Citrate-coated AgNPs Research Articles

An ultrasensitive "mix-and-detect" kind of fluorescent biosensor for malaoxon detection using the AChE-ATCh-Ag-GO system.

Malaoxon, a highly toxic metabolite of malathion, can lead to severe harm or death if ingested. This study introduces a rapid and innovative fluorescent biosensor that relies on acetylcholinesterase (AChE) inhibition for detecting malaoxon using Ag-GO nanohybrid. The synthesized nanomaterials (GO, Ag-GO) were evaluated with multiple characterization methods to confirm their elemental composition, morphology, and crystalline structure. The fabricated biosensor works by utilizing AChE to catalyze the substrate acetylthiocholine (ATCh), which generates positively charged thiocholine (TCh) and triggers citrate-coated AgNP aggregation on the GO sheet, leading to an increase in fluorescence emission at 423 nm. However, the presence of malaoxon inhibits the AChE action and reduces the production of TCh, resulting in a decrease in fluorescence emission intensity. This mechanism allows the biosensor to detect a wide range of malaoxon concentrations with excellent linearity and low LOD and LOQ values of 0.001 pM to 1000 pM, 0.9 fM, and 3 fM, respectively. The biosensor also demonstrated superior inhibitory efficacy towards malaoxon compared to other OP pesticides, indicating its resistance to external influences. In practical sample testing, the biosensor displayed recoveries of over 98% with extremely low RSD% values. Based on the results obtained from the study, it can be concluded that the developed biosensor has the potential to be used in various real-world applications for detecting malaoxon in food, and water samples, with high sensitivity, accuracy, and reliability.

Silver nanoparticles exert toxic effects in human monocytes and macrophages associated with the disruption of Δψm and release of pro-inflammatory cytokines.

Silver nanoparticles (AgNP) are the most widely produced type of nanoparticles due to their antimicrobial and preservative properties. However, their systemic bioavailability may be considered a potential hazard. When AgNP reach the bloodstream, they interact with the immune cells, contributing to the onset and development of an inflammatory response. Monocytes and macrophages play a pivotal role in our defense system, but the interaction of AgNP with these cells is still not clear. Therefore, the main objective of this work was to assess the cytotoxic and pro-inflammatory effects induced by 5, 10, and 50nm AgNP coated with polyvinylpyrrolidone (PVP) and citrate, in concentrations that could be attained in vivo (0-25μg/mL), in human monocytes isolated from human blood and human macrophages derived from a monocytic cell line (THP-1). The effects of PVP and citrate-coated AgNP on cell viability, mitochondrial membrane potential, and cytokines release were evaluated. The results evidenced that AgNP exert strong harmful effects in both monocytes and macrophages, through the establishment of a strong pro-inflammatory response that culminates in cell death. The observed effects were dependent on the AgNP concentration, size and coating, being observed more pronounced cytotoxic effects with smaller PVP coated AgNP. The results showed that human monocytes seem to be more sensitive to AgNP exposure than human macrophages. Considering the increased daily use of AgNP, it is imperative to further explore the adverse outcomes and mechanistic pathways leading to AgNP-induced pro-inflammatory effects to deep insight into the molecular mechanism involved in this effect.

Multispectroscopic studies on the molecular interactions between bovine γ-globulin and borohydride-capped silver nanoparticles.

Interactions between bovine γ-globulin (BGG) and borohydride-capped silver nanoparticles (BAgNPs) were studied using dynamic light scattering (DLS) and spectroscopic techniques such as UV-vis spectroscopy, fluorescence, and circular dichroism. The results were compared with earlier reported interactions between γ-globulin and citrate-coated AgNPs (CAgNPs). BAgNPs were synthesized and characterized. Irrespective of the coating on AgNPs, nanoparticles had formed ground-state complexes with the protein. CAgNPs, as well as BAgNPs had caused static quenching of tryptophan (Trp) fluorescence of the protein. The change in the capping agent from citrate to borohydride weakened the binding of nanoparticles with the protein. But the same change in capping agent had increased the fluorescence quenching efficiency of AgNPs. Hydrogen bonding and van der Waals interactions were involved in BGG-BAgNPs complex similar to the CAgNPs complex with γ-globulin. Polarity of the Trp microenvironment in BGG was not altered using BAgNPs as opposed to CAgNPs, as supported using synchronous and three-dimensional fluorescence. Resonance light scattering experiments also suggested nano-bio conjugation. Far-UV and near-UV circular dichroism (CD) spectra respectively pointed towards changes in the secondary and tertiary structure of BGG by BAgNPs, which was not observed for CAgNPs.

