Extended oral intake of silver nanoparticles (AgNP) may result in significant exposure of intestinal cells, which could trigger intestinal inflammation, an effect that requires deep comprehension and the development of mitigation measures. This study aimed to investigate the potential pro-inflammatory effects of polyvinylpyrrolidone (PVP)-coated AgNP with two different sizes (5 and 50 nm) on human intestinal epithelial C2BBe1 cells, as well as the potential mitigation effects of quercetin, a flavonoid with acknowledged antioxidant and anti-inflammatory potential. The pro-inflammatory effects of PVP-AgNP on several proteins related to the intestinal inflammatory response, as well as on ●NO production and cytokine production, were investigated. PVP-AgNP exposure caused cellular stress and damage, as evidenced by a reduction in cellular metabolic activity, ultrastructural changes, and loss of viability. The present study also found that 5 nm PVP-AgNP triggered inflammation through the p65-induced NF-κB pathway and reduced p-Nrf2 expression, while 50 nm PVP-AgNP activated the IκBα-induced NF-κB pathway and elevated COX-2 levels. Both sizes induced an increase of PGE2 levels and IL-8 production, with only 5 nm PVP-AgNP increasing IL-6. The harmful effects of PVP-AgNP were effectively mitigated by quercetin, which highlights the potential of this flavonoid to modulate the respective potential damage at the intestinal level.
{"title":"Quercetin mitigates pro-inflammatory effects of polyvinylpyrrolidone-coated silver nanoparticles on human intestinal epithelial cells","authors":"Adelaide Sousa , Inês Santos , Rui Fernandes , Sofia Pacheco , Félix Carvalho , Eduarda Fernandes , Marisa Freitas","doi":"10.1016/j.tiv.2025.106176","DOIUrl":"10.1016/j.tiv.2025.106176","url":null,"abstract":"<div><div>Extended oral intake of silver nanoparticles (AgNP) may result in significant exposure of intestinal cells, which could trigger intestinal inflammation, an effect that requires deep comprehension and the development of mitigation measures. This study aimed to investigate the potential pro-inflammatory effects of polyvinylpyrrolidone (PVP)-coated AgNP with two different sizes (5 and 50 nm) on human intestinal epithelial C2BBe1 cells, as well as the potential mitigation effects of quercetin, a flavonoid with acknowledged antioxidant and anti-inflammatory potential. The pro-inflammatory effects of PVP-AgNP on several proteins related to the intestinal inflammatory response, as well as on <sup>●</sup>NO production and cytokine production, were investigated. PVP-AgNP exposure caused cellular stress and damage, as evidenced by a reduction in cellular metabolic activity, ultrastructural changes, and loss of viability. The present study also found that 5 nm PVP-AgNP triggered inflammation through the p65-induced NF-κB pathway and reduced p-Nrf2 expression, while 50 nm PVP-AgNP activated the IκBα-induced NF-κB pathway and elevated COX-2 levels. Both sizes induced an increase of PGE<sub>2</sub> levels and IL-8 production, with only 5 nm PVP-AgNP increasing IL-6. The harmful effects of PVP-AgNP were effectively mitigated by quercetin, which highlights the potential of this flavonoid to modulate the respective potential damage at the intestinal level.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"112 ","pages":"Article 106176"},"PeriodicalIF":2.7,"publicationDate":"2025-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145598126","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 : 2025-11-21DOI: 10.1016/j.tiv.2025.106177
James W. Simpkins, Ralph L. Cooper, Charles Breckenridge
This study investigated the effects of atrazine (ATZ) and its metabolites on phosphodiesterase (PDE) activity ex vivo and aromatase mRNA expression in two steroidogenic cell lines (H295R and JEG-3). The pharmacokinetics of ATZ and its chlorometabolites were characterized for intact female Sprague-Dawley (SD) rats. A “Quantitative In Vitro to In Vivo Extrapolation” (QIVIVE) method was developed and used to calculate the margins of exposure to chlorotriazines in humans. Atrazine, and to a much lesser extent, its metabolites, resulted in PDE inhibition ex vivo. The IC50 for atrazine was 39.52 μM. Neither H295R nor JEG-3 cells metabolize atrazine in vitro. 10 μM atrazine induced a 2- to 3-fold increase in aromatase mRNA expression in H295R cells. JEG3 cells gave similar results.
