Size- and Shape-Dependent Interactions of Lipid-Coated Silver Nanoparticles: An Improved Mechanistic Understanding through Model Cell Membranes and In Vivo Toxicity.

The widespread use of silver nanoparticles (AgNPs) in various applications and industries has brought to light the need for understanding the complex relationship between the physicochemical properties (shape, size, charge, and surface chemistry) of AgNPs that affect their ability to enter cells and cause toxicity. To evaluate their toxicological outcomes, this study systematically analyzed a series of homogeneous hybrid lipid-coated AgNPs spanning sizes from 5 to 100 nm with diverse shapes (spheres, triangles, and cubes). The hybrid lipid membrane comprises hydrogenated phosphatidylcholine (HPC), sodium oleate (SOA), and hexanethiol (HT), which shield the AgNP surface from surface oxidation and toxic Ag+ ion release to minimize its contribution to toxicity. To reduce any significant effects by surface chemistry, the HPC, SOA, and HT membrane composition ratio was kept constant, and the AgNPs were assessed using embryonic zebrafish (Danio rerio). While a direct comparison cannot be drawn due to the lack of complementary sizes below 40 nm for triangular plates and cubes due to synthetic challenges, significant mortality was observed for spherical AgNPs (AgNSs) of 5, 20, 40, and 60 nm at 120 h postfertilization at concentrations ≥6 mg Ag/L. In contrast, the 10, 80, and 100 nm AgNSs, 40, 70, and 100 nm triangular plate AgNPs (AgNPLs), and 55, 75, and 100 nm cubic AgNPs (AgNCs) showed no significant mortality at 5 days postfertilization following exposure to AgNPs at concentrations up to 12 mg Ag/L. With constant surface chemistry on the AgNPs, size is the dominant factor driving toxicological responses, with smaller nanoparticles (5 to 60 nm) being the most toxic. Larger AgNSs, AgNCs, and AgNPLs from 75 to 100 nm do not show any evidence of toxicity. However, when closely examining sizes between 40 and 60 nm for AgNSs, AgNCs, and AgNPLs, there is evidence that discriminates shape as a driver of toxicity since sublethal responses generally were observed to follow a pattern, suggesting toxicity is most significant for AgNSs followed by AgNPLs and then AgNCs, which is the least toxic. Sum frequency generation vibrational spectroscopy showed that irrespective of size or shape, all hybrid lipid-coated AgNPs interact with membrane surfaces and "snorkel" between phases into the lipid monolayer with minimal energetic cost. These findings decisively demonstrate that not only smaller AgNPs but also the shape of the AgNPs influences their biological compatibility.

Impacts of Differentially Shaped Silver Nanoparticles with Increasingly Complex Hydrophobic Thiol Surface Coatings in Small-Scale Laboratory Microcosms.

We investigated the impacts of spherical and triangular-plate-shaped lipid-coated silver nanoparticles (AgNPs) designed to prevent surface oxidation and silver ion (Ag+) dissolution in a small-scale microcosm to examine the role of shape and surface functionalization on biological interactions. Exposures were conducted in microcosms consisting of algae, bacteria, crustaceans, and fish embryos. Each microcosm was exposed to one of five surface chemistries within each shape profile (at 0, 0.1, or 0.5 mg Ag/L) to investigate the role of shape and surface composition on organismal uptake and toxicity. The hybrid lipid-coated AgNPs did not result in any significant release of Ag+ and had the most significant toxicity to D. magna, the most sensitive species, although the bacterial population growth rate was reduced in all exposures. Despite AgNPs resulting in increasing algal growth over the experiment, we found no correlation between algal growth and the survival of D. magna, suggesting that the impacts of the AgNPs on bacterial survival influenced algal growth rates. No significant impacts on zebrafish embryos were noted in any exposure. Our results demonstrate that the size, shape, and surface chemistry of AgNPs can be engineered to achieve specific goals while mitigating nanoparticle risks.

A Novel Approach for Identifying Nanoplastics by Assessing Deformation Behavior with Scanning Electron Microscopy.

