Pub Date : 2025-03-15DOI: 10.1016/j.tiv.2025.106053
Magali Araujo, Erica Stewart, Yang Zhao, Estatira Sepher, Cory Vaugh, Clara Erice, Robert L Sprando
Cannabinoids are highly lipophilic constituents of the hemp plant, which is present in several products intended for consumption. While cannabidiol (CBD) effects to humans have been extensively investigated, there is limited information on other minor cannabinoids. CBD oral bioavailability is low but increases with food and high-fat intake. We used Caco-2 cells in vitro to assess intestinal absorption of five cannabinoids (CBD, CBC, CBG, CBN, and CBDV) present in a CBD-rich hemp extract. We used a fed-state simulated intestinal fluid (Fessif) for dissolution of cannabinoids. Cannabinoids did not alter Caco-2 monolayer integrity. Except for CBC, recovery of cannabinoids decreased significantly after 90-minute incubation, compared to 60-minute incubation. No measurable cannabinoids were identified in the bottom chambers. Recovery of CBD, CBC, CBG and CBN after incubation with hemp extract or cannabinoid mix containing 30 μM CBD was unchanged, but CBDV recovery decreased. With hemp extract or a mix containing 10 μM CBD, recovery of CBD and CBC did not change, CBG recovery was lower (80-82 %), and CBN and CBDV were unquantifiable. This study highlights the challenges of evaluating permeability of cannabinoids by Caco-2 cells to predict intestinal absorption, including the physicochemical properties of these compounds, incubation time and cell properties.
{"title":"Assessment of intestinal absorption of five cannabinoids from an ethanolic CBD-rich hemp extract using Caco-2 cells in vitro.","authors":"Magali Araujo, Erica Stewart, Yang Zhao, Estatira Sepher, Cory Vaugh, Clara Erice, Robert L Sprando","doi":"10.1016/j.tiv.2025.106053","DOIUrl":"https://doi.org/10.1016/j.tiv.2025.106053","url":null,"abstract":"<p><p>Cannabinoids are highly lipophilic constituents of the hemp plant, which is present in several products intended for consumption. While cannabidiol (CBD) effects to humans have been extensively investigated, there is limited information on other minor cannabinoids. CBD oral bioavailability is low but increases with food and high-fat intake. We used Caco-2 cells in vitro to assess intestinal absorption of five cannabinoids (CBD, CBC, CBG, CBN, and CBDV) present in a CBD-rich hemp extract. We used a fed-state simulated intestinal fluid (Fessif) for dissolution of cannabinoids. Cannabinoids did not alter Caco-2 monolayer integrity. Except for CBC, recovery of cannabinoids decreased significantly after 90-minute incubation, compared to 60-minute incubation. No measurable cannabinoids were identified in the bottom chambers. Recovery of CBD, CBC, CBG and CBN after incubation with hemp extract or cannabinoid mix containing 30 μM CBD was unchanged, but CBDV recovery decreased. With hemp extract or a mix containing 10 μM CBD, recovery of CBD and CBC did not change, CBG recovery was lower (80-82 %), and CBN and CBDV were unquantifiable. This study highlights the challenges of evaluating permeability of cannabinoids by Caco-2 cells to predict intestinal absorption, including the physicochemical properties of these compounds, incubation time and cell properties.</p>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":" ","pages":"106053"},"PeriodicalIF":2.6,"publicationDate":"2025-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143652047","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 : 2025-03-14DOI: 10.1016/j.tiv.2025.106055
Jeong-hyun Hong , Kyung-Min Lim
Skin corrosion and irritation are key local toxicological responses of the skin to chemical exposure. Conventional in vivo methods have limitations such as species difference, ethical concern, and reproducibility issue, necessitating the development of reliable alternatives. We developed skin corrosion (SCT) and irritation test (SIT) methods using ex vivo porcine ear skin based on cell viability and skin barrier disruption. Test chemicals were treated on ex vivo porcine ear skin for 3 min and 60 min (SCT) or 60 min only (SIT). After 40 ± 2 h post-incubation, cell viability assay with CCK-8, and skin barrier test with FITC-dextran penetration were conducted. To evaluate the predictive capacity of the method, 38 reference chemicals (20 corrosives, 8 irritants and 10 no category chemicals) were tested. SCT achieved 96.7 % (29/30) accuracy in identifying corrosives, meeting the performance standards of OECD TG 431 In Vitro Skin Corrosion: Reconstructed Human Epidermis test. The accuracy for subcategorizing corrosive substances into categories 1 A (<25 % at 3 min) and 1B/1C (≥25 % at 3 min, <25 % at 60 min) was 83.3 % (25/30). The accuracy of SIT identifying non-irritant (≥50 % at 60 min) was 83.3 % (15/18). FITC-dextran penetration assay showed a similar accuracy, highlighting its value as an alternative endpoint to identify irritants. Collectively, this study demonstrated that the ex vivo porcine skin model may offer a cost-effective, and reliable alternative for skin hazard testing.