Pharmacokinetic parameters for the chlorotriazines in female SD rats indicated that atrazine is rapidly absorbed and cleared from plasma after a bolus dose. QIVIVE analyses revealed the AUC at the No Observed Effect Concentration (NOEC) for atrazine inhibition of PDE ex vivo following 1 h of exposure was 350- to 1013-fold greater than the AUC plasma concentration following gavage or dietary dosing in vivo at the lowest NOEL currently used as a point of departure (POD) in human risk assessment. These results indicate that atrazine-induced PDE inhibition in vitro would not have a functional effect in vivo at the POD from animal studies or in humans exposed to atrazine via diet, drinking water or occupationally.
{"title":"Quantitative in vitro to in vivo extrapolation (QIVIVE) of atrazine's effects on phosphodiesterase and aromatase mRNA expression in vitro to the rat in vivo based on pharmacokinetic","authors":"James W. Simpkins, Ralph L. Cooper, Charles Breckenridge","doi":"10.1016/j.tiv.2025.106177","DOIUrl":"10.1016/j.tiv.2025.106177","url":null,"abstract":"<div><div>This study investigated the effects of atrazine (ATZ) and its metabolites on phosphodiesterase (PDE) activity <em>ex vivo</em> and aromatase mRNA expression in two steroidogenic cell lines (H295R and JEG-3). The pharmacokinetics of ATZ and its chlorometabolites were characterized for intact female Sprague-Dawley (SD) rats. A “Quantitative <em>In Vitro</em> to <em>In Vivo</em> Extrapolation” (QIVIVE) method was developed and used to calculate the margins of exposure to chlorotriazines in humans. Atrazine, and to a much lesser extent, its metabolites, resulted in PDE inhibition <em>ex vivo</em>. The IC<sub>50</sub> for atrazine was 39.52 μM. Neither H295R nor JEG-3 cells metabolize atrazine <em>in vitro</em>. 10 μM atrazine induced a 2- to 3-fold increase in aromatase mRNA expression in H295R cells. JEG3 cells gave similar results.</div><div>Pharmacokinetic parameters for the chlorotriazines in female SD rats indicated that atrazine is rapidly absorbed and cleared from plasma after a bolus dose. QIVIVE analyses revealed the AUC at the No Observed Effect Concentration (NOEC) for atrazine inhibition of PDE <em>ex vivo</em> following 1 h of exposure was 350- to 1013-fold greater than the AUC plasma concentration following gavage or dietary dosing <em>in vivo</em> at the lowest NOEL currently used as a point of departure (POD) in human risk assessment. These results indicate that atrazine-induced PDE inhibition <em>in vitro</em> would not have a functional effect <em>in vivo</em> at the POD from animal studies or in humans exposed to atrazine <em>via</em> diet, drinking water or occupationally.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106177"},"PeriodicalIF":2.7,"publicationDate":"2025-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589828","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 : 2025-11-20DOI: 10.1016/j.tiv.2025.106178
Jakeline Liara Teleken , Leticia Prates Roma , Rosane Aparecida Ribeiro , Alex Rafacho , Maria Lúcia Bonfleur
Glyphosate (GLY) is among the most widely used pesticides globally. Emerging evidence suggests that GLY-based herbicides (GBHs) are more toxic than GLY alone. In this study, we used HepG2 cells expressing mitochondrial-targeted Hyper7 to monitor H₂O₂ production and performed RNA sequencing to compare transcriptomic responses to GLY and a commercial GBH formulation. GBH exposure significantly elevated mitochondrial H₂O₂ levels compared to control and GLY-treated cells. Transcriptomic analysis revealed upregulation of gene ontology (GO) terms associated with oxidative stress and response to hydrogen peroxide, alongside downregulation of antioxidant enzyme genes and reduced PRDX3 protein, indicating impaired redox homeostasis in GBH-exposed cells. GBH also induced ER stress, marked by increased expression of stress-related genes (Ern1, Ddit3) and enrichment of GO terms for the unfolded protein response. GBH treatment upregulated genes involved in autophagy and apoptosis (Sqstm1, Bbc3, Map1lc3b), suggesting progressive stress response. Additionally, in GBH-treated cells metabolic pathways were altered, with higher expression of Atf3 and Ppargc1a, and enrichment of GO terms including response to glucose and lipid metabolism regulation These results indicate that GBH triggers complex stress and metabolic changes distinct from GLY alone, highlighting the importance of assessing full formulations to better evaluate potential health risks.
