Characterization of synthesized silver nanoparticles and assessment of its genotoxicity potentials using the alkaline comet assay

Flower, N. Angeline Little; Balusamy, Brabu; Revathy, M. S.; Gopalakrishnan, C.; Raja, S. V. Kasmir; Murugan, S.; Kumaravel, T. S.

doi:10.1016/j.mrgentox.2011.12.003

Cited by 57 publications

(31 citation statements)

References 25 publications

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“…Table 2 depicts the extensive and dose-dependent damage to DNA after treatment of HepG2 cells and PBMC with PVA-coated AgNps. Our results corroborate those of some investigators, who reported DNA damage in the alkaline comet assay, in PBMC treated with 50 and 100 g/mL AgNps (40-60 nm size) for 3 h (Flower et al, 2012). In another study on PBMC, monocytes were able to accumulate AgNp, triggering their activation and subsequent ROS generation (Greulich et al, 2011).…”

Section: Discussionsupporting

confidence: 92%

Poly(vinyl alcohol)-coated silver nanoparticles: Activation of neutrophils and nanotoxicology effects in human hepatocarcinoma and mononuclear cells

Paino

Zucolotto

2015

Environmental Toxicology and Pharmacology

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Section: Discussionsupporting

confidence: 92%

Poly(vinyl alcohol)-coated silver nanoparticles: Activation of neutrophils and nanotoxicology effects in human hepatocarcinoma and mononuclear cells

Paino

Zucolotto

2015

Environmental Toxicology and Pharmacology

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“…In particular, Ag NP are cytotoxic and genotoxic in various cell types including cancer cell lines [9,20] , and induce DNA damage in human peripheral blood cells [21] . Consequently, the possibility to exploit their cytotoxic activity to treat proliferative diseases is of interest.…”

Section: Discussionmentioning

confidence: 99%

Short- and long-term effects of silver nanoparticles on human microvascular endothelial cells

Castiglioni¹

2014

WJBC

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croscope. RESULTS:Ag NP are cytotoxic in a dose and time dependent fashion for HMEC. At high concentrations, Ag NP determine loss of membrane integrity as demonstrated by the increased activity of lactate dehydrogenase in the culture medium. Ag NP rapidly stimulate the formation of free radicals. However, pre-incubation with Trolox, apocynin, or N-acetyl-L-cysteine, antioxidants which have different structure and act through different mechanisms, is not sufficient to prevent cytotoxicity. Ag NP also induce DNA damage dose-dependently, as shown by comet assay. When exposed to sublethal concentrations of Ag NP for long times, the cells remain viable but are growth retarded. Interestingly, removal of Ag NP partially rescues cell growth. Also genotoxicity is reversible upon removal of Ag NP from culture medium, suggesting that no permanent modifications occur. It is noteworthy that Ag NP are cytotoxic and genotoxic also for endothelial progenitors, in particular for endothelial colony-forming cells, which participate to angiogenesis. CONCLUSION:Silver nanoparticles are cytotoxic and genotoxic for human microvascular endothelial cells and might become a useful tool to control excessive angiogenesis.© 2014 Baishideng Publishing Group Inc. All rights reserved.Key words: Endothelial cells; Nanoparticles; Genotoxicity; Cytotoxicity; Angiogenesis Core tip: We studied the sensitivity to silver nanoparticles of microvascular endothelial cells, which are responsible for tissue homeostasis and fundamental in angiogenesis. Silver nanoparticles are cytotoxic and lead to membrane leakage. Cytotoxicity is not prevented by the antioxidants Trolox, N-acetyl-L-cysteine or apocynin. Silver nanoparticles also induce DNA damage as demonstrated by comet assay. When exposed to sublethal concentrations of silver nanoparticles for long times, the cells remain viable but are growth retarded. METHODS:We cultured human microvascular endothelial cells and endothelial colony-forming cells in their corresponding growth medium. Stock solutions of Ag NP were prepared in culture medium and sonicated before use. They were added at different concentrations and for different times to culture media. The toxicity of Ag NP was investigated by measuring the reduction of yellow tetrazolium salt to dark purple formazan (MTT assay) at 575 nm. After staining with trypan blue, cell proliferation was assessed by counting viable cells. The lactate dehydrogenase leakage assay was performed on culture media by following the oxidation of NADH to NAD+ and monitoring the reaction kinetically at 340 nm. Reactive oxygen species production was quantified using 2'-7'-dichlorofluorescein diacetate. The alkaline comet assay was performed after mixing the cells with low melting-point agarose. Electrophoresis was then conducted and the samples were stained with ethidium bromide and analyzed with a fluorescence mi-

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“…In another study with the human testis embryo tumor cell line NT2 and C57BL6 mouse testis wild-type cells, silver NPs were found to induce apoptosis and necrosis in time-and dose-dependent fashions (Asare et al, 2012). Some investigators also demonstrated that silver NPs induced DNA damage in several types of cells (Flower et al, 2012;Kim et al, 2011), suggesting that the potential genotoxicity of silver NPs. The cytotoxicity and genotoxicity of silver NPs are related to dose, particle size and shape, as well as aggregation state, surface chemical characteristics and electric charge.…”

Section: Introductionmentioning

confidence: 98%

Toxic responses in rat embryonic cells to silver nanoparticles and released silver ions as analyzed via gene expression profiles and transmission electron microscopy

Shi²,

Shao

et al. 2014

Nanotoxicology

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After exposing rat embryonic cells to 20 μg/mL of silver nanoparticle (NP) suspension and their released ions for different time periods, silver nanoparticles were found in cellular nuclei, mitochondria, cytoplasm and lysosomes by transmission electron microscopy (TEM). We also observed mitochondrial destruction, distension of endoplasmic reticulum and apoptotic bodies. Global gene expression analysis showed a total of 279 genes that were up-regulated and 389 genes that were down-regulated in the silver-NP suspension exposure group, while 3 genes were up-regulated and 41 genes were down-regulated in the silver ion exposure group. Further, the GO pathway analysis suggested that these differentially expressed genes are involved in several biological processes, such as energy metabolism, oxygen transport, enzyme activities, molecular binding, etc. It is possible that inhibition of oxygen transport is mediated by the significant down-regulation of genes of the globin family, which might play an important role in silver ion-induced toxicity. KEGG pathway analysis showed that there were 23 signal pathways that were affected in the cells after exposure to silver-NP suspension, but not silver ion alone. The most significant change concerned inflammatory signal pathways, which were only found in silver-NP suspension exposed cells, indicating that inflammatory response might play an important role in the mechanism(s) of silver-NP-induced toxicity. The significant up-regulation of matrix metalloproteinases 3 and 9 suggests that silver NPs could induce extracellular matrix degradation via an inflammatory signaling pathway. The significant up-regulation of secretory leukocyte peptidase inhibitor and serine protease inhibitor 2c was considered to be an embryonic cellular defense mechanism in response to silver-NP-induced inflammation.

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Characterization of synthesized silver nanoparticles and assessment of its genotoxicity potentials using the alkaline comet assay

Cited by 57 publications

References 25 publications

Poly(vinyl alcohol)-coated silver nanoparticles: Activation of neutrophils and nanotoxicology effects in human hepatocarcinoma and mononuclear cells

Poly(vinyl alcohol)-coated silver nanoparticles: Activation of neutrophils and nanotoxicology effects in human hepatocarcinoma and mononuclear cells

Short- and long-term effects of silver nanoparticles on human microvascular endothelial cells

Toxic responses in rat embryonic cells to silver nanoparticles and released silver ions as analyzed via gene expression profiles and transmission electron microscopy

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