Exposure of silver-nanoparticles and silver-ions to lung cells in vitro at the air-liquid interface

BACKGROUND: Due to its antibacterial properties, silver (Ag) has been used in more consumer products than any other nanomaterial so far. Despite the promising advantages posed by using Ag-nanoparticles (NPs), their interaction with mammalian systems is currently not fully understood. An exposure route via inhalation is of primary concern for humans in an occupational setting. Aim of this study was therefore to investigate the potential adverse effects of aerosolised Ag-NPs using a human epithelial airway barrier model composed of A549, monocyte derived macrophage and dendritic cells cultured in vitro at the air-liquid interface. Cell cultures were exposed to 20 nm citrate-coated Ag-NPs with a deposition of 30 and 278 ng/cm2 respectively and incubated for 4 h and 24 h. To elucidate whether any effects of Ag-NPs are due to ionic effects, Ag-Nitrate (AgNO3) solutions were aerosolised at the same molecular mass concentrations. RESULTS: Agglomerates of Ag-NPs were detected at 24 h post exposure in vesicular structures inside cells but the cellular integrity was not impaired upon Ag-NP exposures. Minimal cytotoxicity, by measuring the release of lactate dehydrogenase, could only be detected following a higher concentrated AgNO3-solution. A release of pro-inflammatory markers TNF-alpha and IL-8 was neither observed upon Ag-NP and AgNO3 exposures as well as was not affected when cells were pre-stimulated with lipopolysaccharide (LPS). Also, an induction of mRNA expression of TNF-alpha and IL-8, could only be observed for the highest AgNO3 concentration alone or even significantly increased when pre-stimulated with LPS after 4 h. However, this effect disappeared after 24 h. Furthermore, oxidative stress markers (HMOX-1, SOD-1) were expressed after 4 h in a concentration dependent manner following AgNO3 exposures only. CONCLUSIONS: With an experimental setup reflecting physiological exposure conditions in the human lung more realistic, the present study indicates that Ag-NPs do not cause adverse effects and cells were only sensitive to high Ag-ion concentrations. Chronic exposure scenarios however, are needed to reveal further insight into the fate of Ag-NPs after deposition and cell interactions.

Bioavailability of silver nanoparticles and ions: from a chemical and biochemical perspective

Owing to their antimicrobial properties, silver nanoparticles (NPs) are the most commonly used engineered nanomaterial for use in a wide array of consumer and medical applications. Many discussions are currently ongoing as to whether or not exposure of silver NPs to the ecosystem (i.e. plants and animals) may be conceived as harmful or not. Metallic silver, if released into the environment, can undergo chemical and biochemical conversion which strongly influence its availability towards any biological system. During this process, in the presence of moisture, silver can be oxidized resulting in the release of silver ions. To date, it is still debatable as to whether any biological impact of nanosized silver is relative to either its size, or to its ionic constitution. The aim of this review therefore is to provide a comprehensive, interdisciplinary overview--for biologists, chemists, toxicologists as well as physicists--regarding the production of silver NPs, its (as well as in their ionic form) chemical and biochemical behaviours towards/within a multitude of relative and realistic biological environments and also how such interactions may be correlated across a plethora of different biological organisms.

Comparative Cytotoxicity Study of Silver Nanoparticles (AgNPs) in a Variety of Rainbow Trout Cell Lines (RTL-W1, RTH-149, RTG-2) and Primary Hepatocytes.