{"title":"Commercial glyphosate formulations exceed active ingredient toxicity via mitochondrial ROS and transcriptomic disruption","authors":"Jakeline Liara Teleken , Leticia Prates Roma , Rosane Aparecida Ribeiro , Alex Rafacho , Maria Lúcia Bonfleur","doi":"10.1016/j.tiv.2025.106178","DOIUrl":"10.1016/j.tiv.2025.106178","url":null,"abstract":"<div><div>Glyphosate (GLY) is among the most widely used pesticides globally. Emerging evidence suggests that GLY-based herbicides (GBHs) are more toxic than GLY alone. In this study, we used HepG2 cells expressing mitochondrial-targeted Hyper7 to monitor H₂O₂ production and performed RNA sequencing to compare transcriptomic responses to GLY and a commercial GBH formulation. GBH exposure significantly elevated mitochondrial H₂O₂ levels compared to control and GLY-treated cells. Transcriptomic analysis revealed upregulation of gene ontology (GO) terms associated with oxidative stress and response to hydrogen peroxide, alongside downregulation of antioxidant enzyme genes and reduced PRDX3 protein, indicating impaired redox homeostasis in GBH-exposed cells. GBH also induced ER stress, marked by increased expression of stress-related genes (Ern1, Ddit3) and enrichment of GO terms for the unfolded protein response. GBH treatment upregulated genes involved in autophagy and apoptosis (Sqstm1, Bbc3, Map1lc3b), suggesting progressive stress response. Additionally, in GBH-treated cells metabolic pathways were altered, with higher expression of Atf3 and Ppargc1a, and enrichment of GO terms including response to glucose and lipid metabolism regulation These results indicate that GBH triggers complex stress and metabolic changes distinct from GLY alone, highlighting the importance of assessing full formulations to better evaluate potential health risks.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106178"},"PeriodicalIF":2.7,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145582816","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 : 2025-11-17DOI: 10.1016/j.tiv.2025.106175
Ji-Woo Choe , Geon-Hee Lee , Su-Hyun Lee , Ga-Eun Kim , Ha-Ryong Kim , Kyung-Min Lim
This study presents a novel in vitro method for acute respiratory toxicity testing using the SoluAirway™ model (SoluAirway™ ARTT). SoluAirway™ is a 3D human airway tissue model constructed from primary human nasal epithelial cells, recapitulating the structural and molecular characteristics of the human bronchial epithelium. Tissues were exposed to four concentrations of test chemicals using two application methods—vapor-cap and direct application—for 4 h, followed by a 20 h post-incubation. Tissue viability was assessed using the MTT assay. Following established classification approaches, EC₅₀ and EC₂₅ values were determined and used to assign GHS acute inhalation toxicity categories: EC₂₅/₅₀ ≤ 5 mg/tissue: GHS Category 1 or 2, 5 < EC₂₅/₅₀ ≤ 25 mg/tissue: GHS Category 3 or 4 and EC₂₅/₅₀ > 25 mg/tissue: No Category. The direct application method showed an excellent predictive accuracy of 76.92 % (10/13). When 20 additional reference chemicals (33 total) were tested, the EC₅₀-based classification achieved a predictive accuracy of 75.76 % (25/33). Notably, applying a fixed-dose viability threshold (≤50 % viability at 5 mg/tissue for GHS 1/2; >50 % at 5 and ≤ 50 % at 25 mg/tissue for GHS 3/4; others as No Category) yielded comparable predictive performance. Collectively, these results suggest that SoluAirway™ ARTT is a promising, cost-effective alternative method for assessing acute inhalation toxicity.
