assay, while lipid accumulation was estimated fluorometrically using Nile red staining. Our findings suggest that higher concentrations of fructose significantly improves cell viability in a dose-dependent manner, especially in hepatocyte cells exposed to fructose for extended duration. Additionally, a time-dependent increase in lipid accumulation was seen, particularly in cells exposed to fructose. Interestingly, enhanced hepatocyte viability was noted with combinations of glucose with fructose or sucrose, without detrimental effects. Lipid accumulation was more pronounced with glucose-fructose combinations. These results provide insights into the differential effects of dietary sugars on liver health and metabolism, with implications for understanding metabolic disorders like MASLD and MASH.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106148"},"PeriodicalIF":2.7,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145103243","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}
Pub Date : 2025-09-18DOI: 10.1016/j.tiv.2025.106146
G. Drouet , A. Bourgois , K. Devilliers , M. Defrance , A. Van der Meeren
Pulmonary absorption is a critical step in the time-dependent processes governing actinide biodistribution and toxicity following internal contamination by inhalation. The current study was undertaken to investigate on a simple in vitro Calu-3 model the mechanisms involved in the transfer across epithelial cells of plutonium (Pu) and americium (Am) as a function of their intrinsic properties (nature, physicochemical form) or their association with serum proteins, inorganic ligands or synthetic chelating agents. Both the contribution of the transcellular and paracellular routes to the transfer of these various forms were studied. Results show that uptake of Pu and Am by epithelial cells is not positively correlated with their transfer, which suggests that radionuclides transcytosis is a continuous process in comparison with the quick translocation of Pu/Am-small size complexes. Furthermore, reducing epithelial cells paracellular permeability with the glucocorticoid hormone Dexamethasone (DEX) resulted into a lower transfer of Pu citrate, and of both actinides in the presence of diethylene triamine pentaacetic acid (DTPA) and 3,4,3-LI(1,2-HOPO). Taken together, these results seem to indicate that a paracellular pathway may be responsible, at least partly, for the higher pulmonary absorption observed in vivo for actinides bound to low-molecular weight ligands such as citrate, DTPA or HOPO.
肺吸收是控制锕系元素生物分布和吸入内部污染后毒性的时间依赖过程中的关键步骤。目前的研究是在一个简单的体外Calu-3模型上进行的，以研究钚（Pu）和镅（Am）的内在特性（性质、物理化学形式）或它们与血清蛋白、无机配体或合成螯合剂的关联在上皮细胞间转移的机制。研究了跨细胞和细胞旁途径对这些不同形式的转移的贡献。结果表明，上皮细胞对Pu和Am的摄取与它们的转移不呈正相关，这表明与Pu/Am小尺寸复合物的快速易位相比，放射性核素的胞吞作用是一个连续的过程。此外，糖皮质激素地塞米松（Dexamethasone， DEX）降低上皮细胞的细胞旁通透性，导致在二乙烯三胺五乙酸（DTPA）和3,4,3- li （1,2- hopo）存在下，柠檬酸Pu和两种锕系元素的转移降低。综上所述，这些结果似乎表明，细胞旁通路可能至少在一定程度上负责观察到锕系元素与低分子量配体（如柠檬酸盐、DTPA或HOPO）结合的较高肺吸收。
{"title":"Binding of plutonium and americium to small molecular weight ligands favors their paracellular transfer across lung epithelium in vitro","authors":"G. Drouet , A. Bourgois , K. Devilliers , M. Defrance , A. Van der Meeren","doi":"10.1016/j.tiv.2025.106146","DOIUrl":"10.1016/j.tiv.2025.106146","url":null,"abstract":"<div><div>Pulmonary absorption is a critical step in the time-dependent processes governing actinide biodistribution and toxicity following internal contamination by inhalation. The current study was undertaken to investigate on a simple in vitro Calu-3 model the mechanisms involved in the transfer across epithelial cells of plutonium (Pu) and americium (Am) as a function of their intrinsic properties (nature, physicochemical form) or their association with serum proteins, inorganic ligands or synthetic chelating agents. Both the contribution of the transcellular and paracellular routes to the transfer of these various forms were studied. Results show that uptake of Pu and Am by epithelial cells is not positively correlated with their transfer, which suggests that radionuclides transcytosis is a continuous process in comparison with the quick translocation of Pu/Am-small size complexes. Furthermore, reducing epithelial cells paracellular permeability with the glucocorticoid hormone Dexamethasone (DEX) resulted into a lower transfer of Pu citrate, and of both actinides in the presence of diethylene triamine pentaacetic acid (DTPA) and 3,4,3-LI(1,2-HOPO). Taken together, these results seem to indicate that a paracellular pathway may be responsible, at least partly, for the higher pulmonary absorption observed in vivo for actinides bound to low-molecular weight ligands such as citrate, DTPA or HOPO.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106146"},"PeriodicalIF":2.7,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145103256","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}
Pub Date : 2025-09-17DOI: 10.1016/j.tiv.2025.106147
Saja Damdam , Blake Hunnie , Parker Jones , Markus Hecker , Markus Brinkmann , Francisco C. da Silva Junior
