Dissolved Silver Ions Research Articles

Exploring the impact of silver-based nanomaterial feed additives on green algae through single-cell techniques

Silver in its various forms, including dissolved silver ions (Ag+) and silver nanoparticles (AgNPs), is a promising alternative to traditional antibiotics, largely used in livestock as feed additives and could contribute to the decrease and avoidance of the development of antibiotic resistance. The present study aims to assess the potential ecotoxicity of a silver-based nanomaterial (Ag-kaolin), the feed supplemented with the nanomaterial and the faeces since the latter are the ones that finally reach the environment. To this end, green alga Raphidocellis subcapitata was exposed to the extracts of Ag-kaolin, supplemented feed, and pig faeces for 72 h, along with Ag+ and AgNPs as controls for comparison purposes. Given the complexity of the studied materials, single-cell techniques were used to follow the changes in the cell numbers and chlorophyll fluorescence by flow cytometry, and the accumulation of silver in the exposed cells by single cell inductively coupled plasma mass spectrometry (SC-ICP-MS). Changes in cell morphology were observed by cell imaging multimode reader. The results revealed a decrease in chlorophyll fluorescence, even at low concentrations of Ag-kaolin (10 μg L−1) after 48 h of exposure. Additionally, complete growth inhibition was found with this material like the results obtained by exposure to Ag+. For the supplemented feed, a concentration of 50 μg L−1 was necessary to achieve complete growth inhibition. However, the behaviour differed for the leachate of faeces, which released Ag2S and AgCl alongside Ag+ and AgNPs. At 50 μg L−1, inhibition was minimal, primarily due to the predominance of less toxic Ag2S in the leachate. The uptake of silver by the cells was confirmed with all the samples through SC-ICP-MS analysis. These findings demonstrate that the use of Ag-kaolin as a feed supplement will lead to a low environmental impact.

Antibacterial Activity of Dissolved Silver Fractions Released from Silver-Coated Titanium Dental Implant Abutments: A Study on Streptococcus mutans Biofilm Formation.

(1) Background: The aim of this research was to investigate the antibacterial activity of dissolved silver from silver-coated titanium implants against Streptococcus mutans. (2) Methodology: Silver-coated titanium implant discs were immersed in 1.8 mL of brain heart infusion broth (BHIB) and incubated for 24 h in order to release the silver ions into the broth. The coating quality was confirmed via EDS, and the dissolved silver was measured via inductively coupled plasma mass spectrometry (ICP-MS). The experimental design used unconditioned broth (control) and broth conditioned with silver released from silver-coated titanium implants (n = 6). Regarding the antibacterial activity, isolated Streptococcus mutans was used. A turbidity test and lactate production test were performed to determine the effect of dissolved silver on bacterial growth in a suspension and biofilm formation. (3) Result: The results showed that the coating was successfully applied on the substrate. There was around 0.3 mg/L of silver released into the BHIB, and the turbidity of the control group was significantly higher than the treatment, with measured absorbance values of 1.4 and 0.8, respectively, indicating that the dissolved silver ions from the silver-coated titanium discs exhibited some degree of antibacterial activity by preventing the growth of Streptococcus mutans. However, the results of the antibiofilm activity test did not show any significant difference between the groups. (4) Conclusion: The dissolved silver from silver-coated titanium implants has an antibacterial activity but not a significant antimicrobial activity, indicating that the dissolved silver from silver-coated titanium abutments can significantly reduce the incidence of peri-implant mucositis.

Characterizing silver nanoparticles in beverages and following their release from silver-containing food containers using sector field single particle inductively coupled plasma mass spectrometry (SP-ICP-MS)