{"title":"Development of skin corrosion and irritation test methods employing ex vivo porcine skin model","authors":"Jeong-hyun Hong , Kyung-Min Lim","doi":"10.1016/j.tiv.2025.106055","DOIUrl":"10.1016/j.tiv.2025.106055","url":null,"abstract":"<div><div>Skin corrosion and irritation are key local toxicological responses of the skin to chemical exposure. Conventional <em>in vivo</em> methods have limitations such as species difference, ethical concern, and reproducibility issue, necessitating the development of reliable alternatives. We developed skin corrosion (SCT) and irritation test (SIT) methods using <em>ex vivo</em> porcine ear skin based on cell viability and skin barrier disruption. Test chemicals were treated on <em>ex vivo</em> porcine ear skin for 3 min and 60 min (SCT) or 60 min only (SIT). After 40 ± 2 h post-incubation, cell viability assay with CCK-8, and skin barrier test with FITC-dextran penetration were conducted. To evaluate the predictive capacity of the method, 38 reference chemicals (20 corrosives, 8 irritants and 10 no category chemicals) were tested. SCT achieved 96.7 % (29/30) accuracy in identifying corrosives, meeting the performance standards of OECD TG 431 <em>In Vitro</em> Skin Corrosion: Reconstructed Human Epidermis test. The accuracy for subcategorizing corrosive substances into categories 1 A (<25 % at 3 min) and 1B/1C (≥25 % at 3 min, <25 % at 60 min) was 83.3 % (25/30). The accuracy of SIT identifying non-irritant (≥50 % at 60 min) was 83.3 % (15/18). FITC-dextran penetration assay showed a similar accuracy, highlighting its value as an alternative endpoint to identify irritants. Collectively, this study demonstrated that the <em>ex vivo</em> porcine skin model may offer a cost-effective, and reliable alternative for skin hazard testing.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106055"},"PeriodicalIF":2.6,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639850","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 : 2025-03-13DOI: 10.1016/j.tiv.2025.106054
Shanshan Wang , Dongqian Guo , Xian Chen , Su-Zhu Chen , Xi-Wen Cui , Yong-He Han , Ping Xiang
Antimony (Sb) ore exploitation and the use of Sb-containing drugs pose known health risks. This study investigated the toxicity of environmentally relevant concentrations of Sb (0.12–12 mg L−1) on human umbilical vein endothelial cells (HUVECs). The 50 % lethal concentration (LC50) of Sb to HUVECs was 11.4 mg L−1. Exposing to high level of Sb induced cell cycle arrest by altering the expression of cell cycle regulators, inhibiting the transitions of G0/G1 to S and S to G2/M. At 1.2 mg L−1 Sb, CKD6 and p21 expressions in HUVECs changed to 0.75 and 1.32 folds that of no-Sb control, respectively (p < 0.01). At 12 mg L−1 Sb, CDK2, CKD6, and p27 expressions decreased by 1.54, 4.41, and 1.54 folds (p < 0.001), while p21 expression increased by 3.03 folds (p < 0.001) as compared to control. Sb also led to cell apoptosis, evidenced by Annexin V-FITC/PI staining and changes in the expressions of Bax (1.21–1.30 folds, p < 0.01) and Bcl-2 (0.65–0.83 folds). Oxidative damage was a pivotal factor driving cell apoptosis, probably through down-regulating antioxidant genes (CAT, GPX1, and GSTP1) and up-regulating stress response genes (HO-1, SOD1, and TrxR1). The elevated H2O2 generated in mitochondria likely contributed to cell apoptosis due to the imbalance in H2O2 metabolism. These findings suggest that environmentally relevant concentrations of Sb can exert cytotoxicity to HUVECs, which should be of potential concern for human cardiovascular disease.