N-(1,3-Dimethylbutyl)-N′-phenyl-p-phenylenediamine-quinone (6PPD-Q), an environmental transformation product of the rubber tire antioxidant 6PPD, is increasingly detected in urban environments, yet its impact on human health. This study investigated toxicological effects of 6PPD-Q in two human lung cells: BEAS-2B (a transformed bronchial epithelial cell) and A549 (a lung adenocarcinoma cell). Cells were exposed to 6PPD-Q at concentrations of 5 to 80 ng/mL and effects on cell viability, oxidative stress by generation of reactive oxygen species (ROS), disruption of antioxidant defense response, oxidative DNA damage as determined by formation of 8-hydroxy-2′-deoxyguanosine (8-OHdG), and biotransformation 6PPD-Q were determined. Results showed a biphasic response with an increase in cell viability at lower concentrations, suggesting an adaptive cellular response. ROS production and levels of 8-OHdG significantly increased, indicating an imbalance in redox status and induction of oxidative DNA damage. Two phase I hydroxylation products, 6PPD-Q-4-OH (TPOH1) and 6PPD-Q-phenyl-OH (TPOH2), were identified. In A549 cells, TPOH1 was only in the media, while TPOH2 was found both inside cells and in the media. Moreover, mRNA expression analysis revealed upregulation of metabolism- and oxidative stress-related genes and inflammatory cytokines. These findings suggest that 6PPD-Q may trigger oxidative stress and inflammation in lung cells. This study highlights the effects of 6PPD-Q on lung cells and underscores the need for further research on its long-term impact on human respiratory health.
{"title":"Toxicity assessment of 6PPD-quinone in human lung cells: Insights from BEAS-2B and A549 cell lines","authors":"Saja Damdam , Blake Hunnie , Parker Jones , Markus Hecker , Markus Brinkmann , Francisco C. da Silva Junior","doi":"10.1016/j.tiv.2025.106147","DOIUrl":"10.1016/j.tiv.2025.106147","url":null,"abstract":"<div><div>N-(1,3-Dimethylbutyl)-<em>N</em>′-phenyl-<em>p</em>-phenylenediamine-quinone (6PPD-Q), an environmental transformation product of the rubber tire antioxidant 6PPD, is increasingly detected in urban environments, yet its impact on human health. This study investigated toxicological effects of 6PPD-Q in two human lung cells: BEAS-2B (a transformed bronchial epithelial cell) and A549 (a lung adenocarcinoma cell). Cells were exposed to 6PPD-Q at concentrations of 5 to 80 ng/mL and effects on cell viability, oxidative stress by generation of reactive oxygen species (ROS), disruption of antioxidant defense response, oxidative DNA damage as determined by formation of 8-hydroxy-2′-deoxyguanosine (8-OHdG), and biotransformation 6PPD-Q were determined. Results showed a biphasic response with an increase in cell viability at lower concentrations, suggesting an adaptive cellular response. ROS production and levels of 8-OHdG significantly increased, indicating an imbalance in redox status and induction of oxidative DNA damage. Two phase I hydroxylation products, 6PPD-Q-4-OH (TPOH1) and 6PPD-Q-phenyl-OH (TPOH2), were identified. In A549 cells, TPOH1 was only in the media, while TPOH2 was found both inside cells and in the media. Moreover, mRNA expression analysis revealed upregulation of metabolism- and oxidative stress-related genes and inflammatory cytokines. These findings suggest that 6PPD-Q may trigger oxidative stress and inflammation in lung cells. This study highlights the effects of 6PPD-Q on lung cells and underscores the need for further research on its long-term impact on human respiratory health.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106147"},"PeriodicalIF":2.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145092993","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}
Pub Date : 2025-09-16DOI: 10.1016/j.tiv.2025.106127
Veronica M. Lucian, Robert C. Carlisle, Constantin C. Coussios, Malavika Nair
Testing cell viability in three-dimensional (3D) in vitro cell cultures is not always straightforward. Reduction-based metabolic assays have been suggested as a rapid, reportedly non-toxic mechanism for monitoring cell viability in continuous culture experiments. However, in 3D cultures these assays have been predominantly used semiquantitatively to indicate relative trends in viability. Obtaining quantitative values of cell number is often challenging, particularly when performing continuous measurements of viability when compared with end-point assessments. This work presents experiments performed to robustly identify and address the sources of inconsistency in existing Presto Blue™ methods for 3D cultures. Using human dermal fibroblasts (HDFs) as a model cell line seeded onto type I collagen scaffolds, we examine the impact of Presto Blue™ assays on reported cell numbers in 2D versus 3D cultures. We separate the impact of potential dye-induced toxicity from manipulation-induced cell detachment by considering the effects of passage number, incubation period, modifications of substrate chemistry and duration of total dye exposure as independent variables. The work presented includes a finalised protocol that demonstrates that while resazurin-based reduction assays remain a valuable tool for rapid assessment of cell metabolism and/or viability, additional considerations of the substrate and sampling frequencies are needed in 3D cultures to ensure accurate and reproducible results.
