Sulfidation of silver nanoparticles: natural antidote to their toxicity.

Nanomaterials are highly dynamic in biological and environmental media. A critical need for advancing environmental health and safety research for nanomaterials is to identify physical and chemical transformations that affect the nanomaterial properties and their toxicity. Silver nanoparticles, one of the most toxic and well-studied nanomaterials, readily react with sulfide to form Ag(0)/Ag2S core-shell particles. Here, we show that sulfidation decreased silver nanoparticle toxicity to four diverse types of aquatic and terrestrial eukaryotic organisms (Danio rerio (zebrafish), Fundulus heteroclitus (killifish), Caenorhabditis elegans (nematode worm), and the aquatic plant Lemna minuta (least duckweed)). Toxicity reduction, which was dramatic in killifish and duckweed even for low extents of sulfidation (about 2 mol % S), is primarily associated with a decrease in Ag(+) concentration after sulfidation due to the lower solubility of Ag2S relative to elemental Ag (Ag(0)). These results suggest that even partial sulfidation of AgNP will decrease the toxicity of AgNPs relative to their pristine counterparts. We also show that, for a given organism, the presence of chloride in the exposure media strongly affects the toxicity results by affecting Ag speciation. These results highlight the need to consider environmental transformations of NPs in assessing their toxicity to accurately portray their potential environmental risks.

Theoretical framework for nanoparticle reactivity as a function of aggregation state.

Theory is developed that relates the reactivity of nanoparticles to the structure of aggregates they may form in suspensions. This theory is applied to consider the case of reactive oxygen species (ROS) generation by photosensitization of C(60) fullerenes. Variations in aggregate structure and size appear to account for an apparent paradox in ROS generation as calculated using values for the photochemical kinetics of fullerene (C(60)) and its hydroxylated derivative, fullerol (C(60)(OH)(22-24)) and assuming that structure varies between compact and fractal objects. A region of aggregation-suppressed ROS production is identified where interactions between the particles in compact aggregates dominate the singlet oxygen production. Intrinsic kinetic properties dominate when aggregates are small and/or are characterized by low fractal dimensions. Pseudoglobal sensitivity analysis of model input variables verifies that fractal dimension, and by extension aggregation state, is the most sensitive model parameter when kinetics are well-known. This theoretical framework qualitatively predicts ROS production by fullerol suspensions 2 orders of magnitude higher compared with aggregates of largely undifferentiated C(60) despite nearly an order of magnitude higher quantum yield for the undifferentiated C(60) based on measurements for single molecules. Similar to C(60), other primary nanoparticles will exist as aggregates in many environmental and laboratory suspensions. This work provides a theoretical basis for understanding how the structure of nanoparticle aggregates may affect their reactivity.

Nanoparticle aggregation: challenges to understanding transport and reactivity in the environment.

Unique forms of manufactured nanomaterials, nanoparticles, and their suspensions are rapidly being created by manipulating properties such as shape, size, structure, and chemical composition and through incorporation of surface coatings. Although these properties make nanomaterial development interesting for new applications, they also challenge the ability of colloid science to understand nanoparticle aggregation in the environment and the subsequent effects on nanomaterial transport and reactivity. This review briefly covers aggregation theory focusing on Derjaguin-Landau-Verwey-Overbeak (DLVO)-based models most commonly used to describe the thermodynamic interactions between two particles in a suspension. A discussion of the challenges to DLVO posed by the properties of nanomaterials follows, along with examples from the literature. Examples from the literature highlighting the importance ofaggregation effects on transport and reactivity and risk of nanoparticles in the environment are discussed.

Environmental occurrences, behavior, fate, and ecological effects of nanomaterials: an introduction to the special series.

The release of engineered nanomaterials (ENMs) into the biosphere will increase as industries find new and useful ways to utilize these materials. Scientists and engineers are beginning to assess the material properties that determine the fate, transport, and effects of ENMs; however, the potential impacts of released ENMs on organisms, ecosystems, and human health remain largely unknown. This special collection of four review papers and four technical papers identifies many key and emerging knowledge gaps regarding the interactions between nanomaterials and ecosystems. These critical knowledge gaps include the form, route, and mass of nanomaterials entering the environment; the transformations and ultimate fate of nanomaterials in the environment; the transport, distribution, and bioavailability of nanomaterials in environmental media; and the organismal responses to nanomaterial exposure and effects of nanomaterial inputs, on ecological communities and biogeochemical processes at relevant environmental concentrations and forms. This introductory section summarizes the state of knowledge and emerging areas of research needs identified within the special collection. Despite recent progress in understanding the transport, transformations, and fate of ENMs in model environments and organisms, there remains a large need for fundamental information regarding releases, distribution, transformations and persistence, and bioavailability of nanomaterials. Moreover, fate, transport, bioaccumulation, and ecological impacts research is needed using environmentally relevant concentrations and forms of ENMs in real field materials and with a broader range of organisms.

Mechanisms of bacteriophage inactivation via singlet oxygen generation in UV illuminated fullerol suspensions.