{"title":"Development of an in vitro acute respiratory toxicity test using the SoluAirway™, 3D human airway model","authors":"Ji-Woo Choe , Geon-Hee Lee , Su-Hyun Lee , Ga-Eun Kim , Ha-Ryong Kim , Kyung-Min Lim","doi":"10.1016/j.tiv.2025.106175","DOIUrl":"10.1016/j.tiv.2025.106175","url":null,"abstract":"<div><div>This study presents a novel in vitro method for acute respiratory toxicity testing using the SoluAirway™ model (SoluAirway™ ARTT). SoluAirway™ is a 3D human airway tissue model constructed from primary human nasal epithelial cells, recapitulating the structural and molecular characteristics of the human bronchial epithelium. Tissues were exposed to four concentrations of test chemicals using two application methods—vapor-cap and direct application—for 4 h, followed by a 20 h post-incubation. Tissue viability was assessed using the MTT assay. Following established classification approaches, EC₅₀ and EC₂₅ values were determined and used to assign GHS acute inhalation toxicity categories: EC₂₅/₅₀ ≤ 5 mg/tissue: GHS Category 1 or 2, 5 < EC₂₅/₅₀ ≤ 25 mg/tissue: GHS Category 3 or 4 and EC₂₅/₅₀ > 25 mg/tissue: No Category. The direct application method showed an excellent predictive accuracy of 76.92 % (10/13). When 20 additional reference chemicals (33 total) were tested, the EC₅₀-based classification achieved a predictive accuracy of 75.76 % (25/33). Notably, applying a fixed-dose viability threshold (≤50 % viability at 5 mg/tissue for GHS 1/2; >50 % at 5 and ≤ 50 % at 25 mg/tissue for GHS 3/4; others as No Category) yielded comparable predictive performance. Collectively, these results suggest that SoluAirway™ ARTT is a promising, cost-effective alternative method for assessing acute inhalation toxicity.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106175"},"PeriodicalIF":2.7,"publicationDate":"2025-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145558319","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 : 2025-11-11DOI: 10.1016/j.tiv.2025.106174
Bulbul Ahmed , Melissa G. Farb , Kimberly Chen , Luise Pernar , Brain Carmine , Donald T. Hess , Jennifer Schlezinger , Noyan Gokce
p,p’-Dichlorodiphenyldichloroethylene (p,p’-DDE) is a primary and persistent metabolite of the pesticide dichlorodiphenyltrichloroethane. p,p’-DDE bioaccumulates in humans and has been associated with adverse health effects including endocrine disruption and cancer. Epidemiological studies also suggest associations with cardiovascular disease, but mechanisms are unknown. Here, we sought to investigate the effects of p,p’-DDE on human angiogenic microvascular function. Blood plasma samples (2006–2021, n = 80) and subcutaneous adipose tissues specimens (2024–2025, n = 18) were collected from adult human subjects undergoing elective bariatric surgery. Plasma samples were analyzed for p,p’-dichlorodiphenyltrichloroethane (p,p’-DDT) and its primary metabolites p,p’-dichlorodiphenyldichloroethane (p,p’-DDD) and p,p’-DDE by GC–MS. In a separate cohort of bariatric subjects, we examined the effect of p,p’-DDE on angiogenic capacity in fat pad specimens ex vivo. Cultured human aortic endothelial cells (HAECs) were used to study p,p’-DDE-mediated transcriptional changes in endothelial cells. p,p’-DDE was detected in the blood of all subjects and at higher concentrations than p,p’-DDD or p,p’-DDT. Ex vivo p,p’-DDE exposure significantly impaired microvascular angiogenic capacity (P < 0.05 vs. control) in adipose tissue and downregulated fibroblast growth factor 2 (FGF2) gene expression in HAECs in a dose-dependent manner. Moreover, we observed that p,p’-DDE-induced angiogenic impairment was fully reversed by exogenous recombinant FGF2 supplementation. We provide evidence that p,p’-DDE may interfere with human microvascular angiogenic processes via downregulation of FGF2 expression. Our results provide a novel mechanism by which environmental pollutants such as p,p’-DDE could negatively impact cardiovascular health.