Among all classes of nanomaterials, silver nanoparticles (AgNPs) have potentially an important ecotoxicological impact, especially in freshwater environments. Fish are particularly susceptible to the toxic effects of silver ions and, with knowledge gaps regarding the contribution of dissolution and unique particle effects to AgNP toxicity, they represent a group of vulnerable organisms. Using cell lines (RTL-W1, RTH-149, RTG-2) and primary hepatocytes of rainbow trout (Oncorhynchus mykiss) as in vitro test systems, we assessed the cytotoxicity of the representative AgNP, NM-300K, and AgNO3 as an Ag+ ion source. Lack of AgNP interference with the cytotoxicity assays (AlamarBlue, CFDA-AM, NRU assay) and their simultaneous application point to the compatibility and usefulness of such a battery of assays. The RTH-149 and RTL-W1 liver cell lines exhibited similar sensitivity as primary hepatocytes towards AgNP toxicity. Leibovitz's L-15 culture medium composition (high amino acid content) had an important influence on the behaviour and toxicity of AgNPs towards the RTL-W1 cell line. The obtained results demonstrate that, with careful consideration, such an in vitro approach can provide valuable toxicological data to be used in an integrated testing strategy for NM-300K risk assessment.

Influence of thymol and silver nanoparticles on the degradation of poly(lactic acid) based nanocomposites: thermal and morphological properties

Biopolymers, such as poly(lactic acid) (PLA), have been proposed as environmentally-friendly alternatives in applications such as food packaging. In this work, silver nanoparticles and thymol were used as active additives in PLA matrices, combining the antibacterial activity of silver with the antioxidant performance of thymol. The combined action of both additives influenced PLA thermal degradation in ternary systems. DSC results showed that the addition of thymol resulted in a clear decrease of the glass transition temperature (Tg) of PLA, suggesting its plasticizing effect in PLA matrices. Slight modifications in mechanical properties of dog-bone bars were also observed after the addition of the active components, especially in the elastic modulus. FESEM analyses showed the good distribution of active additives through the PLA matrix, obtaining homogenous surfaces and highlighting the presence of silver nanoparticles successfully embedded into the bulk matrix. Degradation of these PLA-based nanocomposites with thymol and silver nanoparticles in composting conditions indicated that the inherent biodegradable character of this biopolymer was improved after this modification. The obtained nanocomposites showed suitable properties to be used as biodegradable active-food packaging systems with antioxidant and antimicrobial effects.

Nano-biocomposite films with modified cellulose nanocrystals and synthesized silver nanoparticles

Ternary nano-biocomposite films based on poly(lactic acid) (PLA) with modified cellulose nanocrystals (s-CNC) and synthesized silver nanoparticles (Ag) have been prepared and characterized. The functionalization of the CNC surface with an acid phosphate ester of ethoxylated nonylphenol favoured its dispersion in the PLA matrix. The positive effects of the addition of cellulose and silver on the PLA barrier properties were confirmed by reductions in the water permeability (WVP) and oxygen transmission rate (OTR) of the films tested. The migration level of all nano-biocomposites in contact with food simulants were below the permitted limits in both non-polar and polar simulants. PLA nano-biocomposites showed a significant antibacterial activity influenced by the Ag content, while composting tests showed that the materials were visibly disintegrated after 15 days with the ternary systems showing the highest rate of disintegration under composting conditions.

Combined effects of cellulose nanocrystals and silver nanoparticles on the barrier and migration properties of PLA nano-biocomposites

Poly(lactic acid) (PLA)-based high performance nano-biocomposites were prepared to be used in active food packaging. Pristine (CNC) and surfactant modified cellulose nanocrystals (s-CNC) with silver (Ag) nanoparticles were used as the matrix modifiers. Binary and ternary systems were prepared. Morphological investigations revealed the good distribution of silver nanoparticles in PLA ternary systems. The combination of s-CNC and Ag nanoparticles increased the barrier effect of the produced films while the results of overall migration for the PLA nano-biocomposites revealed that none of the samples exceeded the overall migration limit, since results were well below 60 mg kg−1 of simulant.

