Genotoxicity Of Silver Nanoparticles Research Articles

Synthesis, antibacterial activity and genotoxicity of silver nanoparticles

Synthesis, antibacterial activity and genotoxicity of silver nanoparticles

In vivo toxicological assessment of silver nanoparticle in edible fish, Oreochromis mossambicus.

Silver nanoparticles are the extensively utilized among all nanoparticles due to their antibacterial and wound healing properties making them highly suitable for medical and pharmaceutical applications. The field of nanoparticle toxicity is an emerging field and the present study aims to assess the biochemical, hematological and genotoxicity in Oreochromis mossambicus exposed to different concentrations of silver nanoparticles for 7 and 14days. Silver nanoparticles were synthesized by reduction of silver nitrate using trisodium citrate and was characterized using X-ray diffraction, SEM, HRTEM and DLS. Hematological parameters like RBC, WBC, Hb, HCT and MCV and for biochemical analysis, antioxidant enzymes SOD, CAT and GPX and serum enzymes AST, ALT, ACP, ALP and LDH were analyzed. Genotoxicity was studied using comet assay. Results obtained showed decrease in erythrocytes, HCT, Hb and MCV while an increase was noted in WBC on day 7 and 14. The antioxidant enzymes SOD, CAT and GPx showed a decrease and the lipid peroxidation product MDA was elevated. The serum enzymes AST, ALT, ACP ALP and LDH showed an increased activity when compared to control. DNA damage was evident by an increase in % TDNA. The results indicate hematological, biochemical and genotoxicity of silver nanoparticles that might be mediated through ROS generation in O. mossambicus.

Biogenic Silver Nanoparticles: Assessment of Their Cytotoxicity, Genotoxicity and Study of Capping Proteins.

The development of nanotechnology in the last two decades has led to the use of silver nanoparticles (AgNPs) in various biomedical applications, including antimicrobial, anti-inflammatory, and anticancer therapies. However, the potential of the medical application of AgNPs depends on the safety of their use. In this work, we assessed the in vitro cytotoxicity and genotoxicity of silver nanoparticles and identified biomolecules covering AgNPs synthesized from actinobacterial strain SH11. The cytotoxicity of AgNPs against MCF-7 human breast cancer cell line and murine macrophage cell line RAW 264.7 was studied by MTT assay, cell LDH (lactate dehydrogenase) release, and the measurement of ROS (reactive oxygen species) level while genotoxicity in Salmonella typhimurium cells was testing using the Ames test. The in vitro analysis showed that the tested nanoparticles demonstrated dose-dependent cytotoxicity against RAW 264.6 macrophages and MCF-7 breast cancer cells. Moreover, biosynthesized AgNPs did not show a mutagenic effect of S. typhimurium. The analyses and identification of biomolecules present on the surface of silver nanoparticles showed that they were associated with proteins. The SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) analysis revealed the presence of 34 and 43 kDa protein bands. The identification of proteins performed by using LC-MS/MS (liquid chromatography with tandem mass spectrometry) demonstrated their highest homology to bacterial porins. Capping biomolecules of natural origin may be involved in the synthesis process of AgNPs or may be responsible for their stabilization. Moreover, the presence of natural proteins on the surface of bionanoparticles eliminates the postproduction steps of capping which is necessary for chemical synthesis to obtain the stable nanostructures required for application in medicine.

Nephrotoxicity and genotoxicity of silver nanoparticles in juvenile rats and possible mechanisms of action.

