Interference of engineered nanomaterials in flow cytometry: A case study

Bohmer, Nils; Rippl, Alexandra; May, Sarah; Walter, Aurélie; Heo, Min; Kwak, Minjeong; Roesslein, Matthias; Song, Nam Woong; Wick, Peter; Hirsch, Cordula

doi:10.1016/j.colsurfb.2018.09.021

Cited by 26 publications

(29 citation statements)

References 37 publications

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“…In the absence of serum proteins, SiO 2 NPs induced false-positive signals, whereas gold NPs provoked fluorescence quenching effects. 266 For the design of NPs used in biomedical applications, it is important to obtain quantitative data about NPs uptake and at the same time information about spatial distribution of the NPs in live target cells. For this reason, imaging FC has been developed recently, which enables wide field imaging combined with high-throughput.…”

Section: Flow Cytometrymentioning

confidence: 99%

Understanding nanoparticle endocytosis to improve targeting strategies in nanomedicine

et al. 2021

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Section: Flow Cytometrymentioning

confidence: 99%

Understanding nanoparticle endocytosis to improve targeting strategies in nanomedicine

et al. 2021

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“…It is also important to note that some ENMs have been shown to cause false-positive or false-negative results in a viability assay to test for apoptosis or necrosis using flow cytometry and thus careful control experiments also need to be included for bioaccumulation measurements to avoid artifacts. 202…”

Section: Single-celled Organismsmentioning

confidence: 99%

Strategies for robust and accurate experimental approaches to quantify nanomaterial bioaccumulation across a broad range of organisms

Petersen

Mortimer

Burgess

et al. 2019

Environ. Sci.: Nano

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One of the key components for environmental risk assessment of engineered nanomaterials (ENMs) is data on bioaccumulation potential. Accurately measuring bioaccumulation can be critical for regulatory decision making regarding material hazard and risk, and for understanding the mechanism of toxicity. This perspective provides expert guidance for performing ENM bioaccumulation measurements across a broad range of test organisms and species. To accomplish this aim, we critically evaluated ENM bioaccumulation within three categories of organisms: single-celled species, multicellular species excluding plants, and multicellular plants. For aqueous exposures of suspended single-celled and small multicellular species, it is critical to perform a robust procedure to separate suspended ENMs and small organisms to avoid overestimating bioaccumulation. For many multicellular organisms, it is essential to differentiate between the ENMs adsorbed to external surfaces or in the digestive tract and the amount absorbed across epithelial tissues. For multicellular plants, key considerations include how exposure route and the role of the rhizosphere may affect the quantitative measurement of uptake, and that the efficiency of washing procedures to remove loosely attached ENMs to the roots is not well understood. Within each organism category, case studies are provided to illustrate key methodological considerations for conducting robust bioaccumulation experiments for different species within each major group. The full scope of ENM bioaccumulation measurements and interpretations are discussed including conducting the organism exposure, separating organisms from the ENMs in the test media after exposure, analytical methods to quantify ENMs in the tissues or cells, and modeling the ENM bioaccumulation results. One key finding to improve bioaccumulation measurements was the critical need for further analytical method development to identify and quantify ENMs in complex matrices. Overall, the discussion, suggestions, and case studies described herein will help improve the robustness of ENM bioaccumulation studies.

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“…Detailed material characterization of Au-NPs has been described by Bohmer et al 36 Transmission electron microscopy (TEM) analysis was used to characterize the primary particle size and morphology of the three different Au-NPs. Analysis of TEM images revealed particles with a diameter of 3.1 ± 1.3 nm for Au-NP I, 2.6 ± 1.2 nm for Au-NP II and 2.2 ± 0.8 nm for Au-NP III.…”

Section: Physico-chemical Characterization Of Au-npsmentioning

confidence: 99%

Transient DNA damage following exposure to gold nanoparticles

et al. 2018

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Due to their interesting physicochemical properties, gold nanoparticles (Au-NPs) are the focus of increasing attention in the field of biomedicine and are under consideration for use in drug delivery and bioimaging, or as radiosensitizers and nano-based vaccines. Thorough evaluation of the genotoxic potential of Au-NPs is required, since damage to the genome can remain undetected in standard hazard assessments. Available genotoxicity data is either limited or contradictory. Here, we examined the influence of three surface modified 3-4 nm Au-NPs on human A549 cells, according to the reactive oxygen species (ROS) paradigm. After 24 h of Au-NP treatment, nanoparticles were taken up by cells as agglomerates; however, no influence on cell viability or inflammation was detected. No increase in ROS production was observed by H2-DCF assay; however, intracellular glutathione levels reduced over time, indicating oxidative stress. All three types of Au-NPs induced DNA damage, as detected by alkaline comet assay. The strongest genotoxic effect was observed for positively charged Au-NP I. Further analysis of Au-NP I by neutral comet assay, fluorimetric detection of alkaline DNA unwinding assay, and γH2AX staining, revealed that the induced DNA lesions were predominantly alkali-labile sites. As highly controlled repair mechanisms have evolved to remove a wide range of DNA lesions with great efficiency, it is important to focus on both acute cyto- and genotoxicity, alongside post-treatment effects and DNA repair. We demonstrate that Au-NP-induced DNA damage is largely repaired over time, indicating that the observed damage is of transient nature.

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Interference of engineered nanomaterials in flow cytometry: A case study

Cited by 26 publications

References 37 publications

Understanding nanoparticle endocytosis to improve targeting strategies in nanomedicine

Understanding nanoparticle endocytosis to improve targeting strategies in nanomedicine

Strategies for robust and accurate experimental approaches to quantify nanomaterial bioaccumulation across a broad range of organisms

Transient DNA damage following exposure to gold nanoparticles

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