SILVER NANOPARTICLES AND THE PLANT-ASSOCIATING ABILITIES OF RHIZOBIACEAE BACTERIA

The use of nanoparticle technology in consumer products has been increasing due to their broad-spectrum antimicrobial properties. Specifically, silver nanoparticles (AgNPs) can demonstrate distinct physiochemical properties compared to bulk silver, including a large surface area to volume ratio that allows for higher reactivity with bacterial cell surfaces. AgNPs are being released into the environment, including soil ecosystems through various pathways such as points of production or during disposal of silver-containing products. This raises the concern about the potential impact on beneficial soil bacteria and their surrounding ecosystems. Members of the Rhizobiaceae family play important roles in nutrient cycling and contribute to overall soil fertility and the experiments in this thesis address the potential for AgNP-mediated toxicity on these plant-associating bacteria. Respiration analysis of Bradyrhizobium japonicum, Azospirillum brasilense, and Agrobacterium tumefaciens has revealed that AgNPs can negatively impact the growth and survival of these bacterial species, with B. japonicum being the most susceptible. Additionally, swimming motility assays using B. japonicum showed a significant decrease in colony diameter when treated with AgNPs (50 ppm). A significant decrease in root colonization of Triticum aestivum roots by A. brasilense was observed as AgNP treatment concentrations increased. Although some of the experiments could not be completed, taken together, these experiments and the research reported herein highlights the potential toxicological effects of AgNPs on bacterial species vital to the growth and health of agriculturally important crops.

RIR-MAPLE deposition of plasmonic silver nanoparticles

© 2016, Springer-Verlag Berlin Heidelberg.Nanoparticles are being explored in many different applications due to the unique properties offered by quantum effects. To broaden the scope of these applications, the deposition of nanoparticles onto substrates in a simple and controlled way is highly desired. In this study, we use resonant infrared matrix-assisted pulsed laser evaporation (RIR-MAPLE) for the deposition of metallic, silver nanoparticles for plasmonic applications. We find that RIR-MAPLE, a simple and versatile approach, is able to deposit silver nanoparticles as large as 80 nm onto different substrates with good adhesion, regardless of substrate properties. In addition, the nanoparticle surface coverage of the substrates, which result from the random distribution of nanoparticles across the substrate per laser pulse, can be simply and precisely controlled by RIR-MAPLE. Polymer films of poly(3-hexylthiophene-2,5-diyl) (P3HT) are also deposited by RIR-MAPLE on top of the deposited silver nanoparticles in order to demonstrate enhanced absorption due to the localized surface plasmon resonance effect. The reported features of RIR-MAPLE nanoparticle deposition indicate that this tool can enable efficient processing of nanoparticle thin films for applications that require specific substrates or configurations that are not easily achieved using solution-based approaches.

SILVER NANOPARTICLES AND THE PLANT-ASSOCIATING ABILITIES OF RHIZOBIACEAE BACTERIA

The use of nanoparticle technology in consumer products has been increasing due to their broad-spectrum antimicrobial properties. Specifically, silver nanoparticles (AgNPs) can demonstrate distinct physiochemical properties compared to bulk silver, including a large surface area to volume ratio that allows for higher reactivity with bacterial cell surfaces. AgNPs are being released into the environment, including soil ecosystems through various pathways such as points of production or during disposal of silver-containing products. This raises the concern about the potential impact on beneficial soil bacteria and their surrounding ecosystems. Members of the Rhizobiaceae family play important roles in nutrient cycling and contribute to overall soil fertility and the experiments in this thesis address the potential for AgNP-mediated toxicity on these plant-associating bacteria. Respiration analysis of Bradyrhizobium japonicum, Azospirillum brasilense, and Agrobacterium tumefaciens has revealed that AgNPs can negatively impact the growth and survival of these bacterial species, with B. japonicum being the most susceptible. Additionally, swimming motility assays using B. japonicum showed a significant decrease in colony diameter when treated with AgNPs (50 ppm). A significant decrease in root colonization of Triticum aestivum roots by A. brasilense was observed as AgNP treatment concentrations increased. Although some of the experiments could not be completed, taken together, these experiments and the research reported herein highlights the potential toxicological effects of AgNPs on bacterial species vital to the growth and health of agriculturally important crops.