As plastic production continues to increase globally, plastic waste accumulates and degrades into smaller plastic particles. Through chemical and biological processes, nanoscale plastic particles (nanoplastics) are formed and are expected to exist in quantities of several orders of magnitude greater than those found for microplastics. Due to their small size and low mass, nanoplastics remain challenging to detect in the environment using most standard analytical methods. The goal of this research is to adapt existing tools to address the analytical challenges posed by the identification of nanoplastics. Given the unique and well-documented properties of anthropogenic plastics, we hypothesized that nanoplastics could be differentiated by polymer type using spatiotemporal deformation data collected through irradiation with scanning electron microscopy (SEM). We selected polyvinyl chloride (PVC), polyethylene terephthalate (PET), and high-density polyethylene (HDPE) to capture a range of thermodynamic properties and molecular structures encompassed by commercially available plastics. Pristine samples of each polymer type were chosen and individually milled to generate micro and nanoscale particles for SEM analysis. To test the hypothesis that polymers could be differentiated from other constituents in complex samples, the polymers were compared against proxy materials common in environmental media, i.e., algae, kaolinite clay, and nanocellulose. Samples for SEM analysis were prepared uncoated to enable observation of polymer deformation under set electron beam parameters. For each sample type, particles approximately 1 µm in diameter were chosen, and videos of particle deformation were recorded and studied. Blinded samples were also prepared with mixtures of the aforementioned materials to test the viability of this method for identifying near-nanoscale plastic particles in environmental media. Based on the evidence collected, deformation patterns between plastic particles and particles present in common environmental media show significant differences. A computer vision algorithm was also developed and tested against manual measurements to improve the usefulness and efficiency of this method further.

Shape-dependent gold nanoparticle interactions with a model cell membrane.

Customizable gold nanoparticle platforms are motivating innovations in drug discovery with massive therapeutic potential due to their biocompatibility, stability, and imaging capabilities. Further development requires the understanding of how discrete differences in shape, charge, or surface chemistry affect the drug delivery process of the nanoparticle. The nanoparticle shape can have a significant impact on nanoparticle function as this can, for example, drastically change the surface area available for modifications, such as surface ligand density. In order to investigate the effects of nanoparticle shape on the structure of cell membranes, we directly probed nanoparticle-lipid interactions with an interface sensitive technique termed sum frequency generation (SFG) vibrational spectroscopy. Both gold nanostars and gold nanospheres with positively charged ligands were allowed to interact with a model cell membrane and changes in the membrane structure were directly observed by specific SFG vibrational modes related to molecular bonds within the lipids. The SFG results demonstrate that the +Au nanostars both penetrated and impacted the ordering of the lipids that made up the membrane, while very little structural changes to the model membrane were observed by SFG for the +Au nanospheres interacting with the model membrane. This suggests that the +Au nanostars, compared to the +Au nanospheres, are more disruptive to a cell membrane. Our findings indicate the importance of shape in nanomaterial design and provide strong evidence that shape does play a role in defining nanomaterial-biological interactions.

Silver Nanoparticles Stable to Oxidation and Silver Ion Release Show Size-Dependent Toxicity In Vivo.

Silver nanoparticles (AgNPs) are widely used in commerce, however, the effect of their physicochemical properties on toxicity remains debatable because of the confounding presence of Ag+ ions. Thus, we designed a series of AgNPs that are stable to surface oxidation and Ag+ ion release. AgNPs were coated with a hybrid lipid membrane comprised of L-phosphatidylcholine (PC), sodium oleate (SOA), and a stoichiometric amount of hexanethiol (HT) to produce oxidant-resistant AgNPs, Ag-SOA-PC-HT. The stability of 7-month aged, 20-100 nm Ag-SOA-PC-HT NPs were assessed using UV-Vis, dynamic light scattering (DLS), and inductively coupled plasma mass spectrometry (ICP-MS), while the toxicity of the nanomaterials was assessed using a well-established, 5-day embryonic zebrafish assay at concentrations ranging from 0-12 mg/L. There was no change in the size of the AgNPs from freshly made samples or 7-month aged samples and minimal Ag+ ion release (<0.2%) in fishwater (FW) up to seven days. Toxicity studies revealed AgNP size- and concentration-dependent effects. Increased mortality and sublethal morphological abnormalities were observed at higher concentrations with smaller nanoparticle sizes. This study, for the first time, determined the effect of AgNP size on toxicity in the absence of Ag+ ions as a confounding variable.