{"title":"Anti-angiogenic effects of p,p’-Dichlorodiphenyldichloroethylene (p,p’-DDE) in the human microvasculature","authors":"Bulbul Ahmed , Melissa G. Farb , Kimberly Chen , Luise Pernar , Brain Carmine , Donald T. Hess , Jennifer Schlezinger , Noyan Gokce","doi":"10.1016/j.tiv.2025.106174","DOIUrl":"10.1016/j.tiv.2025.106174","url":null,"abstract":"<div><div>p,p’-Dichlorodiphenyldichloroethylene (p,p’<strong>-</strong>DDE) is a primary and persistent metabolite of the pesticide dichlorodiphenyltrichloroethane. p,p’<strong>-</strong>DDE bioaccumulates in humans and has been associated with adverse health effects including endocrine disruption and cancer. Epidemiological studies also suggest associations with cardiovascular disease, but mechanisms are unknown. Here, we sought to investigate the effects of p,p’<strong>-</strong>DDE on human angiogenic microvascular function. Blood plasma samples (2006–2021, <em>n</em> = 80) and subcutaneous adipose tissues specimens (2024–2025, <em>n</em> = 18) were collected from adult human subjects undergoing elective bariatric surgery. Plasma samples were analyzed for p,p’-dichlorodiphenyltrichloroethane (p,p’-DDT) and its primary metabolites p,p’-dichlorodiphenyldichloroethane (p,p’-DDD) and p,p’<strong>-</strong>DDE by GC–MS. In a separate cohort of bariatric subjects, we examined the effect of p,p’<strong>-</strong>DDE on angiogenic capacity in fat pad specimens <em>ex vivo</em>. Cultured human aortic endothelial cells (HAECs) were used to study p,p’<strong>-</strong>DDE-mediated transcriptional changes in endothelial cells. p,p’<strong>-</strong>DDE was detected in the blood of all subjects and at higher concentrations than p,p’-DDD or p,p’-DDT. <em>Ex vivo</em> p,p’<strong>-</strong>DDE exposure significantly impaired microvascular angiogenic capacity (<em>P</em> < 0.05 <em>vs.</em> control) in adipose tissue and downregulated fibroblast growth factor 2 (FGF2) gene expression in HAECs in a dose-dependent manner. Moreover, we observed that p,p’<strong>-</strong>DDE-induced angiogenic impairment was fully reversed by exogenous recombinant FGF2 supplementation. We provide evidence that p,p’<strong>-</strong>DDE may interfere with human microvascular angiogenic processes <em>via</em> downregulation of FGF2 expression. Our results provide a novel mechanism by which environmental pollutants such as p,p’<strong>-</strong>DDE could negatively impact cardiovascular health.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106174"},"PeriodicalIF":2.7,"publicationDate":"2025-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145514946","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}
Occupational exposure to metal-containing particles can pose significant risks to skin health, particularly in environments where materials such as stainless steel and cement are handled. This study evaluates the effectiveness of simple aqueous cleansing methods—water and soapy water—for removing metals from the skin after exposure to stainless steel particles (SSP) and cement dust, with the aim of informing dermal decontamination practices. Using an in vitro skin permeation model with Franz diffusion cells, we assessed the extent of metal absorption following exposure, with decontamination applied 15 min post-contact. Quantification of permeated metals was performed via ICP-MS. Notably, soapy water significantly reduced dermal retention of lead (Pb) from cement dust, from 305 ± 111 ng/cm2 (untreated) to 7.74 ± 0.7 ng/cm2 (p < 0.05). Conversely, the same procedure increased skin retention of nickel (Ni) and chromium (Cr), suggesting a wash-in effect. For SSP exposure, soapy water was effective in reducing permeation of all tested metals. These findings emphasize the need for tailored skin cleansing strategies and contribute to the development of protective and cleansing formulations aimed at mitigating metal-induced skin toxicity.