{"title":"Use of resazurin-based metabolic assays as a continuous measure of viability in three-dimensional (3D) in vitro cell culture: A study with Presto Blue™","authors":"Veronica M. Lucian, Robert C. Carlisle, Constantin C. Coussios, Malavika Nair","doi":"10.1016/j.tiv.2025.106127","DOIUrl":"10.1016/j.tiv.2025.106127","url":null,"abstract":"<div><div>Testing cell viability in three-dimensional (3D) <em>in vitro</em> cell cultures is not always straightforward. Reduction-based metabolic assays have been suggested as a rapid, reportedly non-toxic mechanism for monitoring cell viability in continuous culture experiments. However, in 3D cultures these assays have been predominantly used semiquantitatively to indicate relative trends in viability. Obtaining quantitative values of cell number is often challenging, particularly when performing continuous measurements of viability when compared with end-point assessments. This work presents experiments performed to robustly identify and address the sources of inconsistency in existing Presto Blue™ methods for 3D cultures. Using human dermal fibroblasts (HDFs) as a model cell line seeded onto type I collagen scaffolds, we examine the impact of Presto Blue™ assays on reported cell numbers in 2D versus 3D cultures. We separate the impact of potential dye-induced toxicity from manipulation-induced cell detachment by considering the effects of passage number, incubation period, modifications of substrate chemistry and duration of total dye exposure as independent variables. The work presented includes a finalised protocol that demonstrates that while resazurin-based reduction assays remain a valuable tool for rapid assessment of cell metabolism and/or viability, additional considerations of the substrate and sampling frequencies are needed in 3D cultures to ensure accurate and reproducible results.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106127"},"PeriodicalIF":2.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145088106","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}
Pub Date : 2025-09-05DOI: 10.1016/j.tiv.2025.106145
K. Bobadilla-Lozoya , J. Morales-Montor , K.G. Mejía-Salgado , K.E. Nava-Castro
Human activity has led to the increment of diverse pollutants. Plastics have great practical value since they are present in everyday products. However, not only plastics have gained importance, but their plasticizers such as bisphenol A (BPA), phthalates and other chemicals such as the polyaromatic hydrocarbon compounds (PAHs) have described to impact in human and animal health because of its chronic exposure and that they are endocrine disruptors (EDs). Since immune cells express hormone receptors and AhR, they are susceptible to these EDs. We hypothesized that these compounds may alter their maturation and function. We focus on macrophages since they link the innate and adaptive immune response, playing different roles in disease. We cultured human monocytes in the presence of phthalates, bisphenol A or PAH's and analyze both maturation and function such as phagocytosis, antigen processing and antigen presentation. Our results show that these compounds affect differentially macrophage function, being the phtalates DEHP and DBP the ones that have the greatest impact. Both inhibit human macrophage maturation, observed by the lack of expression of MHC-II, CD80/86, which in consequence, impair antigen presentation and phagocytosis. BPA did not alter maturation but changed antigen processing activity while PAHs alter both antigen processing and presentation. These results may have great implications on human health and disease, since emergent pollutants affect normal macrophage function.