Silver Nanoparticles Induce Apoptosis in HepG2 Cells through Particle-Specific Effects on Mitochondria.

Silver nanoparticles (AgNPs) have been widely used in biomedical and consumer products. It remains challenging to distinguish the toxicity of AgNPs derived from the particle form or the released silver ions (Ag+). In this study, the toxic effects of two citrate-coated AgNPs (20 and 100 nm) and Ag+ were investigated in hepatoblastoma cells (HepG2 cells). The suppression tests showed that AgNPs and Ag+ induced cell apoptosis via different pathways, which led us to speculate on the AgNP-induced mitochondrial damage. Then, the mitochondrial damages induced by AgNPs and Ag+ were compared under the same intracellular Ag+ concentration, showing that the mitochondrial damage might be mainly attributed to Ag nanoparticles but not to Ag+. The interaction between AgNPs and mitochondria was analyzed using a scattered light imaging method combined with light intensity profiles and transmission electron microscopy. The colocalization of AgNPs and mitochondria was observed in both NP20- and NP100-treated HepG2 cells, indicating a potential direct interaction between AgNPs and mitochondria. These results together showed that AgNPs induced apoptosis in HepG2 cells through the particle-specific effects on mitochondria.

Evaluation of silver nanoparticle acute and chronic effects on freshwater amphipod (Hyalella azteca).

Silver nanoparticles (AgNPs) are known to cause ecotoxic effects, but there are no existing derived ambient water quality criteria (AWQC) for these nanomaterials to protect freshwater aquatic life due to insufficient toxicological data. We exposed Hyalella azteca to silver nitrate, citrate-coated AgNPs (citrate-AgNPs), and polyvinylpyrrolidone-coated AgNPs (PVP-AgNPs) in a 10-day and 28-day water-only static renewal system with clean sand as a substrate for the amphipods and compared their point estimates with the United States Environmental Protection Agency (USEPA) AWQC for silver. We observed that all treatments decreased the survival, growth, and biomass of H. azteca, and the order of toxicity was AgNO3 > citrate-AgNPs > PVP-AgNPs. The LC50s of AgNO3, citrate-AgNPs, and PVP-AgNPs were 3.0, 9.6, and 296.0µg total Ag L-1, respectively, for the acute exposure and 2.4, 3.2, and 61.4µg total Ag L-1, respectively, for the chronic exposure. Acute and chronic EC20s of citrate-AgNPs ranged from 0.5 to 3.5µg total Ag L-1 while that of PVP-AgNPs ranged from 31.2 to 175µg total Ag L-1 for growth and biomass. Both Ag+ released from AgNPs and the nanoparticles contributed to the observed toxicity. The dissolution and toxicity of AgNPs were influenced by surface coating agents, particle size, and surface charge. Most point estimates for AgNPs were above AWQC for silver (4.1µg L-1) and the lowest concentration (0.12µg/L) at which Ag is expected to cause chronic adverse effects to freshwater aquatic life. Our study demonstrates that the current AWQC for silver, in general, is protective of freshwater aquatic life against AgNPs tested in the present study.