Effect of Nanoplastic Type and Surface Chemistry on Particle Agglomeration over a Salinity Gradient.

Agglomeration of nanoplastics in waters can alter their transport and fate in the environment. Agglomeration behavior of 4 nanoplastics differing in core composition (red- or blue-dyed polystyrene) and surface chemistry (plain or carboxylated poly[methyl methacrylate] [PMMA]) was investigated across a salinity gradient. No agglomeration was observed for carboxylated PMMA at any salinity, whereas the plain PMMA agglomerated at only 1 g/L. Both the red and the blue polystyrene agglomerated at 25 g/L. Results indicate that both composition and surface chemistry can impact how environmental salinity affects plastic nanoparticle agglomeration. Environ Toxicol Chem 2021;40:1822-1828. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

In Vivo Toxicity Assessment of Chitosan-Coated Lignin Nanoparticles in Embryonic Zebrafish (Danio rerio).

Lignin is the second most abundant biopolymer on Earth after cellulose. Since lignin breaks down in the environment naturally, lignin nanoparticles may serve as biodegradable carriers of biocidal actives with minimal environmental footprint compared to conventional antimicrobial formulations. Here, a lignin nanoparticle (LNP) coated with chitosan was engineered. Previous studies show both lignin and chitosan to exhibit antimicrobial properties. Another study showed that adding a chitosan coating can improve the adsorption of LNPs to biological samples by electrostatic adherence to oppositely charged surfaces. Our objective was to determine if these engineered particles would elicit toxicological responses, utilizing embryonic zebrafish toxicity assays. Zebrafish were exposed to nanoparticles with an intact chorionic membrane and with the chorion enzymatically removed to allow for direct contact of particles with the developing embryo. Both mortality and sublethal endpoints were analyzed. Mortality rates were significantly greater for chitosan-coated LNPs (Ch-LNPs) compared to plain LNPs and control groups. Significant sublethal endpoints were observed in groups exposed to Ch-LNPs with chorionic membranes intact. Our study indicated that engineered Ch-LNP formulations at high concentrations were more toxic than plain LNPs. Further study is warranted to fully understand the mechanisms of Ch-LNP toxicity.

Adaptive methodology to determine hydrophobicity of nanomaterials in situ.

The hydrophobicity of nanoparticles (NPs) is a key property determining environmental fate, biological partitioning and toxicity. However, methods to characterize surface hydrophobicity are not uniformly applied to NPs and cannot quantify surface changes in complex environments. Existing methods designed to evaluate the hydrophobicity of bulk solids, chemicals, and proteins have significant limitations when applied to NPs. In this study, we modified and evaluated two methods to determine the hydrophobicity of NPs, hydrophobic interaction chromatography (HIC) and dye adsorption, and compared them to the standard octanol-water partitioning protocol for chemicals. Gold, copper oxide, silica, and amine-functionalized silica NPs were used to evaluate methods based on their applicability to NPs that agglomerate and have surface coatings. The octanol water partitioning and HIC methods both measured Au NPs as hydrophilic, but despite having a small size and stable suspension, NPs could not be fully recovered from the HIC column. For the dye adsorption method, hydrophobic (Rose Bengal) and hydrophilic (Nile Blue) dyes were adsorbed to the NP surface, and linear isotherm parameters were used as a metric for hydrophobicity. CuO was determined to be slightly hydrophilic, while SiO2 was hydrophilic and Ami-SiO2 was hydrophobic. The advantages and limitations of each method are discussed, and the dye adsorption method is recommended as the most suitable for application across broad classes of nanomaterials. The dye assay method was further used to measure changes in the surface hydrophobicity of TiO2 NPs after being suspended in natural water collected from the Alsea Rivers watershed in Oregon. TiO2 NPs adsorbed Rose Bengal when suspended in ultrapure water, but adsorbed Nile Blue after being incubated in natural water samples, demonstrating a shift from hydrophobic to hydrophilic properties on the outer surface. The dye adsorption method can be applied to characterize surface hydrophobicity of NPs and quantify environmental transformations, potentially improving environmental fate models.

