Critical factors to be considered when testing nanomaterials for genotoxicity with the comet assay

Huk, Anna; Collins, Andrew; Yamani, Naouale El; Porredon, C.; Azqueta, Amaya; Lapuente, Joaquín de; Dušinská, Maria

doi:10.1093/mutage/geu077

Cited by 38 publications

(31 citation statements)

References 9 publications

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Section: Dna Damagementioning

confidence: 99%

“…42 It should also be emphasized that no modifications of the assay are necessary during the testing of NPs. 42 In our experiments, positive and negative controls were included to demonstrate correct performance of the assay and ensure reproducibility. Untreated cells were used as negative controls to show the background level of DNA damage, whereas the cells treated with hydrogen peroxide, which induces strand breaks, were used as positive controls.…”

Section: Dna Damagementioning

confidence: 99%

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Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles

Pongrac¹,

Pavičić²,

Milić³

et al. 2016

IJN

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Biocompatibility, safety, and risk assessments of superparamagnetic iron oxide nanoparticles (SPIONs) are of the highest priority in researching their application in biomedicine. One improvement in the biological properties of SPIONs may be achieved by different functionalization and surface modifications. This study aims to investigate how a different surface functionalization of SPIONs – uncoated, coated with d -mannose, or coated with poly- l -lysine – affects biocompatibility. We sought to investigate murine neural stem cells (NSCs) as important model system for regenerative medicine. To reveal the possible mechanism of toxicity of SPIONs on NSCs, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, cell-membrane potential, DNA damage, and activities of SOD and GPx were examined. Even in cases where reactive oxygen species levels were significantly lowered in NSCs exposed to SPIONs, we found depleted intracellular glutathione levels, altered activities of SOD and GPx, hyperpolarization of the mitochondrial membrane, dissipated cell-membrane potential, and increased DNA damage, irrespective of the surface coating applied for SPION stabilization. Although surface coating should prevent the toxic effects of SPIONs, our results showed that all of the tested SPION types affected the NSCs similarly, indicating that mitochondrial homeostasis is their major cellular target. Despite the claimed biomedical benefits of SPIONs, the refined determination of their effects on various cellular functions presented in this work highlights the need for further safety evaluations. This investigation helps to fill the knowledge gaps on the criteria that should be considered in evaluating the biocompatibility and safety of novel nanoparticles.

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Section: Dna Damagementioning

confidence: 99%

Section: Dna Damagementioning

confidence: 99%

Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles

Pongrac¹,

Pavičić²,

Milić³

et al. 2016

IJN

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“…Cos‐1 fibroblast‐like kidney cells were treated with different concentrations of iron oxide NMs, and cells embedded in minigels (12 per slide) Subsequent incubation with FPG revealed damage not seen with the basic assay for strand breaks (without FPG) 71. Further, Huk et al used the 12 gel system to study genotoxicity of nine well‐defined nanosilvers in relation to their size and surface properties 126, 127. Reservations have been expressed about the use of the comet assay, because of potential interference of NMs with the assay.…”

Section: High‐throughput Comet Assaymentioning

confidence: 99%

High throughput toxicity screening and intracellular detection of nanomaterials

Collins

Annangi

Rubio

et al. 2016

WIREs Nanomed Nanobiotechnol

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116

106

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With the growing numbers of nanomaterials (NMs), there is a great demand for rapid and reliable ways of testing NM safety—preferably using in vitro approaches, to avoid the ethical dilemmas associated with animal research. Data are needed for developing intelligent testing strategies for risk assessment of NMs, based on grouping and read‐across approaches. The adoption of high throughput screening (HTS) and high content analysis (HCA) for NM toxicity testing allows the testing of numerous materials at different concentrations and on different types of cells, reduces the effect of inter‐experimental variation, and makes substantial savings in time and cost. HTS/HCA approaches facilitate the classification of key biological indicators of NM‐cell interactions. Validation of in vitro HTS tests is required, taking account of relevance to in vivo results. HTS/HCA approaches are needed to assess dose‐ and time‐dependent toxicity, allowing prediction of in vivo adverse effects. Several HTS/HCA methods are being validated and applied for NM testing in the FP7 project NANoREG, including Label‐free cellular screening of NM uptake, HCA, High throughput flow cytometry, Impedance‐based monitoring, Multiplex analysis of secreted products, and genotoxicity methods—namely High throughput comet assay, High throughput in vitro micronucleus assay, and γH2AX assay. There are several technical challenges with HTS/HCA for NM testing, as toxicity screening needs to be coupled with characterization of NMs in exposure medium prior to the test; possible interference of NMs with HTS/HCA techniques is another concern. Advantages and challenges of HTS/HCA approaches in NM safety are discussed. WIREs Nanomed Nanobiotechnol 2017, 9:e1413. doi: 10.1002/wnan.1413For further resources related to this article, please visit the WIREs website.

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“…The first one deals with nanoparticles and comprises variation of size of the nanoparticles, variations of size of distribution, various purities of the nanomaterials with the same average size of nanoparticles, differences of their coatings, differences of crystal structure of the types of nanomaterials, differences of size of aggregates in solution or medium, and different concentrations of nanomaterials used in assay test. The second group comprises testing conditions such as cell number, cell culture plate format and volume of treatment medium on the plate, and differences in assays [42,43].…”

Section: Genotoxicitymentioning

confidence: 99%

Medical Aspects of Nanomaterial Toxicity

Siemianowicz¹,

Likus²,

Markowski³

2015

Nanomaterials - Toxicity and Risk Assessment

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Nanosilver is the most popular and most studied nanomaterial, however, a family of nanomaterials is rapidly enlarging. They are used in various branches of industry and everyday life. In medicine new nanomaterials can be used either alone or in combination with other "classical" drugs, e.g. cytostatic drugs or antibiotics. They can be also used as diagnostic agents. A development of nanoparticles has led to a new combination of diagnostic and therapy -theranostic. Size of a particle makes a difference not only between bulk material and nanomaterial, but also in their properties and toxicity. Nanomaterials can have beneficial properties, but can also be toxic. New issues concerning nanomaterials arise -an industrial exposure and environmental pollution. They can enter human body in various ways. Cellular mechanisms of nanomaterial toxicity comprise mainly a generation of reactive oxygen species and genotoxicity. The differences between toxicity of fine particles and nanoparticles have led to an origin of a new branch of science, nanotoxicology.

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Critical factors to be considered when testing nanomaterials for genotoxicity with the comet assay

Cited by 38 publications

References 9 publications

Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles

Oxidative stress response in neural stem cells exposed to different superparamagnetic iron oxide nanoparticles

High throughput toxicity screening and intracellular detection of nanomaterials

Medical Aspects of Nanomaterial Toxicity

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