Silver nanoparticles (Ag NPs) are increasingly used in the food industry because of their excellent antibacterial activity. They are integrated into coatings of various food packaging to help ensure longer product shelf life. Nonetheless, the risks associated with Ag NPs are currently not well known and their potential effects on humans are causing growing concern. Furthermore, it is not clear whether NPs have greater or lesser risk than dissolved silver ions or bulk phase Ag. Consequently, it is necessary to characterize the release of silver from silver-enhanced containers into food matrices using sensitive analytical techniques that allow one to distinguish between silver ions and nanoparticles. Single particle ICP-MS is a promising technique to quantify and determine size distributions of very small particles at low concentrations and distinguish between ionic and particulate forms of the analyte. This study focused first on the aging of Ag NPs in three different food simulants (Milli-Q water, 10% ethanol and 3% acetic acid) and in two beverages (orange juice and infant milk formula) in order to validate the technique for low NP concentrations. Acidic media caused significant NP oxidation while the organic macromolecules in the juice appeared to increase the stability of the NPs. Subsequently, a migration study from silver-enhanced containers showed that release was mainly due to ionic forms of the silver. Significant release of dissolved Ag occurred in 3% acetic acid (58% of the total Ag mass of the containers) with lower release being observed in Milli-Q water (33%), 10% ethanol (29%), orange juice (17%) and milk formula (5%). Heating led to a considerable release of silver (again dissolved), likely because of the swelling of the polymer containers. The study suggests that the risks to foods associated with Ag NP in packaging can be determined using existing regulations and guidelines based on (ionic) Ag.

Effect of organic matter on the trophic transfer of silver nanoparticles in an aquatic food chain

The behavior and toxicity of nanoparticles could be affected significantly by the ubiquitous natural organic matter (NOM) in aquatic environments. However, the influence of NOM on nanoparticles along the food chain remains largely unknown. This study constructed bacteria Escherichia coli (E. coli) - protozoa Tetrahymena thermophila (T. thermophila) to evaluate the influence of NOM on the bioaccumulation, trophic transfer and toxicity of silver nanoparticles (Ag NPs). Results demonstrated that NOM could reduce the toxicity of Ag NPs to E. coli and T. thermophila by different influence mechanisms (e.g., reduce Ag NPs accumulation or complex with dissolved silver ion (Ag+)) which related to the type of NOM and organisms. Moreover, Ag NPs can be transferred and biomagnified to T. thermophila via trophic transfer. Three typical NOM could significantly increase the trophic transfer factors of Ag NPs ranging from 1.16 to 2.49, which may be ascribed to NOM reducing the capacity for T. thermophila to excrete total silver (Ag) as NOM could significantly change the form of Ag. These findings provide a novel insight into the impact of NOM on the ecological risk posed by Ag NPs through the food chain and emphasize the need to understand further the interactions between nanoparticles and NOM in various ecosystems.

Transcriptomic responses to silver nanoparticles in the freshwater unicellular eukaryote Tetrahymena thermophila.

Currently, silver nanoparticles (AgNPs) are being increasingly used as biocides in various consumer products and if released in the environment they can affect non-target organisms. Therefore, understanding the toxicity mechanism is crucial for both the design of more efficient nano-antimicrobials and for the design of nanomaterials that are biologically and environmentally benign throughout their life-cycle. Here, the ciliate Tetrahymena thermophila was used to elucidate the mechanisms of action of AgNPs by analysing the gene expression profile by RNA-seq and the transcriptomic effects of AgNPs were compared to those induced by soluble silver salt, AgNO3. Exposure to AgNPs at sublethal concentrations for 24h induced phagocytosis, transport pathways, response to oxidative stress, glutathione peroxidase activity, response to stimulus, oxidation-reduction, proteolysis, and nitrogen metabolism process. Based on gene set enrichment analysis (GSEA), some biological processes appeared targets of both toxicants. In addition to many similarities in affected genes, some effects were triggered only by NPs, like phagocytosis, glutathione peroxidase activity, response to stimulus, protein phosphorylation and nitrogen metabolism process. This research provides evidence that AgNPs compared to AgNO3 at the same concentration of dissolved silver ions dysregulate a higher number of cellular pathways. These findings confirm that AgNPs can induce toxicity not only due to soluble silver ions released from the particles but also to particle intrinsic features.

