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A multi-omics approach reveals differences in toxicity and mechanisms in rice (Oryza sativa L.) exposed to anatase or rutile TiO2 nanoparticles. 多组学方法揭示了暴露于锐钛型或金红石型TiO2纳米粒子的水稻(Oryza sativa L.)在毒性和机理方面的差异。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-10-05 DOI: 10.1016/j.impact.2024.100530
Nan Zhao, Chang Liu, Chenyang Ji, Xuefeng Jiang, Jinyu Zhao, Liwen Qiang, Hangbiao Jin
{"title":"A multi-omics approach reveals differences in toxicity and mechanisms in rice (Oryza sativa L.) exposed to anatase or rutile TiO<sub>2</sub> nanoparticles.","authors":"Nan Zhao, Chang Liu, Chenyang Ji, Xuefeng Jiang, Jinyu Zhao, Liwen Qiang, Hangbiao Jin","doi":"10.1016/j.impact.2024.100530","DOIUrl":"10.1016/j.impact.2024.100530","url":null,"abstract":"<p><p>Titanium dioxide nanoparticles (TiO<sub>2</sub> NPs) have been widely used in agriculture, which increased the risk to soil-plant systems. Studies have demonstrated that TiO<sub>2</sub> NPs can induce phytotoxicity. However, the toxicity mechanisms, particularly under the stress of TiO<sub>2</sub> NPs with different crystalline forms, remain inadequately reported. In this study, we combined transcriptomics and metabolomics to analyze the toxicity mechanisms in rice (Oryza sativa L.) under the stress of anatase (AT) or rutile (RT) TiO<sub>2</sub> NPs (50 mg/kg, 40 days). The length (decreased by 1.1-fold, p = 0.021) and malondialdehyde concentration (decreased by 1.4-fold, p = 0.0027) of rice shoots was significantly reduced after AT exposure, while no significant changes were observed following RT exposure. Antioxidant enzyme activities were significantly altered both in the AT and RT groups, indicating TiO<sub>2</sub> NPs induced rice oxidative damage (with changes of 1.1 to 1.4-fold, p < 0.05). Additionally, compared to the control, AT exposure altered 3247 differentially expressed genes (DEGs) and 56 significantly differentially metabolites in rice (collectively involved in pyrimidine metabolism, TCA cycle, fatty acid metabolism, and amino acid metabolism). After RT exposure, 2814 DEGs and 55 significantly differentially metabolites were identified, which were collectively involved in fatty acid metabolism and amino acid metabolism. Our results indicated that AT exposure led to more pronounced changes in biological responses related to oxidative stress and had more negative effects on rice growth compared to RT exposure. These findings provide new insights into the phytotoxic mechanisms of TiO<sub>2</sub> NPs with different crystalline forms. Based on the observed adverse effects, the study emphasizes that any form of TiO<sub>2</sub> NPs should be used with caution in rice ecosystems. This study is the first to demonstrate that AT is more toxic than RT in paddy ecosystems, providing crucial insights into the differential impacts and toxic mechanisms of TiO<sub>2</sub> NPs with different crystalline forms. These findings suggest prioritizing the use of RT when TiO<sub>2</sub> NPs are necessary in agricultural development to minimize toxicity risks.</p>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142381274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Biodistribution and toxic potential of silver nanoparticles when introduced to the female rat reproductive tract. 银纳米粒子进入雌性大鼠生殖道时的生物分布和毒性潜力。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-09-21 DOI: 10.1016/j.impact.2024.100529
Nathan A Koonce, Ammu Mathew, Ioana-Mihaela Popescu, Kelly Davis, Doug Wagner, Gokulan Kuppan, Mugimane Manjanatha, Julian E A Leakey, Anil K Patri
