Interlaboratory Evaluation of
            <i>in Vitro</i>
            Cytotoxicity and Inflammatory Responses to Engineered Nanomaterials: The NIEHS Nano GO Consortium

Xia, Tian; Hamilton, Raymond F.; Bonner, James C.; Crandall, Edward D.; Elder, Alison; Fazlollahi, Farnoosh; Girtsman, Teri; Kim, Kwang Jin; Mitra, Somenath; Ntim, Susana Addo; Orr, Galya; Tagmount, Mani; Taylor, Alexia J.; Telesca, Donatello; Tolic, Ana; Vulpe, Chris D.; Walker, Andrea J.; Wang, Xiang; Witzmann, Frank A.; Wu, Nianqiang; Xie, Yumei; Zink, Jeffery I.; Nel, André E.; Holian, Andrij

doi:10.1289/ehp.1306561

Cited by 183 publications

(203 citation statements)

References 26 publications

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“…This issue was also observed in previous ENM interlaboratory comparison using the MTS assay. Xia et al added a centrifugation step to their protocol to remove NP interference effects (Xia et al, 2013). Testing the impact of a centrifugation step or modified rinsing steps to decrease ENM interference will be important for optimizing the MTS protocol described here for better harmonization between laboratories.…”

Section: Discussionmentioning

confidence: 99%

“…This study differs from a recently published interlaboratory comparison (Xia et al, 2013) in that we systematically evaluate the contributions to the total variability from the various steps of the assay and provide numerous specifications that ensure comparability of the assay measurement process. This study was not designed as a comparison of the effect of different cell lines or various ENMs; the objective was to show the power of using an assay design with system control measurements in combination with an interlaboratory comparison experiment to systematically understand the sources of variability in the assay.…”

Section: Introductionmentioning

confidence: 99%

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Toward achieving harmonization in a nanocytotoxicity assay measurement through an interlaboratory comparison study

Elliott

Rösslein

Song

et al. 2017

ALTEX

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SummaryDevelopment of reliable cell-based nanotoxicology assays is important for evaluation of potentially hazardous engineered nanomaterials. Challenges to producing a reliable assay protocol include working with nanoparticle dispersions and living cell lines, and the potential for nano-related interference effects. Here we demonstrate the use of a 96-well plate design with several measurement controls and an interlaboratory comparison study involving five laboratories to characterize the robustness of a nanocytotoxicity MTS cell viability assay based on the A549 cell line. The consensus EC 50 values were 22.1 mg/L (95% confidence intervals 16.9 mg/L to 27.2 mg/L) and 52.6 mg/L (44.1 mg/L to 62.6 mg/L) for positively charged polystyrene nanoparticles for the serum-free and serum conditions, respectively, and 49.7 µmol/L (47.5 µmol/L to 51.5 µmol/L) and 77.0 µmol/L (54.3 µmol/L to 99.4 µmol/L) for positive chemical control cadmium sulfate for the serum-free and serum conditions, respectively. Results from the measurement controls can be used to evaluate the sources of variability and their relative magnitudes within and between laboratories. This information revealed steps of the protocol that may need to be modified to improve the overall robustness and precision. The results suggest that protocol details such as cell line ID, media exchange, cell handling, and nanoparticle dispersion are critical to ensure protocol robustness and comparability of nanocytotoxicity assay results. The combination of system control measurements and interlaboratory comparison data yielded insights that would not have been available by either approach by itself.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Toward achieving harmonization in a nanocytotoxicity assay measurement through an interlaboratory comparison study

Elliott

Rösslein

Song

et al. 2017

ALTEX

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“…Above all, CNT doses associated with genotoxicity should be determined by in vitro and in vivo studies. To avoid differences in interlaboratory research protocols contributing to conflicting data in the literature, consortium research programmes are emerging to reduce variability and validate findings [165]. In parallel, a number of workplaces should be monitored, from research laboratories to CNT manufacturing, manipulating and recycling plants.…”