{"title":"Environmentally relevant concentrations of antimony pose potential risks to human health: An evaluation on human umbilical vein endothelial cells","authors":"Shanshan Wang , Dongqian Guo , Xian Chen , Su-Zhu Chen , Xi-Wen Cui , Yong-He Han , Ping Xiang","doi":"10.1016/j.tiv.2025.106054","DOIUrl":"10.1016/j.tiv.2025.106054","url":null,"abstract":"<div><div>Antimony (Sb) ore exploitation and the use of Sb-containing drugs pose known health risks. This study investigated the toxicity of environmentally relevant concentrations of Sb (0.12–12 mg L<sup>−1</sup>) on human umbilical vein endothelial cells (HUVECs). The 50 % lethal concentration (LC<sub>50</sub>) of Sb to HUVECs was 11.4 mg L<sup>−1</sup>. Exposing to high level of Sb induced cell cycle arrest by altering the expression of cell cycle regulators, inhibiting the transitions of G<sub>0</sub>/G<sub>1</sub> to S and S to G<sub>2</sub>/M. At 1.2 mg L<sup>−1</sup> Sb, <em>CKD6</em> and <em>p21</em> expressions in HUVECs changed to 0.75 and 1.32 folds that of no-Sb control, respectively (<em>p</em> < 0.01). At 12 mg L<sup>−1</sup> Sb, <em>CDK2</em>, <em>CKD6</em>, and <em>p27</em> expressions decreased by 1.54, 4.41, and 1.54 folds (<em>p</em> < 0.001), while <em>p21</em> expression increased by 3.03 folds (<em>p</em> < 0.001) as compared to control. Sb also led to cell apoptosis, evidenced by Annexin V-FITC/PI staining and changes in the expressions of <em>Bax</em> (1.21–1.30 folds, <em>p</em> < 0.01) and <em>Bcl-2</em> (0.65–0.83 folds). Oxidative damage was a pivotal factor driving cell apoptosis, probably through down-regulating antioxidant genes (<em>CAT</em>, <em>GPX1</em>, and <em>GSTP1</em>) and up-regulating stress response genes (<em>HO-1</em>, <em>SOD1</em>, and <em>TrxR1</em>). The elevated H<sub>2</sub>O<sub>2</sub> generated in mitochondria likely contributed to cell apoptosis due to the imbalance in H<sub>2</sub>O<sub>2</sub> metabolism. These findings suggest that environmentally relevant concentrations of Sb can exert cytotoxicity to HUVECs, which should be of potential concern for human cardiovascular disease.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106054"},"PeriodicalIF":2.6,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628996","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 : 2025-03-13DOI: 10.1016/j.tiv.2025.106052
Sinenhlanhla X H Mthembu, Sithandiwe E Mazibuko-Mbeje, Sonia Silvestri, Patrick Orlando, Bongani B Nkambule, Christo J F Muller, Luca Tiano, Phiwayinkosi V Dludla
This study investigates the effects of prolonged simvastatin exposure on coenzyme Q9/10 (CoQ9/10) levels, an essential component of antioxidant defense, in cultured cardiac cells. Statins, commonly used to manage dyslipidemia and reduce cardiovascular risk, may impair mitochondrial function, but their impact on CoQ10 depletion and oxidative stress is not well understood. We examined the influence of simvastatin on mitochondrial oxidative capacity, reactive oxygen species (ROS) production, and CoQ9/10 status at concentrations of 0.3, 0.6, 1.25, 2.5, 5, 10, and 20 μM, over durations of 24, 48, and 72 h. Using an in vitro model of cultured H9c2 cardiomyoblasts, our results showed that short-term exposure (24 h) at lower concentrations (<5 μM) enhanced cytosolic and mitochondrial ROS levels without affecting mitochondrial function or CoQ9/10 status. However, prolonged exposure to higher concentrations (≥10 μM for >48 h) resulted in impaired mitochondrial oxidative capacity, indicated by increased proton leak and elevated ROS levels, which were followed by significantly reduced cell viability. These findings suggest that prolonged, high-dose simvastatin exposure may disrupt the oxidative balance of CoQ9/10, leading to myocardial injury. This research addresses a gap in understanding the long-term effects of statins on mitochondrial health and underscores the need for further studies to optimize statin therapy and minimize adverse effects on myocardial function.