Assessment of Cu and CuO nanoparticle ecological responses using laboratory small-scale microcosms.

Copper based nanoparticles (NPs) are used extensively in industrial and commercial products as sensors, catalysts, surfactants, antimicrobials, and for other purposes. The high production volume and increasing use of copper-based NPs make their ecological risk a concern. Commonly used copper-based NPs are composed of metallic copper or copper oxide (Cu and CuO NPs); however, their environmental toxicity can vary dramatically depending on their physico-chemical properties, such as dissolution, aggregation behavior, and the generation of reactive oxygen species. Here, we investigated the NP dissolution, organismal uptake and aquatic toxicity of Cu and CuO NPs at 0, 0.1, 1, 5 or 10 mg Cu/L using a previously developed multi-species microcosm. This 5-day microcosm assay was comprised of C. reinhardtti, E. coli, D. magna, and D. rerio. We hypothesized that Cu and CuO NPs can elicit differential toxicity to the organisms due to alterations in particle dissolution and variations in organismal uptake. The actual concentrations of dissolved Cu released from the NPs were compared to ionic copper controls (CuCl2) at the same concentrations to determine the relative contribution of particulate and dissolved Cu on organism uptake and toxicity. We found that both NPs had higher uptake in D. magna and zebrafish than equivalent ionic exposures, suggesting that both Cu-based NPs are taken up by organisms. Cu NP exposures significantly inhibited algal growth rate, D. magna survival, and zebrafish hatching while exposure to equivalent concentrations of CuCl2 (dissolved Cu fraction) and CuO NPs did not. This indicates that Cu NPs themselves likely elicited a particle-specific mechanism of toxicity to the test organisms, or a combination effect from ionic Cu and the Cu NPs. Overall, this work was the first study to utilize a small-scale rapid assay designed to evaluate the fate and ecotoxicological impacts of Cu and CuO NPs in a mixed aquatic community.

Potential Risk to Pollinators from Nanotechnology-Based Pesticides.

The decline in populations of insect pollinators is a global concern. While multiple factors are implicated, there is uncertainty surrounding the contribution of certain groups of pesticides to losses in wild and managed bees. Nanotechnology-based pesticides (NBPs) are formulations based on multiple particle sizes and types. By packaging active ingredients in engineered particles, NBPs offer many benefits and novel functions, but may also exhibit different properties in the environment when compared with older pesticide formulations. These new properties raise questions about the environmental disposition and fate of NBPs and their exposure to pollinators. Pollinators such as honey bees have evolved structural adaptations to collect pollen, but also inadvertently gather other types of environmental particles which may accumulate in hive materials. Knowledge of the interaction between pollinators, NBPs, and other types of particles is needed to better understand their exposure to pesticides, and essential for characterizing risk from diverse environmental contaminants. The present review discusses the properties, benefits and types of nanotechnology-based pesticides, the propensity of bees to collect such particles and potential impacts on bee pollinators.

Monoalkyl Tin Nano-Cluster Films Reveal a Low Environmental Impact under Simulated Natural Conditions.

Recently, monoalkyl oxo-hydroxo tin clusters have emerged as a new class of metal-oxide resist to support the semiconductor industry's transition to extreme ultraviolet (EUV) lithography. Under EUV exposure, these tin-based clusters exhibit higher performance and wider process windows than conventional polymer materials. A promising new monoalkyl precursor, [(BuSn)12 O14 (OH)6 ][OH]2 (BuSn), is still in its infancy in terms of film formation. However, understanding potential environmental effects could significantly affect future development as a commercial product. We synthesized and explored the toxicity of nano-BuSn in the alga Chlamydomonas reinhardtii and the crustacean Daphnia magna at exposure concentrations ranging from 0 to 250 mg/L. Nano-BuSn had no effect on C. reinhardtii growth rate irrespective of concentration, whereas high nanoparticle concentrations (≥100 mg/L) increased D. magna immobilization and mortality significantly. To simulate an end-of-life disposal and leachate contamination, BuSn-coated film wafers were incubated in water at various pH values and temperatures for 14 and 90 d to investigate leaching rates and subsequent toxicity of the leachates. Although small quantities of tin (1.1-3.4% of deposited mass) leached from the wafers, it was insufficient to elicit a toxic response regardless of pH, incubation time, or temperature. The low toxicity of the tin-based thin films suggests that they can be an environmentally friendly addition to the material sets useful for semiconductor manufacturing. Environ Toxicol Chem 2019;38:2651-2658. © 2019 SETAC.