Synthesis, dissolution, and regeneration of silver nanoparticles stabilized by tannic acid in aqueous solution

Silver nanoparticles (AgNPs), especially with small size, are easy to release silver ion in aqueous solution owing to various reasons, which would significantly affect the stability, properties, and application of AgNPs. In this paper, monodisperse AgNPs with small size of ca. 10 nm were successfully prepared based on solid-state reactions. Ascorbic acid (AA) was used as reductant and tannic acid (TA) was used both as reductant and stabilizer in this environmentally friendly reaction. The dissolution and regeneration of the as-prepared TA-AgNPs in pure water were investigated by UV−vis spectra, TEM observations, and differential pulse anodic stripping voltammetry. The results indicated that the TA-AgNPs showed a little higher dissolution than conventional PVP-coated ones with similar size. However, the dissolved silver ion in the TA-AgNPs aqueous solution could be recovered just by adjusting the pH of the solution, which could be attributed to the reductant performance of TA at alkaline conditions. After regeneration, some smaller nanoparticles appeared in TA-AgNPs aqueous solution, indicating that new nucleation formed and the dissolved silver ions were actually recovered to Ag0.

Role of extracellular polymeric substances on the behavior and toxicity of silver nanoparticles and ions to green algae Chlorella vulgaris

The effect of extracellular polymeric substances (EPS), vital organic matters and nutrient elements in the natural environment, on the behavior and toxicology of silver nanoparticles (AgNPs) and ions remains ambiguous. In this study, the role of EPS on the toxicity of AgNPs and dissolved silver ions (from AgNO3) to a green algae Chlorella vulgaris was investigated. After the removal of EPS, algae accumulated more silver, about 7.41- and 1.25-fold of those in the algae with EPS for AgNPs and AgNO3 treatments, respectively. The large amount of accumulated silver was bound to the algal cell surface for AgNPs treatment and was internalized in the algae for AgNO3 treatment, irrespective of the presence of EPS in algae. After exposure to AgNPs, the ruffles in the surfaces of algal cells were filled by AgNPs, and almost invisible. FTIR showed that for both AgNPs and AgNO3, the aldehyde groups on the cell surface were oxidized to carboxyl groups by silver ions, irrespective of the presence of EPS in algal cells, indicating that silver ions were released from the oxidization of AgNPs and reacted with algal cells. The content of chlorophyll showed that AgNPs depressed algal growth more remarkably than did AgNO3, independent of the presence of EPS in algae, suggesting that AgNPs had greater toxic effects on algae than did silver ions. The findings suggest that the barrier effect of EPS gave nanoparticles an extraordinary edge over ions, but EPS had no discerning effect on the interaction of algal cells with the silver ions released from AgNPs and AgNO3, and also on the effect of AgNPs and AgNO3 on algal growth.

Visualization of Silver Nanoparticle Formation on Nanoscale Zero-Valent Iron

Instant reactions of Ag(I) on nanoscale zero-valent iron (nZVI) were visualized with spherical-aberration-corrected scanning transmission electron microscopy (Cs-STEM) integrated with X-ray energy dispersive spectroscopy. In particular, the rapid growth of pure silver nanoparticles on nZVI was captured with high-resolution STEM imaging. Dissolved silver ions are attracted to the negatively charged iron nanoparticles and quickly reduced to metallic silver on the defective and electron-rich iron surface. Small (∼1–2 nm) and highly reactive silver nanoparticles are formed along the long chains of nZVI. Results reinforce the applications of nZVI for rapid enrichment, complete separation, and recovery of precious metals such as Au and Ag from trace-level sources such as wastewater and demonstrate the potential of analytical electron microscopy for mapping pollutant reactions on the surface of nanoparticles.