Because of their widespread use and potential adverse effects in young developing organism, this study focused on the nephrotoxicity and genotoxicity of chronic low-dose exposure to silver nanoparticles (AgNPs) in 32 14-day-old male Wistar rats, randomly divided into three groups receiving AgNP solution (3 mg/kg body weight) intraperitoneally for one, two, or three weeks and the untreated control group (eight animals per group). When the rats were eight weeks old, blood creatinine and urine microalbumin were tested, followed by haematoxylin and eosin (H&E) staining. Proteinuria was found in the animals treated with AgNP for three weeks, and H&E staining revealed pathological changes in the kidney sections of this group. DNA damage was detected with the alkaline comet assay in the groups treated for two and three weeks. All results indicate that chronic exposure, even at a low dose, may affect animal health. The main culprit might be increased and time-dependent reactive oxygen species (ROS) production. Highly reactive ROS could cause a major structural damage to proteins and DNA, change the expression of ion channel proteins, and trigger inflammation. The findings of our in vivo experiment raise concern about nephrotoxic and genotoxic effects of silver nanoparticles in young organisms and call for further investigation of nanoparticle properties that can be modified to minimise the risks.

Cytotoxicity and genotoxicity of silver nanoparticles in Chinese Hamster ovary cell line (CHO-K1) cells

Biomedical and pharmaceutical products comprising silver nanoparticles are attracting interest due to their potent antibacterial activities. For their safe use it has become imperative to test their cyto-genotoxic potential. In the present study the cytotoxicity and genotoxicity of three different sizes of AgNPs ranging from 15 to 22 nm and at concentrations 0.005–500 μg/ml were studied in Chinese Hamster ovary cell line (CHO-K1) cells. Cytoxicity was assessed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and for genotoxicity comet, and micronucleus assays were utilized. AgNPs were able to internalize CHO-K1 cells and cause cytotoxicity at concentrations 0.005–500 μg/ml. AgNP‐induced cyto-genotoxicity in CHO-K1 cells could be attributed to its smaller primary size. AgNP-C of size ~ 15 nm was the most potent among the three AgNPs. The genotoxic response was biphasic that increased at lower concentrations (0.005–0.025 μg/ml) and decreased at higher concentrations (0.05–0.1 μg/ml) after 24 h of exposure. Such potential in vitro genotoxic effect of AgNPs remains to be further confirmed in animal cells in vivo.