{"title":"Biodistribution and toxic potential of silver nanoparticles when introduced to the female rat reproductive tract.","authors":"Nathan A Koonce, Ammu Mathew, Ioana-Mihaela Popescu, Kelly Davis, Doug Wagner, Gokulan Kuppan, Mugimane Manjanatha, Julian E A Leakey, Anil K Patri","doi":"10.1016/j.impact.2024.100529","DOIUrl":"10.1016/j.impact.2024.100529","url":null,"abstract":"<p><p>The prevalence of ionic silver and silver nanomaterials in hygiene products has been increasing due to their antimicrobial activity. While numerous studies have examined the effects of nanosilver in laboratory settings, there is a limited understanding of its impact on reproductive tissues, as well as its biodistribution and toxicity upon intra-vaginal exposure. If ionic or nanosilver enters adjacent and internal tissues via intra-vaginal exposure, the overuse of hygiene products containing silver may potentially threaten woman's health. This study investigated the effects of intra-vaginal silver exposure in Female Fischer 344 rats to single and multiple doses of a commercial product containing silver, along with standard nanosilver materials. Custom tampons were developed to simulate practical usage scenarios. The analysis of tissue biodistribution revealed that epithelial penetration and redistribution of silver was observed with most administered silver eliminated in feces (8-44 %), and secondary tissues containing 1-18 % of the dose, predominantly localized in the reproductive tract. In a subsequent toxicity study, vaginal histopathology indicated a cellular inflammatory reaction (neutrophil infiltration) associated with the presence of foreign silver material upon a single administration. Interestingly, no noticeable difference in histopathology incidence was observed upon multiple exposures to silver compared to the control group. Clinical chemistry and hematology analyses following acute exposure to silver nanomaterials showed no significant abnormalities. Overall, acute vaginal exposure to silver nanomaterials and ionic silver resulted in limited silver persistence, local tissue reactivity, epithelial penetration of silver resulting in accumulation in distant organs, and elimination primarily through feces. In vitro data suggested potential alterations in normal vaginal flora. Long-term studies are still lacking in this area.</p>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142308085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Toxicity and decomposition activity inhibition of VO2 micro/nanoparticles to white rot fungus Phanerochaete chrysosporium VO2 微/纳米粒子对白腐菌 Phanerochaete chrysosporium 的毒性和分解活性抑制作用。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-09-01 DOI: 10.1016/j.impact.2024.100528
{"title":"Toxicity and decomposition activity inhibition of VO2 micro/nanoparticles to white rot fungus Phanerochaete chrysosporium","authors":"","doi":"10.1016/j.impact.2024.100528","DOIUrl":"10.1016/j.impact.2024.100528","url":null,"abstract":"<div><p>Vanadium dioxide (VO<sub>2</sub>) is an excellent phase transition material widely used in various applications, and thus inevitably enters the environment via different routes and encounters various organisms. Nonetheless, limited information is available on the environmental hazards of VO<sub>2</sub>. In this study, we investigated the impact of two commercial VO<sub>2</sub> particles, nanosized S-VO<sub>2</sub> and micro-sized M-VO<sub>2</sub> on the white rot fungus <em>Phanerochaete chrysosporium</em>. The growth of <em>P. chrysosporium</em> is significantly affected by VO<sub>2</sub> particles, with S-VO<sub>2</sub> displaying a higher inhibitory effect on weight gain. In addition, VO<sub>2</sub> at high concentrations inhibits the formation of fungal fibrous hyphae and disrupts the integrity of fungus cells as evidenced by the cell membrane damage and the loss of cytoplasm. Notably, at 200 μg/mL, S-VO<sub>2</sub> completely alters the morphology of <em>P. chrysosporium</em>, while the M-VO<sub>2</sub> treatment does not affect the mycelium formation of <em>P. chrysosporium</em>. Additionally, VO<sub>2</sub> particles inhibit the laccase activity secreted by <em>P. chrysosporium</em>, and thus prevent the dye decoloration and sawdust decomposition by <em>P. chrysosporium</em>. The mechanism underlying this toxicity is related to the dissolution of VO<sub>2</sub> and the oxidative stress induced by VO<sub>2</sub>. Overall, our findings suggest that VO<sub>2</sub> nanoparticles pose significant environmental hazards and risks to white rot fungi.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142126161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Do nanoplastics impact Pb up-taking by Hordeum vulgare L.? 纳米塑料是否会影响 Hordeum vulgare L. 对铅的吸收?
