There is a current debate on whether the toxicity of engineered ZnO nanoparticles (NPs) can be traced back to their nanoscale properties or rather to the simple fact of their relatively high solubility and consequent release of Zn2+ ions. In this work, the emerging electroanalytical technique AGNES (Absence of Gradients and Nernstian Equilibrium Stripping), which is specially designed to determine free metal ion concentration, is shown to be able to measure the Zn2+ concentration resulting from dissolution of ZnO nanoparticles dispersed in aqueous salt solutions. Three NP samples from different sources (having average primary particle diameters of 6, 20, and 71 nm) were tested and compared with bulk ZnO material. The enhanced solubility of the nanoparticles with decreasing primary radius allows for an estimation of the surface energy of 0.32 J/m2. AGNES also allows the study of the kinetics of Zn2+ release as a response to a change in the solution parameters (e.g., pH, ZnO concentration). A physicochemical model has been developed to account for the observed kinetic behavior. With this model, only one kinetic parameter is required to describe the time dependence of the free Zn2+ concentration in solution. Good agreement with this prediction is obtained when, starting from an equilibrated NP dispersion, the pH of the medium is lowered. Also, the independence of this parameter from pH, as expected from the model, is obtained at least in the pH range 7–9. When dissolution is studied by dispersing ZnO nanoparticles in the medium, the kinetic parameter initially decreases with time. This decrease can be interpreted as resulting from the increase of the radius of the clusters due to the agglomeration/aggregation phenomena (independently confirmed). For the larger assayed NPs (i.e., 20 and 71 nm), a sufficiently large pH increase leads to a metastable solubility state, suggesting formation of a hydroxide interfacial layer.

ZnO nanoparticles (NPs) are prone to dissolution, and uncertainty remains whether biological/cellular responses to ZnO NPs are solely due to the release of Zn(2+) or whether the NPs themselves have additional toxic effects. We address this by establishing ZnO NP solubility in dispersion media (Dulbecco's modified Eagle's medium, DMEM) held under conditions identical to those employed for cell culture (37 °C, 5% CO2, and pH 7.68) and by systematic comparison of cell-NP interaction for three different ZnO NP preparations. For NPs at concentrations up to 5.5 μg ZnO/mL, dissolution is complete (with the majority of the soluble zinc complexed to dissolved ligands in the medium), taking ca. 1 h for uncoated and ca. 6 h for polymer coated ones. Above 5.5 μg/mL, the results are consistent with the formation of zinc carbonate, keeping the solubilized zinc fixed to 67 μM of which only 0.45 μM is as free Zn(2+), i.e., not complexed to dissolved ligands. At these relatively high concentrations, NPs with an aliphatic polyether-coating show slower dissolution (i.e., slower free Zn(2+) release) and reprecipitation kinetics compared to those of uncoated NPs, requiring more than 48 h to reach thermodynamic equilibrium. Cytotoxicity (MTT) and DNA damage (Comet) assay dose-response curves for three epithelial cell lines suggest that dissolution and reprecipitation dominate for uncoated ZnO NPs. Transmission electron microscopy combined with the monitoring of intracellular Zn(2+) concentrations and ZnO-NP interactions with model lipid membranes indicate that an aliphatic polyether coat on ZnO NPs increases cellular uptake, enhancing toxicity by enabling intracellular dissolution and release of Zn(2+). Similarly, we demonstrate that needle-like NP morphologies enhance toxicity by apparently frustrating cellular uptake. To limit toxicity, ZnO NPs with nonacicular morphologies and coatings that only weakly interact with cellular membranes are recommended.

In this study, the effect of ZnO nanoparticles and ZnCl2 on growth, reproduction and accumulation of zinc in Daphnia magna was determined in a 21-day chronic toxicity test. A variety of techniques were used to distinguish the free zinc ion, dissolved, nanoparticle and aggregated zinc fraction in the Daphnia test medium. The results showed similar chronic effects on growth, reproduction and accumulation for the ZnO nanoparticles (EC10, 20, 50 reproduction: 0.030, 0.049, 0.112 mg Zn/l) and the ZnCl2 (EC10, 20, 50 reproduction: 0.014, 0.027, 0.082 mg Zn/l). A large fraction of the nanoparticles rapidly dissolved after introduction in the exposure medium. Aggregation of nanoparticles was also observed but within 48 h of exposure most of these ZnO aggregates were dissolved. Based on the combined dissolution kinetics and toxicity results, it can be concluded that the toxicological effects of ZnO nanoparticles at the chronic level can be largely attributed to the dissolved fraction rather than the nanoparticles or initially formed aggregates.

We investigate the effect of serum albumin on the interaction of ZnO nanoparticles with DOPC lipid membranes and show that the size-stabilizing effect of the protein corona enhances their interaction with lipid membranes, which manifests, in part, as an increased ordering in the lipid packing.

This paper reports detailed characterisation of a zinc oxide (ZnO) nanopowder synthesized by a flame spray pyrolysis method. Detailed characterisation of the powder was carried out following a protocol that aims to determine key physicochemical characteristics that may affect its toxicity. Analysis by X-ray diffraction, (XRD), transmission electron microscopy (TEM) and surface area measurements confirmed monophasic hexagonal wurtzite ZnO nanoparticles with a specific surface area of 59 m 2 /g. Histograms derived from TEM analysis are presented to illustrate the polydispersity within the sample; particles were elongated in the c-crystallographic direction, with average length ~ 23 nm and width ~14 nm.Dynamic light scattering (0.1 w/v % in deionised water, pH 7.4) revealed the particles were agglomerated with a modal secondary particle size of ~ 1.5 μm. Fourier transform infra-red spectroscopy and X-ray photoelectron spectroscopy indicated the presence of carbonate impurities on the surface of the ZnO nanoparticles.