Stability and Microbial Toxicity of Silver Nanoparticles under Denitrifying Conditions

While the antibacterial effect of silver nanoparticles (AgNPs) on environmentally beneficial microbes has drawn considerable attention, the stability and microbial toxicity of AgNPs in a system where nitrate reduction is the dominant terminal electron-accepting process remain understudied. Here, we explore the impact of citrate-coated AgNPs (cit-AgNPs) on the growth and metabolism of two metal-sensitive and one nonsensitive bacterial strains under denitrifying conditions. Dose-response analysis revealed that in contrast to the bacteriostatic effect exhibited at 1 ppm, 5 ppm cit-AgNPs were bactericidal to the metal-sensitive strains. It was observed that the growth of the cells initiated Ag(I) formation, and the supplement of chloride (2.7 mM) to the cultures substantially mitigated the bactericidal capacity of cit-AgNPs, indicating that AgNP dissolution to ionic Ag(I) played a key role in AgNP toxicity. Abiotic experiments confirmed that nitrite, not nitrate, had the capacity to oxidize cit-AgNPs. Transcriptomic analysis revealed that (i) the gene encoding for membrane stress was upregulated proportionally to cit-AgNP concentrations; (ii) cit-AgNPs and Ag(I) at higher levels upregulated genes involved in oxidative stress and iron-sulfur clusters, whereas expressions of the genes responsible for electron transport, ATP synthesis, and denitrification were substantially repressed; (iii) the addition of chloride significantly altered the level of transcriptional profiles of all of the genes. These results not only provide evidence of abiotic AgNP oxidation by metabolic intermediate nitrogen species but also suggest that AgNPs and Ag(I) may induce differential toxicity modes to prokaryotes. Our findings reinforce the importance of evaluating the potential ecological toxicity and risks associated with the transformation of nanomaterials.

Global gene expression signatures in response to citrate-coated silver nanoparticles exposure

Silver nanoparticles (AgNPs) are widely used in medical and commercial products for their unique antibacterial functions. However, the impact of AgNPs on human neural development is not well understood. To investigate the effect of AgNPs on human neural development, various doses of 20 nm citrate-coated AgNP (AgSC) were administered to human embryonic stem cell derived neural progenitors during the neuronal differentiation. Immunofluorescence staining with neuronal progenitor markers SOX2 (sex determining region Y-box 2) and Nestin (VI intermediate filament protein) showed that AgSC inhibited rosette formation, neuronal progenitor proliferation, and neurite outgrowth. Furthermore, AgSC promoted astrocyte activation and neuronal apoptosis. These adverse effects can be partially recovered with ascorbic acid. A genome-wide transcriptome analysis of both AgSC treated and untreated samples indicated that the most up-graduated genes were a group of Metallothionein (1F, 1E, 2A) proteins, a metal-binding protein that plays an essential role in metal homeostasis, heavy metal detoxification, and cellular anti-oxidative defence. The most significantly down-regulated genes were neuronal differentiation 6 (NEUROD6) and fork head box G1 (FOXG1). GO analyse indicated that the regulation of cholesterol biosynthetic process, neuron differentiation, synapse organization and pattern specification, oliogenesis, and neuronal apoptosis were the most impacted biological processes. KEGG pathway analyse showed that the most significantly impacted pathways were C5 isoprenoid, axon guidance, Notch, WNT, RAS-MAPK signalling pathways, lysosome, and apoptosis. Our data suggests that AgSCs interfered with metal homeostasis and cholesterol biosynthesis which induced oxidative stress, inhibited neurogenesis, axon guidance, and promoted apoptosis. Supplementation with ascorbic acid could act as an antioxidant to prevent AgSC-mediated neurotoxicity.

Disruption of brain conductivity and permittivity and neurotransmitters induced by citrate-coated silver nanoparticles in male rats.