Humic acid attenuation of silver nanoparticle toxicity by ion complexation and the formation of a Ag3+ coating

The use of silver nanoparticles (AgNPs) result in an inevitable contact with aquatic environments. Here we study the behavior of AgNPs and the developmental toxicity in zebrafish embryos exposed to these nanoparticles (0–10 mg/L) with and without the presence of HA (20 mg/L), using zebrafish facility water (ZFW) and zebrafish growing media (ZGM). The presence of cations and HA gave rise to a decrease in Ag ion release and ζ-potential, an increase in the hydrodynamic diameter and oxidation of the AgNP surface. The results show that the presence of HA and cations in the media, as well as the silver speciation, i.e., the unusual presence of Ag3+, decreases the toxicity of AgNPs (LC50AgNPs: 1.19 mg/L; LC50AgNPs + HA: 3.56 mg/L), as well as silver bioavailability and toxicity in zebrafish embryos. Developmental alterations and the LC50 (1.19 mg/L) of AgNPs in ZFW were more relevant (p ≤ 0.05) than for AgNPs in ZGM (LC50 ˃ 10 mg/L). It was demonstrated that the bioaccumulation and toxicity of AgNPs depends on several factors including AgNPs concentration, nanoparticle aggregation, dissolved silver ions, speciation of silver ions, the amount of salt in the environment, the presence of humic substances and others, and different combinations of all of these factors.

Halide Welding for Silver Nanowire Network Electrode.

We developed a method of chemically welding silver nanowires (AgNWs) using an aqueous solution containing sodium halide salts (NaF, NaCl, NaBr, or NaI). The halide welding was performed simply by immersing the as-coated AgNW film into the sodium halide solution, and the resulting material was compared with those obtained using two typical thermal and plasmonic welding techniques. The halide welding dramatically reduced the sheet resistance of the AgNW electrode because of the strong fusion among nanowires at each junction while preserving the optical transmittance. The dramatic decrease in the sheet resistance was attributed to the autocatalytic addition of dissolved silver ions to the nanowire junction. Unlike thermal and plasmonic welding methods, the halide welding could be applied to AgNW films with a variety of deposition densities because the halide ions uniformly contacted the surface or junction regions. The optimized AgNW electrodes exhibited a sheet resistance of 9.3 Ω/sq at an optical transmittance of 92%. The halide welding significantly enhanced the mechanical flexibility of the electrode compared with the as-coated AgNWs. The halide-welded AgNWs were successfully used as source-drain electrodes in a transparent and flexible organic field-effect transistor (OFET). This simple, low-cost, and low-power consumption halide welding technique provides an innovative approach to preparing transparent electrodes for use in next-generation flexible optoelectronic devices.

New Insights into the Stability of Silver Sulfide Nanoparticles in Surface Water: Dissolution through Hypochlorite Oxidation.

Silver sulfide nanoparticles (Ag2SNPs) are considered to be stable in the environment due to the extreme low solubility of Ag2S (Ksp: 6.3 × 10-50). Little is known about the stability of Ag2SNPs in surface water disinfected with aqueous chlorine, one of the globally most used disinfectants. Our results suggested that both uncoated and polyvinylpyrrolidone (PVP)-coated Ag2SNPs (100 μg/L) underwent dissolution in surface water disinfected with aqueous chlorine at a dose of 4 mg/L, showing the highest dissolved silver ion concentrations of 22.3 and 10.5 μg/L within 45 min, respectively. The natural organic matter (NOM) and dissolved oxygen (DO) posed effects on the Ag2SNPs dissolution by chlorine; NOM accelerated Ag2SNPs dissolution while DO reduced the rate and extent of Ag2SNPs dissolution. We further demonstrated that Ag2SNPs dissolution was primarily attributed to active oxidative substances including hydroxyl radical and H2O2 originating from the hypochlorite oxidation. Additionally, water containing Ag2SNPs disinfected with hypochlorite showed stronger interference on the zebra fish (Danio rerio) embryo hatching than Ag2SNPs and hypochlorite on their own. This work documented that Ag2SNPs could undergo dissolution in surface water through hypochlorite oxidation, posing potential risks to aquatic organisms, and therefore showed new insights into the stability of Ag2SNPs in natural environment.

Application of thermospray flame furnace atomic absorption spectrometry for investigation of silver nanoparticles.

