Dik van de Meent

Cellular uptake of nanoparticles as determined by particle properties, experimental conditions, and cell type

The increased application of nanoparticles (NPs) is increasing the risk of their release into the environment. Although many toxicity studies have been conducted, the environmental risk is difficult to estimate, because uptake mechanisms are often not determined in toxicity studies. In the present study, the authors review dominant uptake mechanisms of NPs in cells, as well as the effect of NP properties, experimental conditions, and cell type on NP uptake. Knowledge of NP uptake is crucial for risk assessment and is essential to predict the behavior of NPs based on their physical-chemical properties. Important uptake mechanisms for eukaryotic cells are macropinocytosis, receptor-mediated endocytosis, and phagocytosis in specialized mammalian cells. The studies reviewed demonstrate that uptake into nonphagocytic cells depends strongly on NP size, with an uptake optimum at an NP diameter of approximately 50 nm. Increasing surface charges, either positive or negative, have been shown to increase particle uptake in comparison with uncharged NPs. Another important factor is the degree of (homo-) aggregation. Results regarding shape have been ambiguous. Difficulties in the production of NPs, with 1 property changed at a time, call for a full characterization of NP properties. Only then will it be possible to draw conclusions as to which property affected the uptake.

Aquatic ecotoxicity tests of some nanomaterials

Nanoparticles of TiO2, ZrO2, AL2O3, CeO2, fullerene (C60), single-walled carbon nanotubes, and polymethylmethacrylate were tested for ecotoxic effects using one or more ecotoxicity endpoints: Microtox (bacteria), pulse-amplitude modulation (algae), Chydotox (crustaceans), and Biolog (soil enzymes). No appreciable effects were observed at nominal concentrations of up to 100 mg/L. Dilution of nanoparticle suspensions, either in ultrapure (Milli-Q) water or in natural (pond) water, led to formation of larger particles, which settled easily. (Nano)particles in water were characterized by means of atomic force microscopy, energy-dispersive x-ray analysis, inductively coupled plasma-mass spectrometry, flow cytometry, and spectrophotometry. It is concluded that the absence of ecotoxicity is the result of low concentrations of free nanoparticles in the tests, and it is suggested that colloid (in)stability is of primary importance in explaining ecotoxic effects of nanoparticles in the natural environment.

Effect of natural organic matter on cerium dioxide nanoparticles settling in model fresh water.

The ecological risk assessment of chemicals including nanoparticles is based on the determination of adverse effects on organisms and on the environmental concentrations to which biota are exposed. The aim of this work was to better understand the behavior of nanoparticles in the environment, with the ultimate goal of predicting future exposure concentrations in water. We measured the concentrations and particle size distributions of CeO(2) nanoparticles in algae growth medium and deionized water in the presence of various concentrations and two types of natural organic matter (NOM). The presence of natural organic matter stabilizes the CeO(2) nanoparticles in suspension. In presence of NOM, up to 88% of the initially added CeO(2) nanoparticles remained suspended in deionized water and 41% in algae growth medium after 12d of settling. The adsorbed organic matter decreases the zeta potential from about -15 mV to -55 mV. This reduces aggregation by increased electrostatic repulsion. The particle diameter, pH, electric conductivity and NOM content shows significant correlation with the fraction of CeO(2) nanoparticles remaining in suspension.

Human-Toxicological Effect and Damage Factors of Carcinogenic and Noncarcinogenic Chemicals for Life Cycle Impact Assessment

Abstract Chemical fate, effect, and damage should be accounted for in the analysis of human health impacts by toxic chemicals in life-cycle assessment (LCA). The goal of this article is to present a new method to derive human damage and effect factors of toxic pollutants, starting from a lognormal dose–response function. Human damage factors are expressed as disability-adjusted life years (DALYs). Human effect factors contain a disease-specific and a substance-specific component. The disease-specific component depends on the probability of disease occurrence and the distribution of sensitivities in the human population. The substance-specific component, equal to the inverse of the ED50, represents the toxic potency of a substance. The new method has been applied to calculate combined human damage and effect factors for 1,192 substances. The total range of 7 to 9 orders of magnitude between the substances is dominated by the range in toxic potencies. For the combined factors, the typical uncertainty, represented by the square root of the ratio of the 97.5th and 2.5th percentile, is a factor of 25 for carcinogenic effects and a factor of 125 for noncarcinogenic effects. The interspecies conversion factor, the (non)cancer effect conversion factor, and the average noncancer damage factor dominate the overall uncertainty.