Antibacterial properties of retainers with silver nanoparticles

We read with interest the article entitled “Streptococcus mutans counts in patients wearing removable retainers with silver nanoparticles vs those wearing conventional retainers: A randomized clinical trial” in the February issue of the Journal.1Farhadian N. Usefi Mashoof R. Khanizadeh S. Ghaderi E. Farhadian M. Miresmaeili A. Streptococcus mutans counts in patients wearing removable retainers with silver nanoparticles vs those wearing conventional retainers: a randomized clinical trial.Am J Orthod Dentofacial Orthop. 2016; 149: 155-160Abstract Full Text Full Text PDF Scopus (44) Google Scholar Since we are also working in this area, we had a few questions and would like some explanations, if possible. Regarding the synthesis of silver nanoparticles:•Silver nanoparticles can be synthesized by chemical, physical, and biological methods,2Iravani S. Korbekandi H. Mirmohammadi S.V. Zolfaghari B. Synthesis of silver nanoparticles: chemical, physical and biological methods.Res Pharm Sci. 2014; 9: 385-406Google Scholar and cytotoxicity varies accordingly.3de Lima R. Seabra A.B. Durán N. Silver nanoparticles: a brief review of cytotoxicity and genotoxicity of chemically and biogenically synthesized nanoparticles.J Appl Toxicol. 2012; 32: 867-879Crossref Scopus (399) Google Scholar How did the authors synthesize their silver nanoparticles?•Was the antimicrobial resin synthesized in bulk to standardize the amounts of silver nanoparticles in retainers delivered to patients? The cytotoxcity of stored silver nanoparticles is greater as compared with freshly prepared silver nanoparticles.4Kittler S. Greulich C. Diendorf J. Koller M. Epple M. Toxicity of silver nanoparticles increases during storage because of slow dissolution under release of silver ions.Chem Mater. 2010; 22: 4548-4554Crossref Scopus (923) Google Scholar•What was the ratio of polymer to silver nanoparticles? Was the minimum inhibitory concentration (that is, the concentration at which at least 50% of the bacteria are killed) calculated? Regarding the antibacterial properties:•Which species of S mutans was present in McFarland suspensions? The antibacterial effect can vary according to the species (eg, ATCC 25175). Regarding the intervention:•After appliance delivery, were the patients given any oral hygiene instructions or directed to use any mouthwashes? Coexposure of silver nanoparticles and sodium fluoride also affects the cytotoxicity.5Inkielewicz-Stepniak I. Santos-Martinez M.J. Medina C. Radomski M.W. Pharmacological and toxicological effects of co-exposure of human gingival fibroblasts to silver nanoparticles and sodium fluoride.Int J Nanomedicine. 2014; 9: 1677-1687Google Scholar Regarding patient-related outcomes:•The authors claimed that the color of the final appliance, which could be gray to black depending on the concentration of the silver nanoparticles (shown in the Fig), was acceptable to patients. Could colorless silver nanoparticles have been used instead?•Was there any reason to carry out the T2 test after 7 weeks, since the retention period was 1 year? Perhaps the antibacterial activity could have been analyzed at regular intervals, since there could be differences in silver ions released during the long term.•Cytotoxicity to human gingival fibroblasts should have been tested at the first stage before conducting the in-vivo study because silver has been claimed to have not only cytotoxicity but also genotoxicity.6AshaRani P.V. Low Kah Mun G. Hande M.P. Valiyaveettil S. Cytotoxicity and genotoxicity of silver nanoparticles in human cells.ACS Nano. 2008; 3: 279-290Crossref Scopus (2931) Google Scholar The urinalyses and hematologic tests should have been done as mentioned in the Discussion. We acknowledge the scientific inputs of Meryam S. Rizvi, associate professor, and Jahirul Mazumder, PhD student, Department of Bioscience, Jamia Millia Islamia University, Delhi, India, for their critical review of the study by Farhadian et al. Streptococcus mutans counts in patients wearing removable retainers with silver nanoparticles vs those wearing conventional retainers: A randomized clinical trialAmerican Journal of Orthodontics and Dentofacial OrthopedicsVol. 149Issue 2PreviewThe rough surface of Hawley removable appliances provides an environment for plaque accumulation, leading to enamel demineralization. The aim of this study was to evaluate the effect of silver nanoparticles incorporated into acrylic baseplates of orthodontic retainers on Streptococcus mutans colony-forming units. Full-Text PDF Authors' responseAmerican Journal of Orthodontics and Dentofacial OrthopedicsVol. 150Issue 2PreviewWe appreciate being given the opportunity to discuss the details of our study1 and hope the following explanation will be helpful. Full-Text PDF

Immobilization of horseradish peroxidase on β-cyclodextrin-capped silver nanoparticles: Its future aspects in biosensor application

ABSTRACTThis study aimed to work out a simple and high-yield procedure for the immobilization of horseradish peroxidase on silver nanoparticle. Ultraviolet–visible (UV-vis) and Fourier-transform infrared spectroscopy and transmission electron microscopy were used to characterize silver nanoparticles. Horseradish peroxidase was immobilized on β-cyclodextrin-capped silver nanoparticles via glutaraldehyde cross-linking. Single-cell gel electrophoresis (Comet assay) was also performed to confirm the genotoxicity of silver nanoparticles. To decrease toxicity, silver nanoparticles were capped with β-cyclodextrin. A comparative stability study of soluble and immobilized enzyme preparations was investigated against pH, temperature, and chaotropic agent, urea. The results showed that the cross-linked peroxidase was significantly more stable as compared to the soluble counterpart. The immobilized enzyme exhibited stable enzyme activities after repeated uses.