As one of the most exonerative, competitive, and abundant nanoparticles in curative uses, silver nanoparticles (AgNPs) play a growing important role in developing global neurodegeneration. Herein, we inspected the neurotoxic and histopathological effects of the oral dose of 26.9 nm citrate-coated AgNPs (100 and 1000 mg/kgbw, 28 days) on the brain conductivity and permittivity combined with neurotransmitter assays. While male mice in the control group were given deionized water. In terms of biophysical levels, the brain electric conductivity and relative permittivity were significantly decreased in the 26.9 nm citrate-coated AgNP treated groups versus the controls. Besides, 26.9 nm citrate-coated AgNP treatment resulted in a significant deficiency in the concentrations of brain acetylcholine esterase, dopamine, and serotonin. Total brain contents of silver ion significantly increased in a dose-dependent manner. Further, light and electron microscopy revealed a progressive disruption in the lamellar pattern of the myelinated axons of the nerve fibers, in addition to the accumulation of nanosilver in lysosomes and swollen mitochondria in axoplasm. In conclusion, 26.9 nm citrate-coated AgNPs are capable of gaining access to the brain of mice and causing electric conductivity and relative permittivity damage along with a high degree of cellular toxicity in the brain tissue. Therefore, the present study highlights, for the first time, the adverse effects of the citrate-coated AgNPs to the brain of mice and raises the concern of their probable neurotoxic impacts which is helpful for conclusive interpretation of future behavioral and potential neurodegeneration-based aspects. It would be of interest to investigate citrate-coated AgNPs mediated axonal relevant-signal transduction levels in future studies.

Transformation and Cytotoxicity of Surface-Modified Silver Nanoparticles Undergoing Long-Term Aging.

Silver nanoparticles (AgNPs) are constituents of many consumer products, but the future of their production depends on ensuring safety. The stability of AgNPs in various physiological solutions and aging in storage may affect the accuracy of predicted nanoparticle toxicity. The goal of this study was to simulate the transformation of AgNPs in different media representatives to the life cycle in the environment and to identify their toxicity to Hepa1c1c7 cells in a long-term aging process. AgNPs coated with citrate, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), and branched polyethyleneimine (BPEI) were studied. Our results show that the exposure media had a significant impact on the transformation of AgNPs. Citrate-coated AgNPs showed significant aggregation in phosphate-buffered saline. The aging of AgNPs in optimal storage showed that the charge-stabilized particles (citrate) were more unstable, with significant aggregation and shape changes, than sterically stabilized particles (PEG AgNPs, PVP AgNPs). The BPEI AgNPs showed the highest dissolution of AgNPs, which induced significantly increased toxicity to Hepa1c1c7 cells. Overall, our findings showed that storage and media of AgNPs influenced the transformation of AgNPs and that the resulting changes in the AgNPs’ physicochemical properties influenced their toxicity. Our study contributes to the understanding of AgNPs’ transformations under realistic exposure scenarios and increasing the predictability of risk assessments.

Transcriptomics reveals the action mechanisms and cellular targets of citrate-coated silver nanoparticles in a ubiquitous aquatic fungus.

Silver nanoparticles (AgNPs) are among the major groups of contaminants of emerging concern for aquatic ecosystems. The massive application of AgNPs relies on the antimicrobial properties of Ag, raising concerns about their potential risk to ecologically important freshwater microbes and the processes they drive. Moreover, it is still uncertain whether the effects of AgNPs are driven by the same mechanisms underlying those of Ag ions (Ag+). We employed transcriptomics to better understand AgNP toxicity and disentangle the role of Ag+ in the overall toxicity towards aquatic fungi. To that end, the worldwide-distributed aquatic fungus Articulospora tetracladia, that plays a central role in organic matter turnover in freshwaters, was selected and exposed for 3 days to citrate-coated AgNPs (∼20nm) and Ag+ at concentrations inhibiting 20% of growth (EC20). Responses revealed 258 up- and 162 down-regulated genes upon exposure to AgNPs and 448 up- and 84 down-regulated genes under exposure to Ag+. Different gene expression patterns were found after exposure to each silver form, suggesting distinct mechanisms of action. Gene ontology (GO) analyses showed that the major cellular targets likely affected by both silver forms were the biological membranes. GO-based biological processes indicated that AgNPs up-regulated the genes involved in transport, nucleobase metabolism and energy production, but down-regulated those associated with redox and carbohydrate metabolism. Ag+ up-regulated the genes involved in carbohydrate and steroid metabolism, whereas genes involved in localization and transport were down-regulated. Our results showed, for the first time, distinct profiles of gene expression in aquatic fungi exposed to AgNPs and Ag+, supporting different modes of toxicity of each silver form. Also, our results suggest that Ag+ had a negligible role in the toxicity induced by AgNPs. Finally, our study highlights the power of transcriptomics in portraying the stress induced by different silver forms in organisms.