Section: Potential Mechanisms Of Cnt Oncogenesismentioning

confidence: 99%

Malignant pleural mesothelioma: history, controversy and future of a manmade epidemic

2015

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Asbestos is the term for a family of naturally occurring minerals that have been used on a small scale since ancient times. Industrialisation demanded increased mining and refining in the 20th century, and in 1960, Wagner, Sleggs and Marchand from South Africa linked asbestos to mesothelioma, paving the way to the current knowledge of the aetiology, epidemiology and biology of malignant pleural mesothelioma. Pleural mesothelioma is one of the most lethal cancers, with increasing incidence worldwide. This review will give some snapshots of the history of pleural mesothelioma discovery, and the body of epidemiological and biological research, including some of the controversies and unresolved questions. Translational research is currently unravelling novel circulating biomarkers for earlier diagnosis and novel treatment targets. Current breakthrough discoveries of clinically promising noninvasive biomarkers, such as the 13-protein signature, microRNAs and the BAP1 mesothelioma/cancer syndrome, are highlighted. The asbestos history is a lesson to not be repeated, but here we also review recent in vivo and in vitro studies showing that manmade carbon nanofibres could pose a similar danger to human health. This should be taken seriously by regulatory bodies to ensure thorough testing of novel materials before release in the society. @ERSpublications Malignant pleural mesothelioma is a cancer with increasing death tolls due to the past and present use of asbestos http://ow.ly/DhA2y

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“…The concentration range evaluated for cytotoxicity was selected based on previous in vitro experimentation of other NPs. 22 Changes in cell viability were assessed using the MTS assay (Promega Corporation, Fitchburg, WI, USA) via manufacturer's instructions using a spectrophotometer (BioTek Synergy HT; BioTek, Winooski, VT, USA).…”

Section: Evaluation Of Agnp Cytotoxicitymentioning

confidence: 99%

A hyperspectral and toxicological analysis of protein corona impact on silver nanoparticle properties, intracellular modifications, and macrophage activation

Shannahan¹,

Podila²,

Brown³

2015

IJN

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The inevitable adsorption of biomolecules on nanomaterials results in the formation of a protein corona (PC), which modifies the nanoparticle (NP)–cell interface resulting in modified uptake, activity, clearance, and toxicity. While the physicochemical properties of the NP govern the composition of PC, the formation of PC in turn alters the characteristics of the NP by imparting a new unique “biological” identity. To assess how the PC influences AgNP properties, intracellular modifications, and cellular responses, we utilized a combination of hyperspectral and toxicological analyses. AgNPs were coated with a complex PC (multiple proteins, eg, 10% fetal bovine serum) or a simple PC (single protein, eg, bovine serum albumin [BSA]) and evaluated by hyperspectral and dynamic light scattering for modifications in AgNP properties. Mouse macrophages were exposed to AgNPs with PCs and examined for differences in uptake, cytotoxicity, and cell activation. Hyperspectral imaging revealed intracellular modifications to AgNPs that were found to spectrally match alterations in AgNPs following incubation in lysosomal fluid. Addition of the PC influenced AgNP uptake and cytotoxicity; however, hydrodynamic size and surface charge did not contribute to these responses. Assessments of all endpoints demonstrated differences between complex and BSA PC, suggesting that these responses are not purely driven by the primary protein component of the complex PC (ie, BSA). Alterations in cellular–NP uptake/interactions may be driven through cell surface receptor recognition of protein constituents that make up the PC rather than the physicochemical differences in AgNPs.

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Interlaboratory Evaluation of in Vitro Cytotoxicity and Inflammatory Responses to Engineered Nanomaterials: The NIEHS Nano GO Consortium

Cited by 183 publications

References 26 publications

Toward achieving harmonization in a nanocytotoxicity assay measurement through an interlaboratory comparison study

Toward achieving harmonization in a nanocytotoxicity assay measurement through an interlaboratory comparison study

Malignant pleural mesothelioma: history, controversy and future of a manmade epidemic

A hyperspectral and toxicological analysis of protein corona impact on silver nanoparticle properties, intracellular modifications, and macrophage activation

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