{"title":"Prolonged exposure to simvastatin affects coenzyme Q<sub>9/10</sub> status leading to impaired mitochondrial respiratory capacity and reduced viability of cultured cardiac cells.","authors":"Sinenhlanhla X H Mthembu, Sithandiwe E Mazibuko-Mbeje, Sonia Silvestri, Patrick Orlando, Bongani B Nkambule, Christo J F Muller, Luca Tiano, Phiwayinkosi V Dludla","doi":"10.1016/j.tiv.2025.106052","DOIUrl":"https://doi.org/10.1016/j.tiv.2025.106052","url":null,"abstract":"<p><p>This study investigates the effects of prolonged simvastatin exposure on coenzyme Q<sub>9/10</sub> (CoQ<sub>9/10</sub>) levels, an essential component of antioxidant defense, in cultured cardiac cells. Statins, commonly used to manage dyslipidemia and reduce cardiovascular risk, may impair mitochondrial function, but their impact on CoQ<sub>10</sub> depletion and oxidative stress is not well understood. We examined the influence of simvastatin on mitochondrial oxidative capacity, reactive oxygen species (ROS) production, and CoQ<sub>9/10</sub> status at concentrations of 0.3, 0.6, 1.25, 2.5, 5, 10, and 20 μM, over durations of 24, 48, and 72 h. Using an in vitro model of cultured H9c2 cardiomyoblasts, our results showed that short-term exposure (24 h) at lower concentrations (<5 μM) enhanced cytosolic and mitochondrial ROS levels without affecting mitochondrial function or CoQ<sub>9/10</sub> status. However, prolonged exposure to higher concentrations (≥10 μM for >48 h) resulted in impaired mitochondrial oxidative capacity, indicated by increased proton leak and elevated ROS levels, which were followed by significantly reduced cell viability. These findings suggest that prolonged, high-dose simvastatin exposure may disrupt the oxidative balance of CoQ<sub>9/10</sub>, leading to myocardial injury. This research addresses a gap in understanding the long-term effects of statins on mitochondrial health and underscores the need for further studies to optimize statin therapy and minimize adverse effects on myocardial function.</p>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":" ","pages":"106052"},"PeriodicalIF":2.6,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143634996","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 : 2025-03-12DOI: 10.1016/j.tiv.2025.106050
Muhammed Yayla , Erdem Toktay , Bengul Ozdemir Sarikaya , Ugur Ermis , Sakir Akgun , Irfan Cinar
Purpose
The physiological effects of 5-HT7 receptors expressed in pancreatic beta cells have not yet been elucidated. We first aimed to investigate the effect of 5-HT7 receptor agonist (AS19) and antagonist (SB269970) application on insulin secretion in RIN-5F pancreatic beta cells. Subsequently, we aimed to investigate the effects of agonist and antagonist applications on cell damage induced by STZ.
Materials/Methods
Cell damage was caused by giving 5 mM STZ solution to the cells for 12 h. The protective effects of 5-HT7 receptor agonist and antagonist on this subsequent damage were investigated. IGF-1, TNF-α, TGF-B1, NF-KB, Bax, Caspase 3, Caspase 9, 5-HT7, 5HT7x2, 5HT7x3 mRNA expression levels were compared between groups.
Results
While agonist application stimulates insulin secretion, the effect of the antagonist varies.SB269970 reduced oxidative stress and downregulated TNF-a, TGF-B1 and NF-KB expression and also prevented apoptosis by decreasing Bax, caspase 3 and 9 levels against STZ-induced beta cell damage.
Conclusions
The effect of 5-HT7 receptors on insulin secretion and their effects against STZ damage will be guiding for more detailed studies in the treatment of diabetes and related diseases in the future.