Comparative dissolution, uptake, and toxicity of zinc oxide particles in individual aquatic species and mixed populations.

Potential differences in species susceptibility to nanoparticle (NP) contaminants make the use of multispecies community toxicity testing strategies beneficial in understanding NP risk to aquatic environments. Because of the limited knowledge of zinc oxide (ZnO) NP fate and toxicity, we conducted multispecies exposures and compared the responses of individual species to the same species in a community comprised of algae (Chlamydomonas reinhardtii), bacteria (Escherichia coli), crustaceans (Daphnia magna), and zebrafish (Danio rerio). Different-sized ZnO particles and ionic Zn were compared to investigate the contribution of particulate and dissolved Zn to aquatic organism toxicity. Each organism and community was exposed to Zn sources at 0.08, 0.8, and 8 mg Zn/L. The present results indicate that all 3 types of Zn elicited differential toxicity among test organisms, with stronger adverse outcomes observed in single species than within a community. The community assay (nanocosm) we developed increased resilience to all Zn exposures by 5 to 10% compared to individual exposures at equivalent concentrations. In addition, the uptake and toxicity of ZnO particles to aquatic communities appear to be driven by rapid dissolution and the concomitant impacts of zinc ion toxicity, and the size of the ZnO particles had little impact on uptake or toxicity. The nanocosm assay could be a useful screening tool for rapidly assessing the potential impacts of nanomaterials to aquatic species. Environ Toxicol Chem 2019;38:591-602. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Reactive oxygen species generation is likely a driver of copper based nanomaterial toxicity.

Determining the specific nanomaterial features that elicit adverse biological responses is important to inform risk assessments, develop targeted applications, and rationally design future nanomaterials. Embryonic zebrafish are often employed to study nanomaterial-biological interactions, but few studies address the role of the chorion in nanomaterial exposure and toxicity. Here, we used chorion-intact (CI) or dechorionated (DC) embryonic zebrafish to investigate the influence of the chorion on copper-based nanoparticle toxicity. We found that despite higher dissolution and uptake, CuO NPs were less toxic than Cu NPs regardless of chorion status and did not cause 100 % mortality at even the highest exposure concentration. The presence of the chorion inhibited Cu toxicity: DC exposures to Cu NPs had an LC50 of 2.5 ± 0.3 mg/L compared to a CI LC50 of 13.7 ± 0.8 mg/L. This highlights the importance of considering zebrafish chorion status during nanotoxicological investigations, as embryo sensitivity increased by one order of magnitude or more when chorions were removed. Agglomerate size, zeta potential, and dissolved Cu did not sufficiently explain the differences in toxicity between Cu NPs and CuO NPs; however, reactive oxygen species (ROS) generation did. Cu NPs generated ROS in a concentration-dependent manner, while CuO did not and generated less than Cu NPs. We believe that the differences between the toxicities of Cu NPs and CuO NPs are due in part to their ability to generate ROS which could and should be a hazard consideration for risk assessments.

Toxicological Assessment of a Lignin Core Nanoparticle Doped with Silver as an Alternative to Conventional Silver Core Nanoparticles.