Genotoxicity of meso-2,3-dimercapto succinic acid-coated silver sulfide quantum dot

Event Abstract Back to Event Genotoxicity of meso-2,3-dimercapto succinic acid-coated silver sulfide quantum dot Deniz Özkan Vardar1*, Ibrahim Hacıoğlu2, Sevtap Aydin3, Funda Yağcı4 and A. N. Basaran3 1 Hitit University Sungurlu Vocational high Scholl, Department of Health, Türkiye 2 Koç University, Department of Materials Science and Engineering, Türkiye 3 Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, Türkiye 4 Koç University, Department of Chemistry, Türkiye Nanotecnology products have been used in wide applications in chemistry, electronics, energy generation, and medicine. Despite significant interest in developing quantum dots (QDs) for biomedical applications, many researchers are convinced that QDs will never be used for the treatment of patients because of their potential toxicity. In various in vitro cell culture studies, the cytotoxic properties of some QD have been demonstrated and they have been suggested to be toxic in humans. In this study, the cytotoxic properties of Ag2S-(Meso-2,3-Dimercapto Succinic acid) nanomaterials in V79 cells (Chinese lung fibroblast cell line) were determined by MTT assay. The genotoxic effects of Ag2S-(Meso-2,3-Dimercapto Succinic acid) were evaluated by the alkaline single cell gel electrophoresis. The cells were treated with Ag2S-(Meso-2,3-Dimercapto Succinic acid) at the concentrations of 5- 2000 µg/ml. No cytotoxic effect of Ag2S-(Meso-2,3-Dimercapto Succinic acid) at all concentrations studied was observed. No significant increases in DNA damage were found at the studied concentrations when compared to negative control in V79 cells. In conclusion, further in vitro and in vivo studies are required to determine the safety doses of Ag2S-(Meso-2,3-Dimercapto Succinic acid). Acknowledgements This study was funded in part by a grant from TUBITAK (TUBITAK 3001, contract grant number: 114S861). References Ahamed, M., Karns, M., Goodson, M., Rowe, J., Hussain, S.M., Schlager, J.J., Hong, Y. 2008. “DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells”, Toxicol Appl Pharmacol., 233(3), 404-10. Ahamed, M., Siddiqui, M.A., Akhtar, M.J., Ahmad, I., Pant, A.B., Alhadlaq, H.A. 2010. “Genotoxic potential of copper oxide nanoparticles in human lung epithelial cells”, Biochem Biophys Res Commun., 396(2), 578-83. AshaRani, P.V., Low Kah Mun G., Hande, M.P., Valiyaveettil, S. 2009. “Cytotoxicity and genotoxicity of silver nanoparticles in human cells”, ACS Nano., 3(2), 279-90. Chaung, W. Mi,L.J., Boorstein, R.J. 1997. “The p53 status of Chinese hamster V79 cells frequently used for studies on DNA damage and DNA repair”, Nucleic Acids Res., 25(5), 992-4. Lunov, O., Syrovets, T., Büchele, B., Jiang, X., Röcker, C., Tron, K., Nienhaus, G.U., Walther, P., Mailänder, V., Landfester, K., Simmet, T. 2010. “The effect of carboxydextran-coated superparamagnetic iron oxide nanoparticles on c-Jun N-terminal kinase-mediated apoptosis in human macrophages” Biomaterials, 31(19), 5063-7. Park, E.J., Yi, J., Chung, K.H., Ryu, D.Y., Choi, J., Park, K. 2008. “Oxidative stress and apoptosis induced by titanium dioxide nanoparticles in cultured BEAS-2B cells”, Toxicol Lett., 180(3), 222-9. Keywords: Quantum dots, MTT assay, Comet Assay, Chinese lung fibroblast cell line, Genotoxicity, Nanotoxicity Conference: ICAW 2015 - 11th International Comet Assay Workshop, Antwerpen, Belgium, 1 Sep - 4 Sep, 2015. Presentation Type: Poster Discussion Topic: Genotoxicity testing of chemicals and nanomaterials Citation: Özkan Vardar D, Hacıoğlu I, Aydin S, Yağcı F and Basaran AN (2015). Genotoxicity of meso-2,3-dimercapto succinic acid-coated silver sulfide quantum dot. Front. Genet. Conference Abstract: ICAW 2015 - 11th International Comet Assay Workshop. doi: 10.3389/conf.fgene.2015.01.00018 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 02 Jun 2015; Published Online: 23 Jun 2015. * Correspondence: Mrs. Deniz Özkan Vardar, Hitit University Sungurlu Vocational high Scholl, Department of Health, Çorum, 19300, Türkiye, denizozkantr@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Deniz Özkan Vardar Ibrahim Hacıoğlu Sevtap Aydin Funda Yağcı A. N Basaran Google Deniz Özkan Vardar Ibrahim Hacıoğlu Sevtap Aydin Funda Yağcı A. N Basaran Google Scholar Deniz Özkan Vardar Ibrahim Hacıoğlu Sevtap Aydin Funda Yağcı A. N Basaran PubMed Deniz Özkan Vardar Ibrahim Hacıoğlu Sevtap Aydin Funda Yağcı A. N Basaran Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. 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Cytotoxicity and genotoxicity of silver nanoparticles on isolated rat hepatocytes