{"title":"Effect of serotonin 7 receptors and transcript variants on RIN-5F pancreatic beta cell line","authors":"Muhammed Yayla , Erdem Toktay , Bengul Ozdemir Sarikaya , Ugur Ermis , Sakir Akgun , Irfan Cinar","doi":"10.1016/j.tiv.2025.106050","DOIUrl":"10.1016/j.tiv.2025.106050","url":null,"abstract":"<div><h3>Purpose</h3><div>The physiological effects of 5-HT7 receptors expressed in pancreatic beta cells have not yet been elucidated. We first aimed to investigate the effect of 5-HT7 receptor agonist (AS19) and antagonist (SB269970) application on insulin secretion in RIN-5F pancreatic beta cells. Subsequently, we aimed to investigate the effects of agonist and antagonist applications on cell damage induced by STZ.</div></div><div><h3>Materials/Methods</h3><div>Cell damage was caused by giving 5 mM STZ solution to the cells for 12 h. The protective effects of 5-HT7 receptor agonist and antagonist on this subsequent damage were investigated. IGF-1, TNF-α, TGF-B1, NF-KB, Bax, Caspase 3, Caspase 9, 5-HT7, 5HT7x2, 5HT7x3 mRNA expression levels were compared between groups.</div></div><div><h3>Results</h3><div>While agonist application stimulates insulin secretion, the effect of the antagonist varies.SB269970 reduced oxidative stress and downregulated TNF-a, TGF-B1 and NF-KB expression and also prevented apoptosis by decreasing Bax, caspase 3 and 9 levels against STZ-induced beta cell damage.</div></div><div><h3>Conclusions</h3><div>The effect of 5-HT7 receptors on insulin secretion and their effects against STZ damage will be guiding for more detailed studies in the treatment of diabetes and related diseases in the future.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106050"},"PeriodicalIF":2.6,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628997","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 : 2025-03-12DOI: 10.1016/j.tiv.2025.106040
Colleen M Pike, James A Levi, Lauren A Boone, Swetha Peddibhotla, Jacob Johnson, Bailey Zwarycz, Maureen K Bunger, William Thelin, Elizabeth M Boazak
Gastrointestinal toxicities (GITs) in clinical trials often lead to dose-limitations that reduce drug efficacy and delay treatment optimization. Preclinical animal models do not accurately replicate human physiology, leaving few options for early detection of GITs, such as diarrhea, before human studies. Chemotherapeutic agents, known to cause clinical diarrhea, frequently target mitotic cells. Therefore, we hypothesized a model utilizing proliferative cell populations derived from human intestinal crypts would predict clinical diarrhea occurrence with high accuracy. Here, we describe the development of a diarrhea prediction assay utilizing RepliGut® Planar, a primary intestinal stem cell-derived platform. To evaluate the ability of this model to predict clinical diarrhea risk, we assessed toxicity of 30 marketed drugs by measuring cell proliferation (EdU incorporation), cell abundance (nuclei quantification), and barrier function (TEER) in cells derived from three human donors. Dose response curves were generated for each drug, and the IC15 to Cmax ratio was used to identify a threshold for assay positivity. This model accurately predicted diarrhea potential, achieving an accuracy of 91 % for proliferation, 90 % for abundance, and 88 % for barrier formation. In vitro toxicity screening using primary proliferative cells may reduce clinical diarrhea and ultimately lead to safer and more effective treatments for patients.
{"title":"High-throughput assay for predicting diarrhea risk using a 2D human intestinal stem cell-derived model.","authors":"Colleen M Pike, James A Levi, Lauren A Boone, Swetha Peddibhotla, Jacob Johnson, Bailey Zwarycz, Maureen K Bunger, William Thelin, Elizabeth M Boazak","doi":"10.1016/j.tiv.2025.106040","DOIUrl":"10.1016/j.tiv.2025.106040","url":null,"abstract":"<p><p>Gastrointestinal toxicities (GITs) in clinical trials often lead to dose-limitations that reduce drug efficacy and delay treatment optimization. Preclinical animal models do not accurately replicate human physiology, leaving few options for early detection of GITs, such as diarrhea, before human studies. Chemotherapeutic agents, known to cause clinical diarrhea, frequently target mitotic cells. Therefore, we hypothesized a model utilizing proliferative cell populations derived from human intestinal crypts would predict clinical diarrhea occurrence with high accuracy. Here, we describe the development of a diarrhea prediction assay utilizing RepliGut® Planar, a primary intestinal stem cell-derived platform. To evaluate the ability of this model to predict clinical diarrhea risk, we assessed toxicity of 30 marketed drugs by measuring cell proliferation (EdU incorporation), cell abundance (nuclei quantification), and barrier function (TEER) in cells derived from three human donors. Dose response curves were generated for each drug, and the IC15 to Cmax ratio was used to identify a threshold for assay positivity. This model accurately predicted diarrhea potential, achieving an accuracy of 91 % for proliferation, 90 % for abundance, and 88 % for barrier formation. In vitro toxicity screening using primary proliferative cells may reduce clinical diarrhea and ultimately lead to safer and more effective treatments for patients.</p>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":" ","pages":"106040"},"PeriodicalIF":2.6,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143631048","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}