Elevated levels of silver in the environment are anticipated with an increase in silver nanoparticle (AgNP) production and use in consumer products. To potentially reduce the burden of silver ion release from conventional solid core AgNPs, a lignin-core particle doped with silver ions and surface-stabilized with a polycationic electrolyte layer was engineered. Our objective was to determine whether any of the formulation components elicit toxicological responses using embryonic zebrafish. Ionic silver and free surface stabilizer were the most toxic constituents, although when associated separately or together with the lignin core particles, the toxicity of the formulations decreased significantly. The overall toxicity of lignin formulations containing silver was similar to other studies on a silver mass basis, and led to a significantly higher prevalence of uninflated swim bladder and yolk sac edema. Comparative analysis of dialyzed samples which had leached their loosely bound Ag⁺, showed a significant increase in mortality immediately after dialysis, in addition to eliciting significant increases in types of sublethal responses relative to the freshly prepared non-dialyzed samples. ICP-OES/MS analysis indicated that silver ion release from the particle into solution was continuous, and the rate of release differed when the surface stabilizer was not present. Overall, our study indicates that the lignin core is an effective alternative to conventional solid core AgNPs for potentially reducing the burden of silver released into the environment from a variety of consumer products.

Uptake and toxicity of CuO nanoparticles to Daphnia magna varies between indirect dietary and direct waterborne exposures.

Research examining the direct and indirect ecological effects of nanomaterials in aquatic ecosystems is important for developing a more realistic understanding of the environmental implications of nanotechnology. Copper oxide nanoparticles (CuO NPs) are being used extensively in many industries but are considered highly toxic to aquatic species residing in surface waters. Few studies have addressed whether CuO NPs can be transferred through the aquatic food chain, and if such indirect exposure to nanomaterials impacts their toxicity. We investigated the uptake and trophic transfer of CuO NPs from the algae Chlorella vulgaris to the crustacean Daphnia magna and assessed bio-partitioning and resulting toxicity. We hypothesized that CuO NPs can be associated with algal cells and be transported to predators through feeding, and that the chronic toxicity can be altered in comparison to direct CuO NP exposure. For the indirect feeding exposure, algae pre-incubated with CuO NPs (Cu-algae) were washed to remove loose NPs and fed to D. magna while Cu uptake and toxicity were evaluated. For the direct waterborne exposures, a parallel group of D. magna were exposed to equivalent concentrations of CuO NPs while being fed unexposed algae. Using hyperspectral imaging we observed strong surface associations between pre-incubated CuO NPs and algae used in the feeding exposure, and quantified the average Cu content (0.15mg Cu/L) with ICP-OES. Cu accumulated in daphnid bodies to a greater extent in direct exposures, whereas molted carapaces and neonate offspring had more copper following the indirect feeding exposure, implying that D. magna may regulate internal Cu differently depending on the method of CuO NP delivery. Significantly higher D. magna mortality was observed following direct exposure relative to feeding exposure, and neonate production from adult daphnids exposed indirectly to CuO NPs was significantly reduced. Thus, nanoparticle interaction with biota at one trophic level may alter the biological response at the next trophic level in a way that is dependent on the delivery scenario. This study highlights the importance of evaluating potential ecological impacts of nanomaterials in more relevant, complex exposure scenarios.

Impacts of chemical modification on the toxicity of diverse nanocellulose materials to developing zebrafish.

Cellulose is an abundant and renewable resource currently being investigated for utility in nanomaterial form for various promising applications ranging from medical and pharmaceutical uses to mechanical reinforcement and biofuels. The utility of nanocellulose and wide implementation ensures increasing exposure to humans and the environment as nanocellulose-based technologies advance. Here, we investigate how differences in aspect ratio and changes to surface chemistry, as well as synthesis methods, influence the biocompatibility of nanocellulose materials using the embryonic zebrafish. Investigations into the toxicity of neutral, cationic and anionic surface functionalities revealed that surface chemistry had a minimal influence on the overall toxicity of nanocellulose materials. Higher aspect ratio cellulose nanofibers produced by mechanical homogenization were, in some cases, more toxic than other cellulose-based nanofibers or nanocrystals produced by chemical synthesis methods. Using fluorescently labeled nanocellulose we were able to show that nanocellulose uptake did occur in embryonic zebrafish during development. We conclude that the benign nature of nanocellulose materials makes them an ideal platform to systematically investigate the inherent surface features driving nanomaterial toxicity in order to create safer design principles for engineered nanoparticles.

The impact of aminated surface ligands and silica shells on the stability, uptake, and toxicity of engineered silver nanoparticles.

