Genotoxicity Of AgNPs Research Articles

Synergistic antibacterial mechanism of silver-copper bimetallic nanoparticles.

The excessive use of antibiotics in clinical settings has resulted in the rapid expansion, evolution, and development of bacterial and microorganism resistance. It causes a significant challenge to the medical community. Therefore, it is important to develop new antibacterial materials that could replace traditional antibiotics. With the advancements in nanotechnology, it has become evident that metallic and metal oxide nanoparticles (MeO NPs) exhibit stronger antibacterial properties than their bulk and micron-sized counterparts. The antibacterial properties of silver nanoparticles (Ag NPs) and copper nanoparticles (Cu NPs) have been extensively studied, including the release of metal ions, oxidative stress responses, damages to cell integrity, and immunostimulatory effects. However, it is crucial to consider the potential cytotoxicity and genotoxicity of Ag NPs and Cu NPs. Numerous experimental studies have demonstrated that bimetallic nanoparticles (BNPs) composed of Ag NPs and Cu NPs exhibit strong antibacterial effects while maintaining low cytotoxicity. Bimetallic nanoparticles offer an effective means to mitigate the genotoxicity associated with individual nanoparticles while considerably enhancing their antibacterial efficacy. In this paper, we presented on various synthesis methods for Ag-Cu NPs, emphasizing their synergistic effects, processes of reactive oxygen species (ROS) generation, photocatalytic properties, antibacterial mechanisms, and the factors influencing their performance. These materials have the potential to enhance efficacy, reduce toxicity, and find broader applications in combating antibiotic resistance while promoting public health.

Green synthesized silver nanoparticles induced cytogenotoxic and genotoxic changes in Allium cepa L. varies with nanoparticles doses and duration of exposure.

This study aimed to test the ability of aqueous leaf extract of Eichhornia crassipes (Mart.) Solms-Laubach to synthesize silver nanoparticles (AgNPs) and to estimate the cytotoxicity and genotoxicity of AgNPs using Allium cepa assay. Fresh Eichhornia crassipes plants were collected from the Nile River of Egypt. The mixed-shaped structures of the biogenic AgNPs were qualitatively characterized by UV-vis spectroscopy, scanning electron microscopy and transmission electron microscopy. Selected area electron diffraction confirmed the crystalline structure of AgNPs and energy dispersive X-ray analysis clarified the presence of the elemental silver in a percentage of 83.29%. The biogenic AgNPs were quite stable (0.316) and negatively charged (-18.5mV) based on the polydispersity index values. Allium cepa L. roots were exposed to several AgNPs concentrations (0, 5, 10, 20, 40 and 80mgL-1) for different time intervals 2, 4 and 6h. Cytotoxicity measured by both the spectrophotometric and macroscopic techniques recorded the maximum cell death of root tips of A.cepa after 20mgL-1 treatment. The analysis of comet assay output images showed an alteration of DNA repair kinetics. The use of aqueous leaf extract of E.crassipes (Mart.) Solms-Laubach in the large-scale production of AgNPs by the method proposed in this study may be a step in improving the water loss in the Nile River. At the same time, a sensitive approach to the cytogenotoxicity of AgNPs must be considered.

Antitumor activity of silver nanoparticles in Ehrlich carcinoma-bearing mice.

Silver nanoparticles (AgNPs) have a wide range of industrial and biomedical applications. The aim of the present study was to determine the cytotoxic and genotoxic effects of AgNPs on Ehrlich carcinoma-bearing mice. AgNPs were characterized by ultraviolet-visible absorption spectroscopy, dynamic light scattering, and transmission electron microscopy (TEM). Furthermore, the cytotoxicity and genotoxicity of AgNPs were evaluated using a series of assays: superoxide dismutase (SOD) enzyme activity, malondialdehyde (MDA) levels, DNA damage (comet assay), and histopathological examination of tissues and tumor size in Ehrlich carcinoma-bearing mice. Treatment of Ehrlich carcinoma-bearing mice with various concentrations of AgNPs (6, 24, and 48mg/kg) injected intra peritoneal (IP) and intra tumor (IT) revealed that AgNPs significantly elevated the levels (0.5- to 5-fold) of MDA and reduced the activity (32-64%) of SOD. Furthermore, AgNPs caused a 2- to 3-fold increase in comet parameters such as percent tail DNA. Additionally, AgNPs inhibit the promotion of Ehrlich carcinoma by masses of necrotic and fragmented tumor cells. Consequently, the volume of tumor reduced by about 31-95% compared to control one. The results indicate that AgNPs possess cytotoxic and genotoxic effects against Ehrlich tumor and confirm the antitumor properties of AgNPs.