Zinc oxide nanoparticles (ZnONPs) and titanium dioxide nanoparticles (TiO2NPs), due to their nanometric size and photostability, are increasingly used as ingredients in sunscreens to absorb and scatter UV radiation. However, the current state of knowledge is insufficient to guarantee their safety. Therefore, the objectives of this study were to evaluate the cytotoxicity of ZnONPs and TiO2NPs in the presence and absence of UV radiation in in vitro model of primary human melanocyte cells HEMas. Our research demonstrated that 47 nm TiO2NPs exhibited lower toxicity compared to 25 nm ZnONPs. ZnONPs (5–12.5 ppm) affect various intracellular processes, including cell membrane integrity, proliferative processes, and the induction of morphological changes in cells at the ultrastructural level, particularly in mitochondria. The study highlights intricate mechanisms of cell death induced by ZnONPs, revealing a multifaceted interplay between apoptosis and necrosis. Additionally, we indicate the potential role of intracellular calcium ion influx, notably triggered by ZnONPs, in driving cell toxicity. This influx is linked to endoplasmic reticulum (ER) dysfunction, ultimately leading to cell death, offering valuable insights into the underlying mechanisms of nanoparticle-induced toxicity. Importantly, the co-exposure of both ZnONPs and TiO2NPs with UV radiation (9 J/cm2) enhances the toxic effect on melanocyte cells, indicating an interaction between NPs and UV radiaton and raising potential concerns about their effects on melanocytes and overall skin health.
{"title":"Cytotoxicity of ZnO nanoparticles in human melanocyte cells in the presence or absence of UV radiation: A preliminary comparative study with TiO2 nanoparticles","authors":"Karolina Niska , Beata Muszyńska , Szymon Kowalski , Emilia Tomaszewska , Magdalena Narajczyk , Monika Pawłowska , Paweł W. Majewski , Ewa Augustin , Iwona Inkielewicz-Stepniak","doi":"10.1016/j.tiv.2025.106051","DOIUrl":"10.1016/j.tiv.2025.106051","url":null,"abstract":"<div><div>Zinc oxide nanoparticles (ZnONPs) and titanium dioxide nanoparticles (TiO<sub>2</sub>NPs), due to their nanometric size and photostability, are increasingly used as ingredients in sunscreens to absorb and scatter UV radiation. However, the current state of knowledge is insufficient to guarantee their safety. Therefore, the objectives of this study were to evaluate the cytotoxicity of ZnONPs and TiO<sub>2</sub>NPs in the presence and absence of UV radiation in in vitro model of primary human melanocyte cells HEMas. Our research demonstrated that 47 nm TiO<sub>2</sub>NPs exhibited lower toxicity compared to 25 nm ZnONPs. ZnONPs (5–12.5 ppm) affect various intracellular processes, including cell membrane integrity, proliferative processes, and the induction of morphological changes in cells at the ultrastructural level, particularly in mitochondria. The study highlights intricate mechanisms of cell death induced by ZnONPs, revealing a multifaceted interplay between apoptosis and necrosis. Additionally, we indicate the potential role of intracellular calcium ion influx, notably triggered by ZnONPs, in driving cell toxicity. This influx is linked to endoplasmic reticulum (ER) dysfunction, ultimately leading to cell death, offering valuable insights into the underlying mechanisms of nanoparticle-induced toxicity. Importantly, the co-exposure of both ZnONPs and TiO<sub>2</sub>NPs with UV radiation (9 J/cm<sup>2</sup>) enhances the toxic effect on melanocyte cells, indicating an interaction between NPs and UV radiaton and raising potential concerns about their effects on melanocytes and overall skin health.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106051"},"PeriodicalIF":2.6,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628998","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}
Decomposing bodies release necro-leachate, a toxic fluid containing harmful compounds such as biogenic amines. This study investigated the genotoxic effects of the different concentrations (0.5, 1.4, 2.3, 3.2 mM) of bioamine putrescine on HepG2 cells using the comet assay, the micronucleus test, and gene expression analysis. The results were compared to negative control and indicated significant DNA damage in the comet assay highlighting tail DNA intensity that exhibited significant differences across all tested concentrations (0.5 = 192 %, 1.4 = 189 %, 2.3 = 208 %, 3.2 = 132 %). The micronucleus test revealed a significant increase in micronuclei for concentrations 0.5 (193 %), 1.4 (229 %), 2.3 (206 %); nuclear buds 3.2 (173 %); chromosomal bridges 3.2 (735 %). Furthermore, genes linked to oxidative stress and DNA damage exhibited statistically significant expression alterations. These findings suggest that putrescine has genotoxic potential in human-derived HepG2 cells, raising concerns about cemetery contaminants' occupational and environmental risks. This study is the first to assess putrescine's toxicity as an environmental pollutant, as previous research has mainly focused on its role in the food sector. These insights highlight the potential threats necro-leachate poses to environmental health, emphasizing the need for further research on cemetery pollution.