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100526
{"title":"Do nanoplastics impact Pb up-taking by Hordeum vulgare L.?","authors":"","doi":"10.1016/j.impact.2024.100526","DOIUrl":"10.1016/j.impact.2024.100526","url":null,"abstract":"<div><p>Most studies on nanoplastics (NPs) focus on aquatic environments, overlooking their combined bioaccumulation with pollutants in terrestrial ecosystems. This study addresses a part of this gap by investigating how polystyrene nanoplastics (PS-NPs) affect the bioaccumulation and translocation of lead (Pb) in <em>Hordeum vulgare</em> L. plants. Using the RHIZOtest device for precise soil contamination control, we quantified PS-NPs (50 nm) in plant shoots via pyrolysis-gas chromatography/mass spectrometry (Py-GCMS) after plant KOH digestion. Our findings revealed that PS-NPs reduce Pb bioaccumulation and make adsorbed Pb onto PS-NPs less bioavailable to plants. For the highest Pb concentration, the Pb uptake index (PUI) followed the trend: Free Pb &gt; NPs + Pb &gt; Pb primary adsorbed by NPs, showing reduced Pb translocation to shoots in the presence of PS-NPs. Moreover, the presence of Pb decreased the bioavailability of PS-NPs probably in response to PS-NPs aggregation or modified charge. The PS-NPs concentrations in shoots range from 275.2 to 400 μg g<sup>−1</sup>, representing 3.9 to 5.75% of the total PS-NPs. This study highlights the intricate interactions between nanoplastics and metals in soil-plant systems and emphasizes the need for further research on their combined effects and potential risks to food safety.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141907069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Dancing with danger-how honeybees are getting affected in the web of microplastics-a review 与危险共舞--微塑料网如何影响蜜蜂--综述。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100522
{"title":"Dancing with danger-how honeybees are getting affected in the web of microplastics-a review","authors":"","doi":"10.1016/j.impact.2024.100522","DOIUrl":"10.1016/j.impact.2024.100522","url":null,"abstract":"<div><p>Anthropogenic activities have negatively impacted the ecosystem dramatically over the last few decades. The environment is becoming more contaminated with heavy metals, pesticides, and microplastics (MPs) as a result of the swift rise in industrialization and urbanisation. These contaminants are present everywhere in the ecosystem, affecting every living creature, from aquatic to terrestrial to aerial. Recently, the widespread of microplastics in the environment has raised serious concerns about the contamination of honey bees by these tiny particles of plastic. Honeybees are the major pollinators which contributes in the pollination of about 70% food that we consume. This review summarizes current research findings on the presence, uptake, and possible effects of microplastics on honey bees. Findings revealed the presence of microplastics in various honey bee matrices, such as honey, pollen, beeswax, and bee bodies, highlighting the potential routes of exposure for these vital pollinators. Additionally, evidence suggests that microplastics can accumulate in honey bee tissues (brain, midgut, Malpighian tubules, trachea, and haemolymph) potentially leading to adverse effects on honey bee health, behaviour, and colony dynamics. Additionally, MPs has a synergistic impact on immune system as well. Change in cuticle profile, reduction in body weight, and changes in eating frequency can regulate overall success rate of their survival. However, significant knowledge gaps remain regarding the long-term consequences for honey bee populations and ecosystem health, which cannot unveil the ultimate degree of future threats. Future research efforts should focus on investigating the interactions between microplastics and other stressors, such as pesticides and pathogens, and assessing the broader ecological implications of honey bee contamination with microplastics. Addressing these knowledge gaps is essential for developing effective mitigation strategies to minimize the impact of microplastics on honey bee populations and safeguarding their vital role in ecosystem functioning and food security.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141633957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Overview of in vitro-in vivo extrapolation approaches for the risk assessment of nanomaterial toxicity 纳米材料毒性风险评估的体外-体内外推法概述。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100524