Natural colloids are the dominant factor in the sedimentation of nanoparticles

Estimating the environmental exposure to manufactured nanomaterials is part of risk assessment. Because nanoparticles aggregate with each other (homoaggregation) and with other particles (heteroaggregation), the main route of the removal of most nanoparticles from water is aggregation, followed by sedimentation. The authors used water samples from two rivers in Europe, the Rhine and the Meuse. To distinguish between small (mainly natural organic matter [NOM]) particles and the remainder of the natural colloids present, both filtered and unfiltered river water was used to prepare the particle suspensions. The results show that the removal of nanoparticles from natural river water follows first-order kinetics toward a residual concentration. This was measured in river water with less than 1 mg L(-1) CeO(2) nanoparticles. The authors inferred that the heteroaggregation with or deposition onto the solid fraction of natural colloids was the main mechanism causing sedimentation in relation to homoaggregation. In contrast, the NOM fraction in filtered river water stabilized the residual nanoparticles against further sedimentation for up to 12 d. In 10 mg L(-1) and 100 mg L(-1) CeO(2) nanoparticle suspensions, homoaggregation is likely the main mechanism leading to sedimentation. The proposed model could form the basis for improved exposure assessment for nanomaterials.

Multimedia modeling of engineered nanoparticles with simplebox4nano: Model definition and evaluation

Screening level models for environmental assessment of engineered nanoparticles (ENP) are not generally available. Here, we present SimpleBox4Nano (SB4N) as the first model of this type, assess its validity, and evaluate it by comparisons with a known material flow model. SB4N expresses ENP transport and concentrations in and across air, rain, surface waters, soil, and sediment, accounting for nanospecific processes such as aggregation, attachment, and dissolution. The model solves simultaneous mass balance equations (MBE) using simple matrix algebra. The MBEs link all concentrations and transfer processes using first-order rate constants for all processes known to be relevant for ENPs. The first-order rate constants are obtained from the literature. The output of SB4N is mass concentrations of ENPs as free dispersive species, heteroaggregates with natural colloids, and larger natural particles in each compartment in time and at steady state. Known scenario studies for Switzerland were used to demonstrate the impact of the transport processes included in SB4N on the prediction of environmental concentrations. We argue that SB4N-predicted environmental concentrations are useful as background concentrations in environmental risk assessment.

How to assess exposure of aquatic organisms to manufactured nanoparticles

Ecological risk of chemicals is measured by the quotient of predicted no-effect concentrations and predicted exposure concentrations, which are hard to assess for manufactured nanomaterials (NMs). This paper proposes modifications to currently used models, in order to make them suitable for estimating exposure concentrations of NMs in the aquatic environment. We have evaluated the adequacy of the current guidance documents for use with NMs and conclude that nano-specific fate processes, such as sedimentation and dissolution need to be incorporated. We have reviewed the literature on sedimentation and dissolution of NMs in environmentally relevant systems. We deduce that the overall kinetics of water-sediment transport of NMs should be close to first order. The lack of data on dissolution of NMs under environmentally realistic conditions calls for a pragmatic decision on which rates to be used in modeling. We find that first order removal kinetics for dissolution seems adequate. Based on limited data from literature, probable removal rates range from 0 to 10(-4)s(-1) for sedimentation, and from 0 to 10(-5)s(-1) for dissolution. Further experimental data at environmentally relevant conditions for sedimentation and dissolution of NMs is needed.