{"title":"Genotoxicity of putrescine and its effects on gene expression in HepG2 cell line","authors":"Franco Dani Campos-Pereira , Letícia Rocha Gonçalves, Raquel Vaz Hara Jardim, Letícia Bulascochi Cagnoni, Karen C.M. Moraes, Maria Aparecida Marin-Morales","doi":"10.1016/j.tiv.2025.106048","DOIUrl":"10.1016/j.tiv.2025.106048","url":null,"abstract":"<div><div>Decomposing bodies release necro-leachate, a toxic fluid containing harmful compounds such as biogenic amines. This study investigated the genotoxic effects of the different concentrations (0.5, 1.4, 2.3, 3.2 mM) of bioamine putrescine on HepG2 cells using the comet assay, the micronucleus test, and gene expression analysis. The results were compared to negative control and indicated significant DNA damage in the comet assay highlighting tail DNA intensity that exhibited significant differences across all tested concentrations (0.5 = 192 %, 1.4 = 189 %, 2.3 = 208 %, 3.2 = 132 %). The micronucleus test revealed a significant increase in micronuclei for concentrations 0.5 (193 %), 1.4 (229 %), 2.3 (206 %); nuclear buds 3.2 (173 %); chromosomal bridges 3.2 (735 %). Furthermore, genes linked to oxidative stress and DNA damage exhibited statistically significant expression alterations. These findings suggest that putrescine has genotoxic potential in human-derived HepG2 cells, raising concerns about cemetery contaminants' occupational and environmental risks. This study is the first to assess putrescine's toxicity as an environmental pollutant, as previous research has mainly focused on its role in the food sector. These insights highlight the potential threats necro-leachate poses to environmental health, emphasizing the need for further research on cemetery pollution.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106048"},"PeriodicalIF":2.6,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143631046","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 : 2025-03-10DOI: 10.1016/j.tiv.2025.106049
Julien Saint-Pol, Maxime Culot
Studies based on extracellular vesicles (EVs) have been multiplying exponentially for almost two decades, since they were first identified as vectors of cell-cell communication. However, several of these studies display a lack of rigor in EVs characterization and isolation, without discriminating between the different EV populations, thus generating conflicting and unreproducible results. There is therefore a strong need for standardization and guidelines to conduct studies that are rigorous, transparent, reproducible and comply with certain nomenclatures concerning the type of EVs used. The International Society for Extracellular Vesicles (ISEV) published the Minimum Information for Studies of Extracellular Vesicles (MISEV) in 2014, updating it in 2018 and 2023 to reflect different study contexts and technical advancements. The primary objective of this review is to inform future authors about EVs, including their history, nomenclature, and technical recommendations for the for isolation and functionality analysis for conducing EV-based studies according to current standards. Additionally, it aims to inform reviewers about the key parameters required for characterizing EV preparations.