{"title":"Evaluation of metal removal from skin exposed to stainless steel and cement particles: Insights from an in vitro permeation study","authors":"Greta Camilla Magnano , Veronique Malard , Matteo Crosera , Giovanna Marussi , Olivier Debellemanière , Marcella Mauro , Francesca Larese Filon","doi":"10.1016/j.tiv.2025.106173","DOIUrl":"10.1016/j.tiv.2025.106173","url":null,"abstract":"<div><div>Occupational exposure to metal-containing particles can pose significant risks to skin health, particularly in environments where materials such as stainless steel and cement are handled. This study evaluates the effectiveness of simple aqueous cleansing methods—water and soapy water—for removing metals from the skin after exposure to stainless steel particles (SSP) and cement dust, with the aim of informing dermal decontamination practices. Using an in vitro skin permeation model with Franz diffusion cells, we assessed the extent of metal absorption following exposure, with decontamination applied 15 min post-contact. Quantification of permeated metals was performed via ICP-MS. Notably, soapy water significantly reduced dermal retention of lead (Pb) from cement dust, from 305 ± 111 ng/cm<sup>2</sup> (untreated) to 7.74 ± 0.7 ng/cm<sup>2</sup> (<em>p</em> < 0.05). Conversely, the same procedure increased skin retention of nickel (Ni) and chromium (Cr), suggesting a wash-in effect. For SSP exposure, soapy water was effective in reducing permeation of all tested metals. These findings emphasize the need for tailored skin cleansing strategies and contribute to the development of protective and cleansing formulations aimed at mitigating metal-induced skin toxicity.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106173"},"PeriodicalIF":2.7,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145483968","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}
In recent years, increasing attention has been directed toward development of in vitro methods to assess thyroid hormone system disrupting chemicals suitable for regulatory applications. As a part of this effort, the EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM) coordinated an international validation project through the European Union Network of Laboratories for the Validation of Alternative Methods (EU-NETVAL). This initiative involved the validation of 18 test methods, all aimed at contributing to a comprehensive testing battery for thyroid disruption. This work presents our contribution to the project, specifically focusing on the further development of a method designed to screen for disruption of the human thyrotropin receptor (hTSHR). Our objective is to provide an update on recent advancements in creating a reliable in vitro assay for toxicological assessment of chemicals that mimic or inhibit thyroid-stimulating hormone (TSH) function. We present and discuss pilot data generated using the developed standard operating procedure, including results from the initial testing of coded chemicals. The work highlights key challenges encountered during the method development and offers recommendations for future research and improvement. Our aim is to encourage more research teams to build upon this foundation, contributing to the refinement of a robust and reliable method. Ultimately, this approach could support the development of a comprehensive test battery for detecting disruptions in thyroid hormone signaling.
{"title":"Advances in thyroid hormone system disruption testing – In vitro method for monitoring disrupted thyrotropin signaling through the human TSH receptor (hTSHR)","authors":"Markéta Dvořáková, Kristina Kejlová, Gabriela Goffová, Eliška Pácalová, Lada Svobodová, Dagmar Jírová","doi":"10.1016/j.tiv.2025.106171","DOIUrl":"10.1016/j.tiv.2025.106171","url":null,"abstract":"<div><div>In recent years, increasing attention has been directed toward development of <em>in vitro</em> methods to assess thyroid hormone system disrupting chemicals suitable for regulatory applications. As a part of this effort, the EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM) coordinated an international validation project through the European Union Network of Laboratories for the Validation of Alternative Methods (EU-NETVAL). This initiative involved the validation of 18 test methods, all aimed at contributing to a comprehensive testing battery for thyroid disruption. This work presents our contribution to the project, specifically focusing on the further development of a method designed to screen for disruption of the human thyrotropin receptor (hTSHR). Our objective is to provide an update on recent advancements in creating a reliable <em>in vitro</em> assay for toxicological assessment of chemicals that mimic or inhibit thyroid-stimulating hormone (TSH) function. We present and discuss pilot data generated using the developed standard operating procedure, including results from the initial testing of coded chemicals. The work highlights key challenges encountered during the method development and offers recommendations for future research and improvement. Our aim is to encourage more research teams to build upon this foundation, contributing to the refinement of a robust and reliable method. Ultimately, this approach could support the development of a comprehensive test battery for detecting disruptions in thyroid hormone signaling.