Proteomics reveals surface electrical property-dependent toxic mechanisms of silver nanoparticles in Chlorella vulgaris

Silver nanoparticles (AgNPs) are known to exert adverse effects on both humans and aquatic organisms; however, the toxic mechanisms underlying these effects remain unclear. In this study, we investigated the toxic mechanisms of various AgNPs with different surface electrical properties in the freshwater algae Chlorella vulgaris using an advanced proteomics approach with Data-Independent Acquisition. Citrate-coated AgNPs (Cit-AgNPs) and polyethyleneimine-coated AgNPs (PEI-AgNPs) were selected as representatives of negatively and positively charged nanoparticles, respectively. Our results demonstrated that the AgNPs exhibited surface electrical property-dependent effects on the proteomic profile of C. vulgaris. In particular, the negatively charged Cit-AgNPs specifically regulated mitochondrial function-related proteins, resulting in the disruption of several associated metabolic pathways, such as those related to energy metabolism, oxidative phosphorylation, and amino acid synthesis. In contrast, the positively charged PEI-AgNPs primarily targeted ribosome function-related proteins and interrupted pathways of protein synthesis and DNA genetic information transmission. In addition, Ag+ ions released from the AgNPs had a significant influence on protein regulation and the induction of cellular stress. Collectively, our findings provide new insight into the surface electrical property-dependent proteomic effects of AgNPs on C. vulgaris and should improve our understanding of the toxic mechanisms of AgNPs in freshwater algae.

Surface charge-dependent bioaccumulation dynamics of silver nanoparticles in freshwater algae

Silver nanoparticles (AgNPs) can gradually accumulate in algae to exert their toxicity; however, there is little knowledge about their bioaccumulation dynamics. For the first time, this study reports the effect of surface charge of AgNPs on their bioaccumulation dynamics in freshwater algae (Chlorella vulgaris) using biodynamic modeling. Polyethylene-coated AgNPs (PEI-AgNPs) and citrate-coated AgNPs (Cit-AgNPs) were selected as positively and negatively charged AgNPs, i.e., P-AgNPs and N-AgNPs, respectively. Their uptake and elimination dynamics were investigated at a concentration of 50% inhibition of growth rate values (EC50) and 10% inhibition of growth rate values (EC10). The one-component model can generally well simulate the algal uptake and elimination kinetics of N-AgNPs but not of P-AgNPs. At both concentrations, the uptake rate constants (ku) for P-AgNPs were ∼20 times higher than that for N-AgNPs. The parameters of biphasic elimination kinetics revealed that P-AgNPs were eliminated faster than N-AgNPs during depuration compared to in subsequent processes. Compared with N-AgNPs, extended Derjaguin–Landau–Verwey–Overbeek (DLVO) theory and dark-field imaging revealed that P-AgNPs can be rapidly absorbed on the algal cell surface membrane owing to their remarkably lower energy barrier between algal cells, resulting in a faster adsorption/uptake process and aggregation of algal cells. Our results clearly demonstrate that the AgNPs exhibited surface charge-dependent bioaccumulation dynamics in algal cells. Thus, AgNP surface charge primarily influences the AgNP accumulation dynamics in algal cells.

Fast dissolution of silver nanoparticles at physiological pH

While silver nanoparticles (AgNP) are used in topical treatments and medical devices for humans, no smooth, safe remedy exists to remove them and avoid possible post-treatment uptake in the body. We show here that cysteamine hydrochloride (CYS∙HCl), a simple FDA and EMA approved molecule, is able to dramatically accelerate the otherwise extremely slow oxidation of citrate-coated AgNP by O2 in a wide range of pH, including the physiological 7.4 value, obtaining the halving of AgNP concentration in t < 10 min. The dependence of oxidation kinetics on CYS concentration and pH is studied, finding faster processes on increasing CYS and basicity, despite the decrease of O2 reduction potential. Complexation and electrochemical studies demonstrate that CYS adhesion to AgNP surface followed by formation of 1:2 Ag+:CYS complex is the driving force for the AgNP oxidation, this also giving a definitive explanation to the otherwise still unclear phenomenon of AgNP etching by thiols. The efficacy of CYS∙HCl is verified also on AgNP coated with pectin and PEG-SH, and on AgNP immobilized on surfaces.