{"title":"The emerging pollutants polycyclic aromatic compounds (PAH'S) bisphenol a (BPA), and phthalates impair immune system function: Effects on human macrophages","authors":"K. Bobadilla-Lozoya , J. Morales-Montor , K.G. Mejía-Salgado , K.E. Nava-Castro","doi":"10.1016/j.tiv.2025.106145","DOIUrl":"10.1016/j.tiv.2025.106145","url":null,"abstract":"<div><div>Human activity has led to the increment of diverse pollutants. Plastics have great practical value since they are present in everyday products. However, not only plastics have gained importance, but their plasticizers such as bisphenol A (BPA), phthalates and other chemicals such as the polyaromatic hydrocarbon compounds (PAHs) have described to impact in human and animal health because of its chronic exposure and that they are endocrine disruptors (EDs). Since immune cells express hormone receptors and AhR, they are susceptible to these EDs. We hypothesized that these compounds may alter their maturation and function. We focus on macrophages since they link the innate and adaptive immune response, playing different roles in disease. We cultured human monocytes in the presence of phthalates, bisphenol A or PAH's and analyze both maturation and function such as phagocytosis, antigen processing and antigen presentation. Our results show that these compounds affect differentially macrophage function, being the phtalates DEHP and DBP the ones that have the greatest impact. Both inhibit human macrophage maturation, observed by the lack of expression of MHC-II, CD80/86, which in consequence, impair antigen presentation and phagocytosis. BPA did not alter maturation but changed antigen processing activity while PAHs alter both antigen processing and presentation. These results may have great implications on human health and disease, since emergent pollutants affect normal macrophage function.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106145"},"PeriodicalIF":2.7,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145014410","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}
Pub Date : 2025-09-03DOI: 10.1016/j.tiv.2025.106144
Nikita N. Chukavin , Anton L. Popov , Victoria A. Anikina , Daria A. Vinnik , Boris A. Bokl , Nelli R. Popova
Curcumin is a natural bioactive substance with promising biomedical applications. However, the low solubility and stability of curcumin significantly limit its potential use. The development of nanoformulations of curcumin makes it possible to circumvent the above limitations. Cerium dioxide (CeO2, NDC) nanoparticles are a promising platform for curcumin binding. They are able to absorb curcumin on their surface, providing increased bioavailability and bioaccumulation. Moreover, NDC have unique enzyme-like properties that can be used for targeted delivery and controlled release of curcumin. Meanwhile, the potential cytotoxicity of such nanoformulations remains poorly understood. In this work, we synthesized the NDC-curcumin nanoconjugate and investigated the effect of excess curcumin on the cytotoxicity of this nanoformulation. Curcumin has been shown to bind to the surface of nanoparticles, forming a colloidally stable nanoconjugate. At the same time, excess curcumin formed a separate nanoscale fraction, which caused the statistically significant cytotoxicity of the nanoconjugate in relation to human keratinocytes (HaCaT), mesenchymal stem cells (hMSc) and fibroblasts (HF). These cell lines represent the cells that form skin, mucous membranes, and connective tissue, with which curcumin nanoformulations can interact through various methods of administration. IC50 values of curcumin and NDC-curcumin were 400 and 482 μM for HaCaT, 193 and 211 μM for hMSc, 266 and 304 μM for HF, respectively, after 72 h of coincubation. Consequently, NDC provided an increased biocompatibility of curcumin. Application of a rational design of curcumin nanoformulations can help to overcome possible limitations of its practical use due to its toxicity, enhancing its bioactivity.
{"title":"Excess curcumin causes cytotoxicity of its nanoconjugate with nanoceria","authors":"Nikita N. Chukavin , Anton L. Popov , Victoria A. Anikina , Daria A. Vinnik , Boris A. Bokl , Nelli R. Popova","doi":"10.1016/j.tiv.2025.106144","DOIUrl":"10.1016/j.tiv.2025.106144","url":null,"abstract":"<div><div>Curcumin is a natural bioactive substance with promising biomedical applications. However, the low solubility and stability of curcumin significantly limit its potential use. The development of nanoformulations of curcumin makes it possible to circumvent the above limitations. Cerium dioxide (CeO<sub>2</sub>, NDC) nanoparticles are a promising platform for curcumin binding. They are able to absorb curcumin on their surface, providing increased bioavailability and bioaccumulation. Moreover, NDC have unique enzyme-like properties that can be used for targeted delivery and controlled release of curcumin. Meanwhile, the potential cytotoxicity of such nanoformulations remains poorly understood. In this work, we synthesized the NDC-curcumin nanoconjugate and investigated the effect of excess curcumin on the cytotoxicity of this nanoformulation. Curcumin has been shown to bind to the surface of nanoparticles, forming a colloidally stable nanoconjugate. At the same time, excess curcumin formed a separate nanoscale fraction, which caused the statistically significant cytotoxicity of the nanoconjugate in relation to human keratinocytes (HaCaT), mesenchymal stem cells (hMSc) and fibroblasts (HF). These cell lines represent the cells that form skin, mucous membranes, and connective tissue, with which curcumin nanoformulations can interact through various methods of administration. IC<sub>50</sub> values of curcumin and NDC-curcumin were 400 and 482 μM for HaCaT, 193 and 211 μM for hMSc, 266 and 304 μM for HF, respectively, after 72 h of coincubation. Consequently, NDC provided an increased biocompatibility of curcumin. Application of a rational design of curcumin nanoformulations can help to overcome possible limitations of its practical use due to its toxicity, enhancing its bioactivity.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"110 ","pages":"Article 106144"},"PeriodicalIF":2.7,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145004682","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}
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