Cytotoxicity and genotoxicity of nanosilver in stable GADD45α promoter‐driven luciferase reporter HepG2 and A549 cells

The intense commercial application of silver nanoparticles (AgNPs) has been raising concerns about their potential adverse health effects to human. This study aimed to explore the potency of AgNPs to induce GADD45α gene, an important stress sensor, and its relationships with the cytotoxicity and genotoxicity elicited by AgNPs. Two established HepG2 and A549 cell lines containing the GADD45α promoter-driven luciferase reporter were treated with increasing concentrations of AgNPs for 48 hours. After the treatment, transcriptional activation of GADD45α indicated by luciferase activity, cell viability, cell cycle arrest, and levels of genotoxicity were determined. The uptake and intracellular localization of AgNPs, cellular Ag doses as well as Ag+ release were also detected. AgNPs could activate GADD45α gene at the transcriptional level as demonstrated by the dose-dependent increases in luciferase activity in both the reporter cells. The relative luciferase activity was greater than 12× the control level in HepG2-luciferase cells at the highest concentration tested where the cell viability decreased to 17.0% of the control. These results was generally in accordance with the positive responses in cytotoxicity, cell cycle arrest of Sub G1 and G2/M phase, Olive tail moment, micronuclei frequency, and the cellular Ag content. The cytotoxicity and genotoxicity of AgNPs seems to occur mainly via particles uptake and the subsequent liberation of ions inside the cells. And furthermore, the GADD45α promoter-driven luciferase reporter cells, especially the HepG2-luciferase cells, could provide a new and valuable tool for predicting nanomaterials genotoxicity in humans.

Particle coatings but not silver ions mediate genotoxicity of ingested silver nanoparticles in a mouse model

Incorporation of silver nanoparticles (AgNPs) in toothpaste, food containers, dietary supplements and other consumer products can result in oral exposure to AgNPs and/or silver ions (Ag+) released from the surface of AgNPs. To examine whether ingestion of AgNPs or Ag+ results in genotoxic damage and whether AgNP coatings modulate the effect, we exposed mice orally to 20nm citrate-coated AgNPs, polyvinylpyrrolidone (PVP)-coated AgNPs, silver acetate or respective vehicles at a 4mg/kg dose (equivalent to 800× the EPA reference dose for Ag) for 7days. Genotoxicity was examined in the systemic circulation and bone marrow at 1, 7, and 14days post-exposure. We found that citrate-coated AgNPs induced chromosomal damage in bone marrow and oxidative DNA damage and double strand breaks in peripheral blood. These damages persisted for at least 14days after exposure termination. Because oxidative DNA damage and strand breaks are repaired rapidly, their presence after exposure cessation indicates that citrate-coated AgNPs persist in the body. In contrast, PVP-coated AgNPs and silver acetate did not induce DNA or chromosomal damage at any time point measured. To determine whether coating-dependent genotoxicity is related to different AgNP changes in the gastrointestinal tract, we examined AgNP behavior and fate in an in vitro gastrointestinal digestion model using UV–visible spectroscopy and DLS. Citrate-coated AgNPs were more susceptible to agglomeration than PVP-coated AgNPs in digestive juices with or without proteins. In summary, AgNPs but not Ag+ are genotoxic following oral ingestion. Nanoparticle coatings modulate gastrointestinal transformation and genotoxicity of AgNPs, where higher agglomeration of AgNPs in gastrointestinal juices is associated with higher genotoxicity in tissues. Since genotoxicity is a strong indicator of cancer risk, further long-term studies focusing on cancer are warranted.

Factors affecting the in vitro micronucleus assay for evaluation of nanomaterials.

A number of in vitro methodologies have been used to assess the genotoxicity of different nanomaterials, including titanium dioxide nanoparticles (TiO2 NPs) and silver nanoparticles (AgNPs). The in vitro micronucleus assay is one of the most commonly used test methods for genotoxicity evaluation of nanomaterials. However, due to the novel features of nanomaterials, such as high adsorption capacity and fluorescence properties, there are unexpected interactions with experimental components and detection systems. In this study, we evaluate the interference by two nanoparticles, AgNPs and TiO2 NPs, with the in vitro micronucleus assay system and possible confounding factors affecting cytotoxicity and genotoxicity assessment of the nanomaterials including cell lines with different p53 status, nanoparticle coatings and fluorescence, cytochalasin B, fetal bovine serum in cell treatment medium and different measurement methodologies for detecting micronuclei. Our results showed that micronucleus induction by AgNPs was similar when evaluated using flow cytometry or microscope, whereas the induction by TiO2 NPs was different using the two methods due to TiO2's fluorescence interference with the cytometry equipment. Cells with the mutated p53 gene were more sensitive to micronucleus induction by AgNPs than the p53 wild-type cells. The presence of serum during treatment increased the toxicity of AgNPs. The coatings of nanoparticles played an important role in the genotoxicity of AgNPs. These collective data highlight the importance of considering the unique properties of nanoparticles in assessing their genotoxicity using the in vitro micronucleus assay.