Environmental fate and behavior of silver nanoparticles in natural estuarine systems

Silver nanoparticles (AgNPs) are widely used in many consumer products, whereas their environmental behaviors in natural aquatic systems remain unknown, especially in natural brackish media. Therefore, it is urgent to investigate the environmental fate of AgNPs in natural brackish waters. Here, we investigated the stability of citrate-coated AgNPs in natural brackish water collected from 6 different sites with distinct salinities in the Xinglinwan Reservoir, located in Xiamen City, southeast China. The obtained results showed that AgNP colloids remained stable in low-salinity waters, which was mainly determined by the effects of dissolved organic matter (DOM) promoting the stability of the nanoparticles. However, the environmental fate of AgNPs in high-salinity waters was dominated by the salinity or ionic strength, especially the free ion concentrations of Cl−, SO42−, or S2−, resulting in rapid sedimentation and dissolution. In addition, both DOM and salinity contributed to the environmental behavior of AgNPs in moderate-salinity waters, ultimately resulting in either colloidal stability or sedimentation. Overall, these results may reveal that AgNPs remain relatively stable for a long period in low-salinity natural waters, and that the stability might gradually decrease as AgNPs are transferred from freshwaters through brackish waters and eventually end up in seawater along the bay. Our findings also further indicate that the toxicity and potential risks of AgNPs may present more serious threats to the environment and organisms in natural freshwaters than in natural estuarine systems or seawater.

Dissolution of Silver Nanoparticles in Colloidal Consumer Products: Effects of Particle Size and Capping Agent.

The utilization of silver nanoparticles (AgNPs) in consumer products has significantly increased in recent years, primarily due to their antimicrobial properties. Increased use of AgNPs has raised ecological concerns. Once released into an aquatic environment, AgNPs may undergo oxidative dissolution leading to the generation of toxic Ag+. Therefore, it is critical to investigate the ecotoxicological potential of AgNPs and determine the physicochemical parameters that control their dissolution in aquatic environments. We have investigated the dissolution trends of aqueous colloidal AgNPs in five products, marketed as dietary supplements and surface sanitizers. The dissolution trends of AgNPs in studied products were compared to the dissolution trends of AgNPs in well-characterized laboratory-synthesized nanomaterials: citrate-coated AgNPs, polyvinylpyrrolidone-coated AgNPs, and branched polyethyleneimine-coated AgNPs. The characterization of the studied AgNPs included: particle size, anion content, metal content, silver speciation, and capping agent identification. There were small differences in the dissolved masses of Ag+ between products, but we did not observe any significant differences in the dissolution trends obtained for deionized water and tap water. The decrease of the dissolved mass of Ag+ in tap water could be due to the reaction between Ag+ and Cl-, forming AgCl and affecting their dissolution. We observed a rapid initial Ag+ release and particle size decrease for all AgNP suspensions due to the desorption of Ag+ from the nanoparticles surfaces. The observed differences in dissolution trends between AgNPs in products and laboratory-synthesized AgNPs could be caused by variances in capping agent, particle size, and total AgNP surface area in suspensions.