Thermospray flame furnace atomic absorption spectrometry (TS-FF-AAS) was applied to investigate the time-dependent absorption peak profile of various forms of silver. The thermospray flame furnace was set up with a 10-cm-long nickel tube with six holes, each 2.0mm in diameter, to allow the flame to enter, and this nickel tube acted as a furnace. A sample of 300μL was introduced into this furnace by use of water as a carrier at a flow rate of 0.5mLmin-1 through the ceramic capillary (0.5-mm inner diameter and 2.0-mm outer diameter), which was inserted into the front hole of the nickel tube. The system was applied to examine atomization behaviors of silver nanoparticles (AgNPs) with particle sizes ranging from 10 to 100nm. The atomization rate of AgNPs was faster than that of the dissolved silver ion. With increased amount of silver, the decay time observed from the time-dependent absorption peak profile was shortened in the case of dissolved silver ion, but it was increased in the case of AgNPs. With the particle size ranging from 10 to 100nm, the detection sensitivity was indirectly proportional to the particle size, suggesting that TS-FF-AAS may offer insights into the particle size of AgNPs provided that the concentration of the silver is known. To obtain quantitative information on AgNPs, acid dissolution of the particles was performed before TS-FF-AAS analysis, and recoveries of 80-110% were obtained.

Optical transformation of a CdTe quantum dot-based paper sensor for a visual fluorescence immunoassay induced by dissolved silver ions.

This work designs a simple low-cost visual fluorescence immunoassay for disease-related biomarkers (carcinoembryonic antigen, CEA, used as a model protein) in biological fluids, based on the structural and optical transformation of CdTe quantum dots (QDs) immobilized on paper induced by dissolved silver ions (Ag+) from silver nanoparticles (AgNPs) via a cation-exchange reaction. A sandwich-type immunoreaction was initially carried out in a removable polystyrene high-binding microplate coated with monoclonal anti-CEA capture antibody by using AgNP-labeled polyclonal anti-CEA antibody as the detection antibody. Thereafter, the carried AgNPs accompanying the sandwiched immunocomplexes were dissolved by acid to release numerous silver ions, which induced the ion-exchange reaction with the immobilized CdTe QDs on the paper (attached onto the microplate lid) for the Cd-to-Ag transformation, thus resulting in the quenching of the visual fluorescence from the CdTe QDs owing to Ag2Te formation. Under optimal conditions, the fluorescence intensity decreased with the increasing CEA concentration from 0.02 to 50 ng mL-1 with a detection limit of 5.6 pg mL-1. Further, a visual assay based on a CdTe QD-based paper sensor was developed for CEA detection, and 5.0 pg mL-1 CEA could be discriminated with the naked eye. In addition, our strategy displayed high specificity, good reproducibility and acceptable accuracy for analyzing human serum specimens with consistent results obtained using the commercialized enzyme-linked immunosorbent assay (ELISA) method.

Distinct transcriptomic responses of Caenorhabditis elegans to pristine and sulfidized silver nanoparticles

Manufactured nanoparticles (MNP) rapidly undergo aging processes once released from products. Silver sulfide (Ag2S) is the major transformation product formed during the wastewater treatment process for Ag-MNP. We examined toxicogenomic responses of pristine Ag-MNP, sulfidized Ag-MNP (sAg-MNP), and AgNO3 to a model soil organism, Caenorhabditis elegans. Transcriptomic profiling of nematodes which were exposed at the EC30 for reproduction for AgNO3, Ag-MNP, and sAg-MNP resulted in 571 differentially expressed genes. We independently verified expression of 4 genes (numr-1, rol-8, col-158, and grl-20) using qRT-PCR. Only 11% of differentially expressed genes were common among the three treatments. Gene ontology enrichment analysis also revealed that Ag-MNP and sAg-MNP had distinct toxicity mechanisms and did not share any of the biological processes. The processes most affected by Ag-MNP relate to metabolism, while those processes most affected by sAg-MNP relate to molting and the cuticle, and the most impacted processes for AgNO3 exposed nematodes was stress related. Additionally, as observed from qRT-PCR and mutant experiments, the responses to sAg-MNP were distinct from AgNO3 while some of the effects of pristine MNP were similar to AgNO3, suggesting that effects from Ag-MNP is partially due to dissolved silver ions.