In Vitro Effect of 30 nm Silver Nanoparticles on Adipogenic Differentiation of Human Mesenchymal Stem Cells.

With the combined use of silver nanoparticles (Ag NPs) and human bone marrow derived mesenchymal stem cells (hMSCs) in bone tissue engineering, more knowledge of the effects of Ag NPs on hMSCs is required. Up to date, researches mainly focused on the cytotoxicity and genotoxicity of Ag NPs, only few studies discussed their influence on the differentiation of stem cells, especially adipogenic differentiation. In the present study, we investigated the in vitro uptake of 30 nm PVP-coated Ag NPs in hMSCs and their effects on cell viability, cell morphology and adipogenic differentiation of hMSCs. HMSCs were exposed to Ag NPs at concentrations of 25 and 50 μg/mL for 24 hours and at concentrations of 5 and 10 μg/mL throughout the whole differentiation period. Results of cell viability showed that Ag NPs caused time- and dose-dependent toxicity in hMSCs. Transmission electron microscopy (TEM) confirmed the uptake of Ag NPs into cytoplasm of hMSCs. No influence on cell morphology was observed. The 30 nm sized Ag NPs had no effects on adiponectin secretion, lipid droplet formation and the expression of adipogenic marker genes. It is concluded that under our experimental conditions, 30 nm PVP-coated Ag NPs do not influence the adipogenic differentiation of hMSCs in vitro. The present results provide a reference for the usage of 30 nm Ag NPs in the presence of hMSCs in bone tissue engineering.

Silver nanoparticles: correlating nanoparticle size and cellular uptake with genotoxicity.

The focus of this research was to develop a better understanding of the pertinent physico-chemical properties of silver nanoparticles (AgNPs) that affect genotoxicity, specifically how cellular uptake influences a genotoxic cell response. The genotoxicity of AgNPs was assessed for three potential mechanisms: mutagenicity, clastogenicity and DNA strand-break-based DNA damage. Mutagenicity (reverse mutation assay) was assessed in five bacterial strains of Salmonella typhimurium and Echerichia coli, including TA102 that is sensitive to oxidative DNA damage. AgNPs of all sizes tested (10, 20, 50 and 100nm), along with silver nitrate (AgNO3), were negative for mutagenicity in bacteria. No AgNPs could be identified within the bacteria cells using transmission electron microscopy (TEM), indicating these bacteria lack the ability to actively uptake AgNPs 10nm or larger. Clastogenicity (flow cytometry-based micronucleus assay) and intermediate DNA damage (DNA strand breaks as measured in the Comet assay) were assessed in two mammalian white blood cell lines: Jurkat Clone E6-1 and THP-1. It was observed that micronucleus and Comet assay end points were inversely correlated with AgNP size, with smaller NPs inducing a more genotoxic response. TEM results indicated that AgNPs were confined within intracellular vesicles of mammalian cells and did not penetrate the nucleus. The genotoxicity test results and the effect of AgNO3 controls suggest that silver ions may be the primary, and perhaps only, cause of genotoxicity. Furthermore, since AgNO3 was not mutagenic in the gram-negative bacterial Ames strains tested, the lack of bacterial uptake of the AgNPs may not be the major reason for the lack of genotoxicity observed.

Influence of silver nanoparticles on osteogenic differentiation of human mesenchymal stem cells.

Silver nanoparticles (Ag NPs) have been widely used in bone grafts due to their antimicrobial/antiviral properties. Despite extensive research focused on the cytotoxicity and genotoxicity of Ag NPs, little is known of their effect on cell differentiation. In this study, the uptake of Ag NPs and its effect on the osteogenic differentiation of human mensenchymal stem cells (hMSCs) were investigated in vitro. Ag NPs with a diameter of -30 nm were prepared. The hMSCs were exposed to different concentrations of Ag NPs, i.e., 0, 10, 50 and 100 microg/mL. The results show that Ag NPs cause cytotoxicity of hMSCs. It is also demonstrated that the uptake of Ag NPs does not affect the alkaline phosphatase activity, osteocalcin gene expression, osteopontin expression and mineralization level of hMSCs. The uptake of Ag NPs does not influence the osteogenic differentiation of hMSCs. This will give a positive reference for the application of Ag NPs in bone tissue engineering, especially their incorporation with stem cells in bone grafts.