{"title":"Overview of in vitro-in vivo extrapolation approaches for the risk assessment of nanomaterial toxicity","authors":"","doi":"10.1016/j.impact.2024.100524","DOIUrl":"10.1016/j.impact.2024.100524","url":null,"abstract":"<div><p>Nanomaterials are increasingly used in many applications due to their enhanced properties. To ensure their safety for humans and the environment, nanomaterials need to be evaluated for their potential risk. The risk assessment analysis on the nanomaterials based on animal or <em>in vivo</em> studies is accompanied by several concerns, including animal welfare, time and cost needed for the studies. Therefore, incorporating <em>in vitro</em> studies in the risk assessment process is increasingly considered. To be able to analyze the potential risk of nanomaterial to human health, there are factors to take into account. Utilizing <em>in vitro</em> data in the risk assessment analysis requires methods that can be used to translate <em>in vitro</em> data to predict <em>in vivo</em> phenomena (<em>in vitro</em>-<em>in vivo</em> extrapolation (IVIVE) methods) to be incorporated, to obtain a more accurate result. Apart from the experiments and species conversion (for example, translation between the cell culture, animal and human), the challenge also includes the unique properties of nanomaterials that might cause them to behave differently compared to the same materials in a bulk form. This overview presents the IVIVE techniques that are developed to extrapolate pharmacokinetics data or doses. A brief example of the IVIVE methods for chemicals is provided, followed by a more detailed summary of available IVIVE methods applied to nanomaterials. The IVIVE techniques discussed include the comparison between <em>in vitro</em> and <em>in vivo</em> studies, methods to rene the dose metric or the <em>in vitro</em> models, allometric approach, mechanistic modeling, Multiple-Path Particle Dosimetry (MPPD), methods using organ burden data and also approaches that are currently being developed.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141766729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A modified methodology for extraction and quantification of microplastics in soil 土壤中微塑料提取和定量的改进方法。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100525
{"title":"A modified methodology for extraction and quantification of microplastics in soil","authors":"","doi":"10.1016/j.impact.2024.100525","DOIUrl":"10.1016/j.impact.2024.100525","url":null,"abstract":"<div><p>The ubiquitousness of microplastics (&lt;5 mm) has become a pressing environmental concern globally due to the extensive use of plastics. Microplastics have been well-studied in aquatic environments but not well-characterized in soils. Present analytical processes to quantify microplastics accurately in soil samples are quite challenging and require improved and validated analytical steps to eliminate the obscurities and biases. We aimed to develop an effective method for the extraction and quantification of microplastics from soil samples. Different ratios of low-(NaCl) and high-density solutions (ZnCl<sub>2</sub>/ NaBr) were tested to determine the most efficient combination for density-dependent separation of microplastics from soil. The combination of low- (1:6) and high-density (1:3) solutions {as weight of soil(g)/volume of density solution(ml)} accounted for 95% recovery of the spiked microplastic particles from soil samples. Likewise, different soil-to-solution ratios of H<sub>2</sub>O<sub>2</sub> were tested for the removal of soil organic matter with heating and non-heating steps. Prior removal of organic matter from soil samples achieved a clear supernatant that facilitated 99% recovery of microplastic particles. The validation of individually spiked microplastic particles of small (10-100 μm) and large scale (100-5000 μm) resulted in recovery ranging from 88 to 99%. A validated modified method with prior digestion followed by density-dependent separation was further tested using the field samples with microplastic contamination. The microplastics of different shapes, sizes, colours and polymeric compositions were reported efficiently and well characterized in the field-collected soil samples using this method.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141971442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Next Generation Risk Assessment approaches for advanced nanomaterials: Current status and future perspectives 先进纳米材料的下一代风险评估方法:现状与未来展望。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100523