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106171"},"PeriodicalIF":2.7,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145432968","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 : 2025-10-30DOI: 10.1016/j.tiv.2025.106170
Pinzheng Li , Hengbin Zhang , Yingchun Chen , Shuanghui Lu , Huidi Jiang , Su Zeng , Hui Zhou
Oxoglaucine (OXO), an aporphine alkaloid derived from Corydalis yanhusuo, exhibits a broad spectrum of pharmacological activities, including antiviral, antifungal, antiplatelet, and anti-hepatic fibrosis effects, with particularly promising anticancer potential. In this study, we systematically investigated the hepatotoxicity profile and underlying mechanisms of OXO using in vitro models. Dose–response assays revealed that while OXO induced significant cytotoxicity in hepatocytes, primary hepatocytes exhibited markedly reduced sensitivity compared to hepatocellular carcinoma cells at therapeutically relevant concentrations. Mechanistic studies in mouse primary hepatocytes and human primary hepatocytes attributed this selective cytotoxicity to differential cytochrome P450 (CYP) enzyme expression. This finding was functionally validated by the observation that co-treatment with the broad-spectrum CYP inhibitor aminobenzotriazole (ABT) enhanced OXO-induced toxicity in mouse primary hepatocytes and human primary hepatocytes. Regarding its mechanism of toxicity, OXO induced marked cytotoxicity in the HepG2 cell line by triggering mitochondrial-mediated apoptosis, as evidenced by a decreased BCL2/BAX ratio, cytochrome c (CYCS) release, caspase-3 (CASP3) activation, and S-phase cell cycle arrest. Collectively, our findings in HepG2 cells and primary hepatocytes elucidate the role of CYP-mediated metabolism in the selective cytotoxicity of OXO. These findings not only provide crucial mechanistic insights but also support the further development of OXO as a promising candidate for preclinical and clinical evaluation in hepatocellular carcinoma.
{"title":"CYP-mediated metabolic divergence underpins oxoglaucine selectivity: Detoxification in healthy hepatocytes versus mitochondrial apoptosis in hepatocellular carcinoma","authors":"Pinzheng Li , Hengbin Zhang , Yingchun Chen , Shuanghui Lu , Huidi Jiang , Su Zeng , Hui Zhou","doi":"10.1016/j.tiv.2025.106170","DOIUrl":"10.1016/j.tiv.2025.106170","url":null,"abstract":"<div><div>Oxoglaucine (OXO), an aporphine alkaloid derived from <em>Corydalis yanhusuo</em>, exhibits a broad spectrum of pharmacological activities, including antiviral, antifungal, antiplatelet, and anti-hepatic fibrosis effects, with particularly promising anticancer potential. In this study, we systematically investigated the hepatotoxicity profile and underlying mechanisms of OXO using in vitro models. Dose–response assays revealed that while OXO induced significant cytotoxicity in hepatocytes, primary hepatocytes exhibited markedly reduced sensitivity compared to hepatocellular carcinoma cells at therapeutically relevant concentrations. Mechanistic studies in mouse primary hepatocytes and human primary hepatocytes attributed this selective cytotoxicity to differential cytochrome P450 (CYP) enzyme expression. This finding was functionally validated by the observation that co-treatment with the broad-spectrum CYP inhibitor aminobenzotriazole (ABT) enhanced OXO-induced toxicity in mouse primary hepatocytes and human primary hepatocytes. Regarding its mechanism of toxicity, OXO induced marked cytotoxicity in the HepG2 cell line by triggering mitochondrial-mediated apoptosis, as evidenced by a decreased BCL2/BAX ratio, cytochrome c (CYCS) release, caspase-3 (CASP3) activation, and S-phase cell cycle arrest. Collectively, our findings in HepG2 cells and primary hepatocytes elucidate the role of CYP-mediated metabolism in the selective cytotoxicity of OXO. These findings not only provide crucial mechanistic insights but also support the further development of OXO as a promising candidate for preclinical and clinical evaluation in hepatocellular carcinoma.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"111 ","pages":"Article 106170"},"PeriodicalIF":2.7,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145423432","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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