The contribution of complexed copper to the metabolic inhibition of algae and bacteria in synthetic media and river water

The toxicity of copper is reduced due to complexation by ligands, e.g. humic acids or synthetic compounds like EDTA. Therefore, the concentration of free metal ions has been considered the main determinant of metal toxicity. This early hypothesis was tested by adding different concentrations of copper to water from the River Rhine and to a synthetic medium, supplemented with different concentrations of EDTA. Subsequently the following parameters were determined: (1) voltammetrically labile copper (differential pulse anodic stripping voltammetry), (2) fraction of copper retained on Chelex-100 columns, (3) photosynthetic rate of the alga Selenastrum capricornutum (in synthetic medium) and (4) rate of [3H]thymidine incorporation by multiplying bacteria (in river water). Addition of 5μM Cu to the medium with 5 or 10μM of EDTA inhibited algal photosynthesis, although copper was not voltammetrically detectable (<0.005 μM). Small spikes of copper (0.06-1.00μM) added to river water inhibited the multiplication of bacteria; electrochemical determination showed no detectable copper activity at the lower concentrations. Chelex-retained copper correlated well with the percentage inhibition of the two biological activities. According to calculations using the model TITRATOR copper-EDTA complexes and un-ionized salts (mainly CuCO3) were dominant copper species in synthetic solutions inhibiting photosynthesis. The GECHEQ model calculated that free copper as well as chelex-labile copper correlated well with the inhibition of bacterial growth rate. Therefore it seems that complexed copper is “biologically available” to a significant extent. Furthermore, intermediate labile copper, such as chelex-retained copper, is an appropriate measure of copper toxicity.

Rapid settling of nanoparticles due to heteroaggregation with suspended sediment

Sedimentation of engineered nanoparticles (ENPs) has been studied mainly in artificial media and stagnant systems mimicking natural waters. This neglects the role of turbulence and heteroaggregation with sediment. The authors studied the apparent sedimentation rates of selected ENPs (cerium dioxide [CeO2 ], polyvinylpyrrolidone-capped silver [PVP-Ag], and silica-coated silver [SiO2 -Ag]) in agitated sediment-water systems resembling fresh, estuarine, and marine waters. Experiments were designed to mimic low energy and periodically resuspended sediment water systems (14 d), followed by a long-term aging, resuspension, and settling phase (6 months), as would occur in receiving shallow lakes. The ENPs in systems with periodical resuspension of sediment were removed with sedimentation rates between 0.14 m/d and 0.50 m/d. The sedimentation rates did not vary much among ENP type, salinity, and aging time, which is attributed to the capture of ENPs in sediment flocks. The sedimentation rates were 1 to 2 orders of magnitude higher than those reported for aggregation-sedimentation in stagnant systems without suspended sediment. Heteroaggregation rates were estimated and ranged between 0.151 L/mg/d and 0.547 L/mg/d, which is up to 29 times higher than those reported for natural colloids under quiescent settling conditions. The authors conclude that rapid scavenging and sedimentation drives removal of ENPs from the water column.

Calculating life-cycle assessment effect factors from potentially affected fraction-based ecotoxicological response functions

A new multisubstance potentially affected fraction (msPAF)-based method for calculating ecotoxicological effect factors for life-cycle assessment is introduced and compared to two other available methods of calculation. The new method is based on marginal increase of the msPAF of species. The method follows concentration-additive rules for pollutants with the same toxic mode of action (TMoA) and response-additive calculation rules for pollutants with independent action, and it combines a TMoA-specific factor, which is calculated differently in different methods, and a substance-specific factor, which is common to all methods. For 261 substances in 22 toxic modes of action, ecotoxicological effect factors for freshwater ecosystems have been calculated by different methods. Method-related differences appear to be rather small. Intersubstance differences in effect factors stem from differences in substance-specific toxic potencies, which span eight orders of magnitude, rather than from differences in TMoA-specific factors, which span only three orders of magnitude. Based on these insights, the choice of a calculation method seems to be a matter of personal (scientific) preference. The new hybrid msPAF method was greatly sensitive to data that usually are not known with sufficient certainty. Its use is recommended, entirely for reasons of scientific consistency, under strict conditions. When such conditions are not met or if necessary parameter values are unavailable, use of a fixed value for the TMoA-specific component is recommended.