Inherent nanomaterial characteristics, composition, surface chemistry, and primary particle size, are known to impact particle stability, uptake, and toxicity. Nanocomposites challenge our ability to predict nanoparticle reactivity in biological systems if they are composed of materials with contrasting relative toxicities. We hypothesized that toxicity would be dominated by the nanoparticle surface (shell vs core), and that modulating the surface ligands would have a direct impact on uptake. We exposed developing zebrafish (Danio rerio) to a series of ~70 nm amine-terminated silver nanoparticles with silica shells (AgSi NPs) to investigate the relative influence of surface amination, composition, and size on toxicity. Like-sized aminated AgSi and Si NPs were more toxic than paired hydroxyl-terminated nanoparticles; however, both AgSi NPs were more toxic than the Si NPs, indicating a significant contribution of the silver core to the toxicity. Incremental increases in surface amination did not linearly increase uptake and toxicity, but did have a marked impact on dispersion stability. Mass-based exposure metrics initially supported the hypothesis that smaller nanoparticles (20 nm) would be more toxic than larger particles (70 nm). However, surface area-based metrics revealed that toxicity was independent of size. Our studies suggest that nanoparticle surfaces play a critical role in the uptake and toxicity of AgSi NPs, while the impact of size may be a function of the exposure metric used. Overall, uptake and toxicity can be dramatically altered by small changes in surface functionalization or exposure media. Only after understanding the magnitude of these changes, can we begin to understand the biologically available dose following nanoparticle exposure.

Comparative toxicological assessment of PAMAM and thiophosphoryl dendrimers using embryonic zebrafish.

Dendrimers are well-defined, polymeric nanomaterials currently being investigated for biomedical applications such as medical imaging, gene therapy, and tissue targeted therapy. Initially, higher generation (size) dendrimers were of interest because of their drug carrying capacity. However, increased generation was associated with increased toxicity. The majority of studies exploring dendrimer toxicity have focused on a small range of materials using cell culture methods, with few studies investigating the toxicity across a wide range of materials in vivo. The objective of the present study was to investigate the role of surface charge and generation in dendrimer toxicity using embryonic zebrafish (Danio rerio) as a model vertebrate. Due to the generational and charge effects observed at the cellular level, higher generation cationic dendrimers were hypothesized to be more toxic than lower generation anionic or neutral dendrimers with the same core composition. Polyamidoamine (PAMAM) dendrimers elicited significant morbidity and mortality as generation was decreased. No significant adverse effects were observed from the suite of thiophosphoryl dendrimers studied. Exposure to ≥50 ppm cationic PAMAM dendrimers G3-amine, G4-amine, G5-amine, and G6-amine caused 100% mortality by 24 hours post-fertilization. Cationic PAMAM G6-amine at 250 ppm was found to be statistically more toxic than both neutral PAMAM G6-amidoethanol and anionic PAMAM G6-succinamic acid at the same concentration. The toxicity observed within the suite of varying dendrimers provides evidence that surface charge may be the best indicator of dendrimer toxicity. Dendrimer class and generation are other potential contributors to the toxicity of dendrimers. Further studies are required to better understand the relative role each plays in driving the toxicity of dendrimers. To the best of our knowledge, this is the first in vivo study to address such a broad range of dendrimers.

Exposure assessment: recommendations for nanotechnology-based pesticides.

Given the development of nanotechnology within numerous scientific disciplines, it is likely that nanoscale products have been and will be used for agricultural, vector, and urban pest control prior to a complete evaluation of exposure and risk. Significant differences may exist between nanotechnology-based pesticides (NBPs) and conventional pesticides, primarily due to size and surface characteristics. These differences may result in changes in bioavailability, sensitivity, dosimetry, and pharmacokinetics. This paper considers the role of exposure assessment in the regulation of NBPs. While the existing regulatory infrastructure for pesticides is well established, several issues specific to NBP exposure are discussed, including: (1) disclosures of nanoparticle characteristics in product formulations; (2) additional uncertainty factors for NBPs with inadequate data; (3) route-specific approaches for assessing exposure; (4) testing with the commercial form of NBPs; (5) initiation of a health surveillance program; and (6) development of educational programs.