{"title":"Minimum information for studies of extracellular vesicles (MISEV) as toolbox for rigorous, reproducible and homogeneous studies on extracellular vesicles","authors":"Julien Saint-Pol, Maxime Culot","doi":"10.1016/j.tiv.2025.106049","DOIUrl":"10.1016/j.tiv.2025.106049","url":null,"abstract":"<div><div>Studies based on extracellular vesicles (EVs) have been multiplying exponentially for almost two decades, since they were first identified as vectors of cell-cell communication. However, several of these studies display a lack of rigor in EVs characterization and isolation, without discriminating between the different EV populations, thus generating conflicting and unreproducible results. There is therefore a strong need for standardization and guidelines to conduct studies that are rigorous, transparent, reproducible and comply with certain nomenclatures concerning the type of EVs used. The International Society for Extracellular Vesicles (ISEV) published the Minimum Information for Studies of Extracellular Vesicles (MISEV) in 2014, updating it in 2018 and 2023 to reflect different study contexts and technical advancements. The primary objective of this review is to inform future authors about EVs, including their history, nomenclature, and technical recommendations for the for isolation and functionality analysis for conducing EV-based studies according to current standards. Additionally, it aims to inform reviewers about the key parameters required for characterizing EV preparations.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106049"},"PeriodicalIF":2.6,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143611146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-07DOI: 10.1016/j.tiv.2025.106047
Lexing Wei , Yang Chen , Minjia Wu , Peixuan Ma , Huan Wang , Yueming Jiang , Michael Aschner , Jing Zhou , Guodong Lu , Lina Zhao , Xiaowei Huang
Deltamethrin (DM), a widely used pyrethroid insecticide, has been increasingly recognized as a risk factor for neurodegeneration. However, the underlying mechanism is still far from clear. In this study, we investigated whether MEK1 is involved in DM-induced neurotoxicity and mediated mitochondrial-dependent apoptosis. In mouse hippocampal neuron HT22 cells model, DM (2,10,50 μM) dose-dependently increased apoptotic cells rate and impaired mitochondrial membrane potential (MMP), as well as significantly upregulated of apoptotic related proteins Bax, cytochrome c (Cyt-c) and Caspase-3 were observed. RNA-sequencing analysis further revealed that the MEK/ERK signal pathway was remarkably enriched and activated after DM exposure. In particularly, upregulation of MEK1, other than ERK1/2, was detected at both transcriptional and translational levels. Inhibition of MEK1 can effectively result in the recovery of mitochondrial morphology and MMP in DM-treated HT22 cells. And that further alleviated apoptosis by reversing the overexpression of Bax, Cyt-c and Caspase-3. Collectively, these findings demonstrate the critical role of MEK1 in regulating mitochondrial-dependent apoptosis induced by DM, providing a novel understanding of the neurotoxicity of DM.
{"title":"MEK1 inhibition ameliorates mitochondrial-dependent apoptosis induced by deltamethrin in mouse hippocampal neuron HT22 cells","authors":"Lexing Wei , Yang Chen , Minjia Wu , Peixuan Ma , Huan Wang , Yueming Jiang , Michael Aschner , Jing Zhou , Guodong Lu , Lina Zhao , Xiaowei Huang","doi":"10.1016/j.tiv.2025.106047","DOIUrl":"10.1016/j.tiv.2025.106047","url":null,"abstract":"<div><div>Deltamethrin (DM), a widely used pyrethroid insecticide, has been increasingly recognized as a risk factor for neurodegeneration. However, the underlying mechanism is still far from clear. In this study, we investigated whether MEK1 is involved in DM-induced neurotoxicity and mediated mitochondrial-dependent apoptosis. In mouse hippocampal neuron HT22 cells model, DM (2,10,50 μM) dose-dependently increased apoptotic cells rate and impaired mitochondrial membrane potential (MMP), as well as significantly upregulated of apoptotic related proteins Bax, cytochrome <em>c</em> (Cyt-c) and Caspase-3 were observed. RNA-sequencing analysis further revealed that the MEK/ERK signal pathway was remarkably enriched and activated after DM exposure. In particularly, upregulation of MEK1, other than ERK1/2, was detected at both transcriptional and translational levels. Inhibition of MEK1 can effectively result in the recovery of mitochondrial morphology and MMP in DM-treated HT22 cells. And that further alleviated apoptosis by reversing the overexpression of Bax, Cyt-c and Caspase-3. Collectively, these findings demonstrate the critical role of MEK1 in regulating mitochondrial-dependent apoptosis induced by DM, providing a novel understanding of the neurotoxicity of DM.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"106 ","pages":"Article 106047"},"PeriodicalIF":2.6,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143588223","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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