Interaction of silver nanoparticles with mediterranean agricultural soils: Lab-controlled adsorption and desorption studies

The production of silver nanoparticles (AgNPs) has increased tremendously during recent years due to their antibacterial and physicochemical properties. As a consequence, these particles are released inevitably into the environment, with soil being the main sink of disposal. Soil interactions have an effect on AgNP mobility, transport and bioavailability. To understand AgNP adsorption processes, lab-controlled kinetic studies were performed. Batch tests performed with five different Mediterranean agricultural soils showed that cation exchange capacity and electrical conductivity are the main parameters controlling the adsorption processes. The adsorption kinetics of different sized (40, 75, 100 and 200 nm) and coated (citrate, polyvinylpyrrolidone and polyethyleneglycol (PEG)) AgNPs indicated that these nanoparticle properties have also an effect on the adsorption processes. To assess the mobility and bioavailability of AgNPs and to determine if their form is maintained during adsorption/desorption processes, loaded soils were submitted to leaching tests three weeks after batch adsorption studies. The DIN 38414-S4 extraction method indicated that AgNPs were strongly retained on soils, and single-particle inductively coupled plasma mass spectrometry confirmed that silver particles maintained their nanoform, except for 100 nm PEG-AgNPs and 40 nm citrate-coated AgNPs. The DTPA (diethylenetriaminepentaacetic acid) leaching test was more effective in extracting silver, but there was no presence of AgNPs in almost all of these leachates.

Determination of silver nanoparticle dose in vitro

An important issue for interpreting in vitro nanomaterial testing is quantifying the dose delivered to target cells. Considerations include the concentration added to the culture, the proportion of the applied dose that interacts with the target cells, and the amount that is eventually absorbed by the target cells. Rapid and efficient techniques are needed to determine delivered doses. Previously, we demonstrated that TiO2 and silver nanoparticles (AgNP) were absorbed by cells in a dose dependent manner between 1 μg/ml and 30 μg/ml and were detected in cells by light scatter using a flow cytometer. Here, we compare four potential indices of the dose of AgNP to cells, including: inductively coupled plasma - mass spectrometry (ICP-MS); flow cytometry side scatter (SSC); and amount of silver deposited to the cell layer as estimated with both an integrated Volumetric Centrifugation Method - In Vitro Sedimentation, Diffusion and Dosimetry Model (VCM-ISDD) and a Distorted Grid (DG) model. A retinal pigment epithelial cell line was exposed to 20 nm or 75 nm citrate-coated AgNP for 24 h. The relationships between particle sizes and internalized doses varied according to the dose metric. Twenty-four hours after exposure, the cell layer contained a greater mass of silver when treated with 75 nm AgNP than with 20 nm AgNP. When the dose was expressed as the number of particles or as the total surface area of absorbed particles, however, the reverse was true; the dose to the cells was higher after exposure to 20 than 75 nm AgNP. Flow cytometry SSC increased with dose for both sizes of AgNP, and was correlated with Ag in cells measured by ICP-MS. The rate of SSC increase was greater for 75 than for 20 nm AgNP, suggesting it could be used as an indicator of cellular dose after accounting for particle size and composition. Silver was detected by ICP-MS in re-suspended supernates of the isolated cell layer suggested that not all the silver deposited to the cell layer was absorbed by the cells. Both the VCM-ISDD and DG models estimated the proportion of Ag deposited to the cellular layer, which in both cases was greater than the amount of silver in the cells measured by ICP-MS. Modeled deposition more closely compared to the total Ag deposition by ICP-MS, i.e. mass of silver in the cells plus the resuspended, unabsorbed Ag from the cell layer. ICP-MS indicated the mass of silver in cells from AgNP treatment, but not whether the Ag was in the form of particles or dissolved ions. Deposition models predicted the amount of AgNP deposited to the cell layer, but not cellular uptake. Flow cytometry SSC was correlated to cellular uptake of particle-form AgNP and could be calibrated against ICP-MS to indicate mass of cellular uptake. Therefore, a combination of approaches may be required to accurately understand cellular dosimetry of in vitro nanotoxicology experiments. In summary, cellular dosimetry is an important consideration for nanotoxicology experiments, and not necessarily related to the applied dose.

Does plant growing condition affects biodistribution and biological effects of silver nanoparticles?