Cytotoxicity and genotoxicity of silver nanoparticles on isolated rat hepatocytes

Genotoxicity of Silver Nanoparticles in Lung Cells of Sprague Dawley Rats after 12 Weeks of Inhalation Exposure

Due to the widespread use of silver nanoparticles in consumer products, the toxicity of silver nanoparticles has also been studied in relation to their application. However, most genotoxicity studies of silver nanoparticles have been performed in vitro. Therefore, this study evaluated the DNA damage to lung cells caused by repeated inhalation of silver nanoparticles. Male Sprague Dawley rats were exposed to silver nanoparticles for 12 weeks in a whole-body inhalation chamber. The animals were divided into one control group and three dose groups that were exposed to silver nanoparticles (14–15 nm diameter) at concentrations of 0.66 × 106 particles/cm3 (49 μg/m3, low dose), 1.41 × 106 particles/cm3 (117 μg/m3, middle dose), and 3.24 × 106 particles /cm3 (381 μg/m3, high dose), respectively, for six hours/day over 12 weeks. The rats were sacrificed after the 12-week exposure period and the DNA damage assessed using a Comet assay of cells obtained from the right lungs. The olive tail moment values were 2.93 ± 0.19, 3.81 ± 0.23, 3.40 ± 0.22, and 5.16 ± 0.32 for the control, low-, middle-, and high-dose groups, respectively. Although no dose-dependent results were observed, a significant increase in the level of DNA damage was noted for the high-dose group.

Genotoxicity of silver nanoparticles in Vicia faba: a pilot study on the environmental monitoring of nanoparticles.

The use of silver nanoparticles (AgNPs) in commercial products has increased significantly in recent years. Although there have been some attempts to determine the toxic effects of AgNPs in mammalian and human cell-lines, there is little information on plants which play a vital role in ecosystems. The study reports the use of Vicia faba root-tip meristem to investigate the genotoxicity of AgNPs under modified GENE-TOX test conditions. The root tip cells of V. faba were treated with four different concentrations of engineered AgNPs dispersion to study toxicological endpoints such as mitotic index (MI), chromosomal aberrations (CA) and micronucleus induction (MN). For each concentration, five sets of microscopy observations were carried out. The results demonstrated that AgNPs exposure significantly increased (p < 0.05) the number of chromosomal aberrations, micronuclei, and decreased the MI in exposed groups compared to control. From this study we infer that AgNPs might have penetrated the plant system and may have impaired mitosis causing CA and MN. The results of this study demonstrate that AgNPs are genotoxic to plant cells. Since plant assays have been integrated as a genotoxicity component in risk assessment for detection of environmental mutagens, they should be given full consideration when evaluating the overall toxicological impact of the nanoparticles in the environment.