Inhibition of bacterial surface colonization by immobilized silver nanoparticles depends critically on the planktonic bacterial concentration

Immobilization of antimicrobial silver nanoparticles (AgNPs) on surfaces has been proposed as a method to inhibit biofouling or as a possible route by which incidental releases of AgNPs may interfere with biofilms in the natural environment or in wastewater treatment. This study addresses the ability of planktonic Pseudomonas fluorescens bacteria to colonize surfaces with pre-adsorbed AgNPs. The ability of the AgNP-coated surfaces to inhibit colonization was controlled by the dissolved silver in the system, with a strong dependence on the initial planktonic cell concentration in the suspension, i.e., a strong inoculum effect. This dependence was attributed to a decrease in dissolved silver ion bioavailability and toxicity caused by its binding to cells and/or cell byproducts. Therefore, when the initial cell concentration was high (∼1×107CFU/mL), an excess of silver binding capacity removed most of the free silver and allowed both planktonic growth and surface colonization directly on the AgNP-coated surface. When the initial cell concentration was low (∼1×105CFU/mL), 100% killing of the planktonic cell inoculum occurred and prevented colonization. When an intermediate initial inoculum concentration (∼1×106CFU/mL) was sufficiently large to prevent 100% killing of planktonic cells, even with 99.97% initial killing, the planktonic population recovered and bacteria colonized the AgNP-coated surface. In some conditions, colonization of AgNP-coated surfaces was enhanced relative to silver-free controls, and the bacteria demonstrated a preferential attachment to AgNP-coated, rather than bare, surface regions. The degree to which the bacterial concentration dictates whether or not surface-immobilized AgNPs can inhibit colonization has significant implications both for the design of antimicrobial surfaces and for the potential environmental impacts of AgNPs.

Phytotoxicity, uptake, and accumulation of silver with different particle sizes and chemical forms

The antimicrobial property of silver nanoparticles (AgNPs) makes it one of the most commonly encountered nanomaterials in commercial products. Consequently, its detection in the environment is highly likely and its potential toxicity has been heavily investigated. While it is now generally agreed that AgNP itself exerts unique toxicity to plants in addition to that of dissolved silver ion, the accumulation and fate of different forms of silver in plant tissues are unknown. This study investigates the phytotoxicity, accumulation, and transport of Ag with different physical and chemical characteristics (e.g., ionic, nanoparticles, and bulk) in two agricultural crop species: Glycine max (soybean) and Triticum aestivum (wheat). The results showed that different forms of Ag demonstrated differential toxicity in these two species, with the Ag+ at the same nominal concentration displaying the strongest effect on plant growth. Exposure to 5 mg/L of elemental Ag in different forms all resulted in significant deposition on the root surface but its morphology and distribution patterns varied considerably. The Ag transport efficiency from roots to shoots differed with both Ag type and plant species. Notably, the upward transport of AgNPs (20–50 nm) was considerably more substantial than that of bulk Ag (1–3 µm). Cell fractionation studies confirmed that all types of Ag were internalized, with the plant cell wall as the predominant place for element accumulation. The findings demonstrate that Ag toxicity and in planta fate vary with particle type and that such considerations are likely necessary to adequately assess food safety concerns upon NP exposure.

Silver's salty twist on water splitting

Electrochemistry When plants split water during photosynthesis, they expel oxygen as a by-product. Ironically, it's this oxygen-generating step that's also proving the most difficult to optimize in artificial photosynthetic schemes, where the practical goal is simply to release the oxygen while accumulating hydrogen for fuel. Du et al. explore an alternative scheme, whereby the electrons to make the hydrogen come from chloride ions—already abundant in seawater—instead of other water molecules. Specifically, they show that dissolved silver ions act as efficient electrocatalysts for chloride oxidation, provided that the chloride is present at a high enough concentration to form (AgCl2)− and (AgCl3)2− ions, both of which are more soluble and more reactive than AgCl. J. Am. Chem. Soc. 10.1021/jacs.5b00037 (2015).

Ultrasensitive detection of single nucleotide polymorphism in human mitochondrial DNA utilizing ion-mediated cascade surface-enhanced Raman spectroscopy amplification.