Sensitivity of freshwater pulmonate snail Lymnaea luteola L., to silver nanoparticles

Toxicity of nanoparticles depends on many factors including size, shape, chemical composition, surface area and surface charge. Silver nanoparticles (AgNPs) are likely to enter the aquatic ecosystems because of their multiple applications and pose a health concern for humans and aquatic species. Therefore, we used a freshwater snail Lymnaea luteola L (L. luteola) to investigate the acute toxicity and genotoxicity of AgNPs in a static-renewal system for 96h. AgNPs caused molluscicidal activity in L. luteola, with 96-h median lethal concentrations (LC50) (48.10μgL−1). We have observed that AgNPs (36μgL−1) elicited a significant (p<0.01) reduction in glutathione, glutathione-s-transferase and glutathione peroxidase with a concomitant increase in malondialdehyde level and catalase in digestive gland of L. luteola. However, a significant (p<0.01) induction in DNA damage was observed by the alkaline single cell gel electrophoresis in digestive gland cells treated with AgNPs for 24 and 96h. These results demonstrate that silver nanoparticles are lethal to freshwater snail L. luteola. The oxidative stress biomarkers and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.

Appropriate &amp;lt;i&amp;gt;In Vitro&amp;lt;/i&amp;gt; Methods for Genotoxicity Testing of Silver Nanoparticles

ObjectivesWe investigated the genotoxic effects of 40-59 nm silver nanoparticles (Ag-NPs) by bacterial reverse mutation assay (Ames test), in vitro comet assay and micronucleus (MN) assay. In particular, we directly compared the effect of cytochalasin B (cytoB) and rat liver homogenate (S9 mix) in the formation of MN by Ag-NPs.MethodsBefore testing, we confirmed that Ag-NPs were completely dispersed in the experimental medium by sonication (three times in 1 minute) and filtration (0.2 µm pore size filter), and then we measured their size in a zeta potential analyzer. After that the genotoxicity were measured and especially, S9 mix and with and without cytoB were compared one another in MN assay.ResultsAmes test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains revealed that Ag-NPs with or without S9 mix did not display a mutagenic effect. The genotoxicity of Ag-NPs was also evaluated in a mammalian cell system using Chinese hamster ovary cells. The results revealed that Ag-NPs stimulated DNA breakage and MN formation with or without S9 mix in a dose-dependent manner (from 0.01 µg/mL to 10 µg/mL). In particular, MN induction was affected by cytoB.ConclusionsAll of our findings, with the exception of the Ames test results, indicate that Ag-NPs show genotoxic effects in mammalian cell system. In addition, present study suggests the potential error due to use of cytoB in genotoxic test of nanoparticles.

Genotoxic effects of silver nanoparticles stimulated by oxidative stress in human normal bronchial epithelial (BEAS-2B) cells

Many classes of silver nanoparticles (Ag-NPs) have been synthesized and widely applied, but the genotoxicity of Ag-NPs and the factors leading to genotoxicity remain unknown. Therefore, the purpose of this study is to elucidate the genotoxic effects of Ag-NPs in lung and the role of oxidative stress on the genotoxic effects of Ag-NPs. For this, Ag-NPs were completely dispersed in medium by sonication and filtration. The Ag-NPs dispersed in medium were 43–260nm in size. We observed distinct uptake of Ag-NPs into BEAS-2B cells. The Ag-NPs aggregates were wrapped with an endocytic vesicle within the cytoplasm and nucleus of BEAS-2B cells. In the comet assay and micronucleus (MN) assay for BEAS-2B cells, Ag-NPs stimulated DNA breakage and MN formation in a dose-dependent manner. The genotoxic effect of Ag-NPs was partially blocked by scavengers. In particular, of the scavengers tested, superoxide dismutase most significantly blocked the genotoxic effects in both the cytokinesis-block MN assay and the comet assay. In the modified comet assay, Ag-NPs induced a significant increase in oxidative DNA damage. Furthermore, in the oxidative stress assay, Ag-NPs significantly increased the reactive oxygen radicals. These results suggest that Ag-NPs have genotoxic effects in BEAS-2B cells and that oxidative stress stimulated by Ag-NPs may be an important factor in their genotoxic effects.

DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells

Silver nanoparticles (Ag NPs) have recently received much attention for their possible applications in biotechnology and life sciences. Ag NPs are of interest to defense and engineering programs for new material applications as well as for commercial purposes as an antimicrobial. However, little is known about the genotoxicity of Ag NPs following exposure to mammalian cells. This study was undertaken to examine the DNA damage response to polysaccharide surface functionalized (coated) and non-functionalized (uncoated) Ag NPs in two types of mammalian cells; mouse embryonic stem (mES) cells and mouse embryonic fibroblasts (MEF). Both types of Ag NPs up-regulated the cell cycle checkpoint protein p53 and DNA damage repair proteins Rad51 and phosphorylated-H2AX expression. Furthermore both of them induced cell death as measured by the annexin V protein expression and MTT assay. Our observations also suggested that the different surface chemistry of Ag NPs induce different DNA damage response: coated Ag NPs exhibited more severe damage than uncoated Ag NPs. The results suggest that polysaccharide coated particles are more individually distributed while agglomeration of the uncoated particles limits the surface area availability and access to membrane bound organelles.