The effect of particle size on the cytotoxicity, inflammation, developmental toxicity and genotoxicity of silver nanoparticles

Silver nanoparticles are of interest to be used as antimicrobial agents in wound dressings and coatings in medical devices, but potential adverse effects have been reported in the literature. The most pronounced effect of silver nanoparticles and the role of particle size in determining these effects, also in comparison to silver ions, are largely unknown. Effects of silver nanoparticles of different sizes (20, 80, 113 nm) were compared in in vitro assays for cytotoxicity, inflammation, genotoxicity and developmental toxicity. Silver nanoparticles induced effects in all endpoints studied, but effects on cellular metabolic activity and membrane damage were most pronounced. In all toxicity endpoints studied, silver nanoparticles of 20 nm were more toxic than the larger nanoparticles. In L929 fibroblasts, but not in RAW 264.7 macrophages, 20 nm silver nanoparticles were more cytotoxic than silver ions. Collectively, these results indicate that effects of silver nanoparticles on different toxic endpoints may be the consequence of their ability to inflict cell damage. In addition, the potency of silver in the form of nanoparticles to induce cell damage compared to silver ions is cell type and size-dependent.

Differential expression of ribosomal protein gene, gonadotrophin releasing hormone gene and Balbiani ring protein gene in silver nanoparticles exposed Chironomus riparius

The eco- and genotoxicity of silver nanoparticles (AgNPs) was investigated in the fourth instar larvae of the aquatic midge, Chironomus riparius. AgNPs did not have acute toxicity in C. riparius, but did exhibited chronic toxicity on development (pupation and emergence failure) and reproduction. Genotoxicity also occurred in AgNPs exposed C. riparius. Differential Display PCR (DD-PCR), based on the Annealing Control Primer (ACP) technique, was conducted to investigate the underlying toxic mechanism, which identified altered gene expression in C. riparius after treatment with AgNPs. The possible toxicity mechanism of AgNPs in C. riparius involves the down regulation of the ribosomal protein gene (CrL15) affecting the ribosomal assembly and consequently, protein synthesis. Up regulation of the gonadotrophin releasing hormone gene (CrGnRH1) might lead to the activation of gonadotrophin releasing hormone mediated signal transduction pathways and reproductive failure. Up regulation of the Balbiani ring protein gene (CrBR2.2) may be an indication of the organism's protection mechanism against the AgNPs. The overall results suggest that the toxicity of AgNPs towards aquatic organisms should be thoroughly investigated to allow for their safe use, as they seem to exhibit important toxicity towards C. riparius.

Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549

Nanomaterials, especially silver nanoparticles (Ag NPs), are used in a rapidly increasing number of commercial products. Accordingly, the hazards associated with human exposure to nanomaterials should be investigated to facilitate the risk assessment process. A potential route of exposure to NPs is through the respiratory system. In the present study, we investigated the effects of well-characterized PVP-coated Ag NPs and silver ions (Ag+) in the human, alveolar cell line, A549. Dose-dependent cellular toxicity caused by Ag NPs and Ag+ was demonstrated by the MTT and annexin V/propidium iodide assays, and evidence of Ag NP uptake could be measured indirectly by atomic absorption spectroscopy and flow cytometry. The cytotoxicity of both silver compounds was greatly decreased by pretreatment with the antioxidant, N-acetyl-cysteine, and a strong correlation between the levels of reactive oxygen species (ROS) and mitochondrial damage (r(s) = -0.8810; p = 0.0039) or early apoptosis (r(s) = 0.8857; p = 0.0188) was observed. DNA damage induced by ROS was detected as an increase in bulky DNA adducts by (32)P postlabeling after Ag NP exposure. The level of bulky DNA adducts was strongly correlated with the cellular ROS levels (r(s) = 0.8810, p = 0.0039) and could be inhibited by antioxidant pretreatment, suggesting Ag NPs as a mediator of ROS-induced genotoxicity.