Although surface-enhanced Raman spectroscopy (SERS) has been featured by high sensitivity, additional signal enhancement is still necessary for trace amount of biomolecules detection. In this paper, a SERS amplified approach, featuring "ions-mediated cascade amplification (IMCA)", was proposed by utilizing the dissolved silver ions (Ag(+)) from silver nanoparticles (AgNPs). We found that using Ag(+) as linkage agent can effectively control the gaps between neighboring 4-aminobenzenethiol (4-ABT) encoded gold nanoparticles (AuNPs@4-ABT) to form "hot spots" and thus produce SERS signal output, in which the SERS intensity was proportional to the concentration of Ag(+). Inspired by this finding, the IMCA was utilized for ultrasensitive detection of single nucleotide polymorphism in human mitochondrial DNA (16189T → C). Combining with the DNA ligase reaction, each target DNA binding event could successfully cause one AgNP introduction. By detecting the dissolved Ag(+) from AgNPs using IMCA, low to 3.0 × 10(-5) fm/μL targeted DNA can be detected, which corresponds to extractions from 200 nL cell suspension containing carcinoma pancreatic β-cell lines from diabetes patients. This IMCA approach is expected to be a universal strategy for ultrasensitive detection of analytes and supply valuable information for biomedical research and clinical early diagnosis.

The predominant species of ionic silver in biological media is colloidally dispersed nanoparticulate silver chloride

We have investigated the behaviour of silver ions in biologically relevant concentrations (10 to 100 ppm) in different media, from physiological salt solution over phosphate-buffered saline solution to protein-containing cell culture media. The results show that the initially present silver ions are bound as silver chloride due to the presence of chloride. Only in the absence of chloride, glucose is able to reduce Ag+ to Ag0. The precipitation of silver phosphate was not observed in any case. We conclude that the predominant silver species in biological media is dispersed nanoscopic silver chloride, surrounded by a protein corona which prevents the growth of the crystals and leads to colloidal stabilization. Therefore, in cell culture experiments where dissolved silver ions are studied in the upper ppm range, in fact the effect of colloidally dispersed silver chloride is observed. We have confirmed this by cell culture experiments (human mesenchymal stem cells; T-cells; monocytes) and bacteria (S. aureus) where the cells were incubated with synthetically prepared silver chloride nanoparticles (diameter ca. 100 nm). These were easily taken up by eukaryotic cells and showed the same toxic effect at the same silver concentration as ionic silver (as silver acetate). Therefore, nanoscopic silver chloride and not free ionic silver is the primary toxic species in biological media.

Critical influence of chloride ions on silver ion-mediated acute toxicity of silver nanoparticles to zebrafish embryos

The toxicity of silver nanoparticles (AgNP) to aquatic organisms, including zebrafish (Danio rerio), has been demonstrated, but differing opinions exist on the contribution of the physical properties of the particles themselves and the free dissolved silver ions (Ag+) to the observed effects. High concentrations of chloride ions (Cl−) in the routinely used exposure media can cause precipitation of Ag+ as AgCl, as well as complexation of silver in diverse soluble chlorocomplexes, thus masking the contribution of dissolved silver to AgNP toxicity. In the present study, we formulated a zebrafish exposure medium with a low chloride content and exposed zebrafish embryos to AgNO3 or carbonate-coated AgNP. The severity of toxicity caused by both silver forms depended on the time of exposure start, with younger embryos being most sensitive. Toxicity caused by both AgNO3 and AgNP was of the same order of magnitude when compared based on the total dissolved silver concentration and could be prevented by addition of the Ag+ chelator cysteine. Further, we have analyzed the data from several previous studies to evaluate the influence of interactions between Ag+ and Cl− on silver toxicity to zebrafish embryos. Our analysis demonstrates that the acute toxicity of AgNP to zebrafish embryos is largely mediated by Ag+. The influence of particle size and coating can at least partially be explained by the differences in Ag+ dissolution. High Cl− levels in the exposure medium indeed have a pivotal influence on the resulting toxicity of AgNP, appearing to significantly attenuate toxicity in several studies. This consideration should influence the choice of exposure medium to be used when evaluating and comparing AgNP toxicity.