{"title":"Next Generation Risk Assessment approaches for advanced nanomaterials: Current status and future perspectives","authors":"","doi":"10.1016/j.impact.2024.100523","DOIUrl":"10.1016/j.impact.2024.100523","url":null,"abstract":"<div><p>This manuscript discusses the challenges of applying New Approach Methodologies (NAMs) for safe by design and regulatory risk assessment of advanced nanomaterials (AdNMs). The authors propose a framework for Next Generation Risk Assessment of AdNMs involving NAMs that is aligned to the conventional risk assessment paradigm. This framework is exposure-driven, endpoint-specific, makes best use of pre-existing information, and can be implemented in tiers of increasing specificity and complexity of the adopted NAMs. The tiered structure of the approach, which effectively combines the use of existing data with targeted testing will allow safety to be assessed cost-effectively and as far as possible with even more limited use of vertebrates. The regulatory readiness of state-of-the-art emerging NAMs is assessed in terms of Transparency, Reliability, Accessibility, Applicability, Relevance and Completeness, and their appropriateness for AdNMs is discussed in relation to each step of the risk assessment paradigm along with providing perspectives for future developments in the respective scientific and regulatory areas.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2452074824000338/pdfft?md5=b6feab203d19e28d8b0aafbc6ce5d6d7&pid=1-s2.0-S2452074824000338-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141766728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Improving the dichloro-dihydro-fluorescein (DCFH) assay for the assessment of intracellular reactive oxygen species formation by nanomaterials 改进二氯二氢荧光素(DCFH)测定法,以评估纳米材料在细胞内形成的活性氧。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100521
Nienke Ruijter , Margriet van der Zee , Alberto Katsumiti , Matthew Boyles , Flemming R. Cassee , Hedwig Braakhuis
{"title":"Improving the dichloro-dihydro-fluorescein (DCFH) assay for the assessment of intracellular reactive oxygen species formation by nanomaterials","authors":"Nienke Ruijter ,&nbsp;Margriet van der Zee ,&nbsp;Alberto Katsumiti ,&nbsp;Matthew Boyles ,&nbsp;Flemming R. Cassee ,&nbsp;Hedwig Braakhuis","doi":"10.1016/j.impact.2024.100521","DOIUrl":"10.1016/j.impact.2024.100521","url":null,"abstract":"<div><p>To facilitate Safe and Sustainable by Design (SSbD) strategies during the development of nanomaterials (NMs), quick and easy <em>in vitro</em> assays to test for hazard potential at an early stage of NM development are essential. The formation of reactive oxygen species (ROS) and the induction of oxidative stress are considered important mechanisms that can lead to NM toxicity. <em>In vitro</em> assays measuring oxidative stress are therefore commonly included in NM hazard assessment strategies. The fluorescence-based dichloro-dihydro-fluorescein (DCFH) assay for cellular oxidative stress is a simple and cost-effective assay, making it a good candidate assay for SSbD hazard testing strategies. It is however subject to several pitfalls and caveats. Here, we provide further optimizations to the assay using 5-(6)-Chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetyl ester (CM-H<sub>2</sub>DCFDA-AE, referred to as DCFH probe), known for its improved cell retention.</p><p>We measured the release of metabolic products of the DCFH probe from cells to supernatant, direct reactions of CM-H<sub>2</sub>DCFDA-AE with positive controls, and compared the commonly used plate reader-based DCFH assay protocol with fluorescence microscopy and flow cytometry-based protocols. After loading cells with DCFH probe, translocation of several metabolic products of the DCFH probe to the supernatant was observed in multiple cell types. Translocated DCFH products are then able to react with test substances including positive controls. Our results also indicate that intracellularly oxidized fluorescent DCF is able to translocate from cells to the supernatant. In either way, this will lead to a fluorescent supernatant, making it difficult to discriminate between intra- and extra-cellular ROS production, risking misinterpretation of possible oxidative stress when measuring fluorescence on a plate reader.</p><p>The use of flow cytometry instead of plate reader-based measurements resolved these issues, and also improved assay sensitivity. Several optimizations of the flow cytometry-based DCFH ISO standard (ISO/TS 19006:2016) were suggested, including loading cells with DCFH probe before incubation with the test materials, and applying an appropriate gating strategy including live-death staining, which was not included in the ISO standard.</p><p>In conclusion, flow cytometry- and fluorescence microscopy-based read-outs are preferred over the classical plate reader-based read-out to assess the level of intracellular oxidative stress using the cellular DCFH assay.</p></div>","PeriodicalId":18786,"journal":{"name":"NanoImpact","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2452074824000314/pdfft?md5=c9b29be2e74b40b7fe022bdcac03e48c&pid=1-s2.0-S2452074824000314-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141432330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Past progress in environmental nanoanalysis and a future trajectory for atomic mass-spectrometry methods 环境纳米分析的过去进展和原子质量谱仪方法的未来轨迹。
IF 4.7 3区 环境科学与生态学
NanoImpact Pub Date : 2024-07-01 DOI: 10.1016/j.impact.2024.100518
M.D. Montaño , A.J. Goodman , J.F. Ranville
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