Among the many different types, silver nanoparticles (AgNPs) are the most commercialized and applied engineered nanoparticles in a wide range of areas, including agriculture. Despite numerous studies on their safety and toxicity of AgNPs, data on their effect and interactions with terrestrial plants are largely unknown. This study aimed to investigate the effect of growing conditions on the response of pepper plants (Capsicum annuum L.) to citrate-coated AgNPs. Growth parameters, biodistribution, and defence response were examined in peppers grown hydroponically or in soil substrate. In addition, the effects of nano and ionic form of silver were compared. The leaves and stems of peppers grown in substrate showed a higher bioaccumulation compared to hydroponically cultivated plants. The nano form of silver accumulated to a higher extent than ionic form in both leaves and stems. Both silver forms inhibited pepper growth to a very similar extent either through hydroponic or substrate growing settings. Unlike other studies, which investigated the effects of unrealistically high doses of AgNPs on different plant species, this study revealed that vascular plants are also susceptible to very low doses of AgNPs. Both silver forms affected all parameters used to evaluate oxidative stress response in pepper leaves; plant pigment and total phenolics contents were decreased, while lipid peroxidation and hydrogen peroxide lever were increased in treated plants. Similar biological effects of both nano and ionic Ag forms were observed for both substrate and hydroponic growing systems.

Silver nanoparticle immunomodulatory potential in absence of direct cytotoxicity in RAW 264.7 macrophages and MPRO 2.1 neutrophils

Engineered nanomaterials (ENM) are being used in a wide range of consumer products and pharmaceuticals; hence, there is an increasing risk for human exposure and potential adverse outcomes. The immune system, vital in host defense and protection against environmental agents, is typically initiated and executed by innate effector immune cells including macrophages and neutrophils. Previous literature has reported the immune system as a major target of ENM toxicity; however, there is inconsistency regarding the immunotoxicity of ENM. This could be attributed to differences in ENM physicochemical properties, cellular models examined, biocorona formation, etc. Thus, the current study examined the toxicity and immunomodulatory effects of silver nanoparticles (AgNP), one of the most utilized ENM in consumer and medical products, in two key innate immune cell models, e.g. RAW 264.7 cells (macrophages) and differentiated MPRO 2.1 cells (promyelocytes/neutrophils). The results showed that despite a generation of reactive oxygen species, exposure to 20 nm citrate-coated AgNP was not associated with major oxidative damage, inflammatory responses, nor cytotoxicity. Nevertheless, and most importantly, pre-exposure to the AgNP for 24 h enhanced RAW 264.7 cell phagocytic ability as well as the release of inflammatory cytokine interleukin-6 in response to lipopolysaccharide (LPS). In MPRO 2.1 cells, AgNP pre-exposure also resulted in enhanced phagocytic ability; however, these cells manifest reduced cell degranulation (elastase release) and oxidative burst in response to phorbol myristate acetate (PMA). Taken together, these findings indicated to us that exposure to AgNP, despite not being directly (cyto)toxic to these cells, had the potential to alter immune cell responses. The findings underscore the import of assessing immune cell function post-exposure to ENM beyond the standard endpoints such as oxidative stress and cytotoxicity. In addition, these findings further illustrate the importance of understanding the underlying molecular mechanisms of ENM-cellular interactions, particularly in the immune system.

Toxicity responses of different organs of zebrafish (Danio rerio) to silver nanoparticles with different particle sizes and surface coatings

Silver nanoparticles (AgNPs) in aquatic ecosystems are toxic to aquatic organisms. In this study, we aimed to investigate the toxicities and molecular mechanisms of AgNPs with different surface coatings (sodium citrate and polyvinylpyrrolidone) and particle sizes (20 nm and 100 nm) in the gills, intestines, and muscles of zebrafish after 96 h of exposure. Our results indicated that the contribution of particle size to AgNP toxicity was greater than that of the surface coating. Citrate-coated AgNPs were more toxic than polyvinylpyrrolidone-coated AgNPs, and 20-nm AgNPs were more toxic than 100-nm AgNPs. The toxic effects of AgNPs to the tissues were in the order intestines > gills > muscles. Differential expression of genes with the different AgNPs confirmed that they had toxic effects in the zebrafish tissues at the molecular level. Our comprehensive comparison of the toxicities of different AgNPs to aquatic ecosystems will be helpful for further risk assessments of AgNPs.