Nonenveloped viruses are shown to be inactivated by singlet oxygen ((1)O2) produced in UVA photosensitized aqueous suspensions of a polyhydroxylated fullerene (C60(OH)22-24; fullerol, 40 microM). Experiments were performed with MS2, a ssRNA bacteriophage, as well as two dsDNA phages: PRD1, which has an internal lipid membrane, and T7, which entirely lacks lipids. MS2 was highly susceptible to inactivation, having a rate constant of 0.034 min(-1) with UVA alone, which increased to 0.102 min(-1) with photoactivated fullerol. PRD1 and T7 were not susceptible to UVA alone but were photoinactivated by fullerol with rate constants of 0.026 and 0.035 min(-1), respectively. The role of 1(O)2 was demonstrated by three independent observations: (i) viruses that were insensitive to UVA alone were photoinactivated by rose bengal in the absence of fullerol, (ii) beta-carotene reduced (but did not eliminate) photoinactivation rates, and (iii) singlet oxygen sensor green fluorescence spectroscopy directly detected (1)O2 in UVA illuminated fullerol suspensions. Qualitative evidence is also presented that fullerol aggregates were closely associated with viruses allowing efficient transfer of 1(O)2 to their capsids. Fourier transform infrared spectroscopy revealed significant oxidative modifications to capsid proteins but comparatively minor changes to the DNA and (phospho)lipids. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) suggested (1)O2 induced crosslinking of proteins. Hence, phage inactivation by photoactivated fullerol nanoparticles appears to be caused by cross-linking of capsid protein secondary structures by exogenous (1)O2 and consequentimpairmentof their ability to bind to surface receptors of their bacterial hosts (loss of infectivity) rather than by direct reactions with fullerol.

Evaluation of the oxidation of organic compounds by aqueous suspensions of photosensitized hydroxylated-C60 fullerene aggregates.

Ultraviolet (UV) irradiated polyhydroxylated fullerene (fullerol) nanomaterials are examined for their potential to degrade organic compounds via reactive oxygen species (ROS) mediated by a photosensitization process. Organic compounds were selected for their sensitivity to individual species of reactive oxygen (hydroxyl radical (*OH-) for degradation of salicylic acid (SA); singlet oxygen (1O2) for degradation of 2-chlorophenol (2CP), and superoxide (O2*-) for oxidation of ethanol) and were monitored over time in aqueous suspensions of fullerol aggregates. Only the 2CP showed significant degradation underscoring the specificity of the fullerol in producing singlet oxygen in these conditions. Monitoring these processes via high performance liquid chromatography (HPLC) confirmed that organic compounds degraded primarily by ROS over a range of fullerol concentrations, pH values, and temperatures.

Comparative toxicity of C60 aggregates toward mammalian cells: role of tetrahydrofuran (THF) decomposition.

C60 fullerene is a promising material because of its unique physiochemical properties. However, previous studies have reported that colloidal aggregates of C60 (nC60) produce toxicity in fish and human cell cultures. The preparation method of nC60 raises questions as to whether the observed effects stem from fullerenes or from the organic solvents used during the preparation of the suspensions. In this paper, we set out to elucidate the mechanism by which tetrahydrofuran (THF) treatment to enhance the preparation of nC60 leads to cytotoxicity in a mouse macrophage cell line. Our results demonstrate that THF/nC60 but not fullerol or aqueous nC60 generates cellular toxicity through a pathway that involves increased intracellular flux and mitochondrial perturbation in RAW 264.7 cells. Interestingly, the supernatant of the THF/n60 suspension rather than the colloidal fullerene aggregates mimics the cytotoxic effects due to the presence of gamma-butyrolactone and formic acid. Thus, the role of nC60 in the cellular responses is likely not due to the direct effect of the nC60 material surface on the cells but is related to the conversion of THF into a toxic byproduct during preparation of the suspension.

Comparative photoactivity and antibacterial properties of C60 fullerenes and titanium dioxide nanoparticles.

The production of reactive oxygen species (ROS) by aqueous suspensions of fullerenes and nano-TiO2 (Degussa P25) was measured both in ultrapure water and in minimal Davis (MD) microbial growth medium. Fullerol (hydroxylated C60) produced singlet oxygen (1O2) in ultrapure water and both 1O2 and superoxide (O2-*) in MD medium, but no hydroxyl radicals (OH*) were detected in either case. PVP/C60 (C60 encapsulated with poly(N-vinylpyrrolidone)) was more efficient than fullerol in generating singlet oxygen and superoxide. However, two other aggregates of C60, namely THF/nC60 (prepared with tetrahydofuran as transitional solvent) and aqu/nC60 (prepared by vigorous stirring of C60 powder in water), were not photoactive. Nano-TiO2 (also present as aggregates) primarily produced hydroxyl radicals in pure water and superoxide in MD medium. Bacterial (Escherichia coli) toxicity tests suggest that, unlike nano-TiO2 which was exclusively phototoxic, the antibacterial activity of fullerene suspensions was linked to ROS production. Nano-TiO2 may be more efficient for water treatment involving UV or solar energy, to enhance contaminant oxidation and perhaps for disinfection. However, fullerol and PVP/ C60 may be useful as water treatment agents targeting specific pollutants or microorganisms that are more sensitive to either superoxide or singlet oxygen.

Mechanisms of photochemistry and reactive oxygen production by fullerene suspensions in water.

Buckminsterfullerene (C60) is a known photosensitizer that produces reactive oxygen species (ROS) in the presence of light; however, its properties in aqueous environments are still not well understood or modeled. In this study, production of both singlet oxygen and superoxide by UV photosensitization of colloidal aggregates of C60 in water was measured by two distinct methods: electron paramagnetic resonance (EPR) with a spin trapping compound, and spectrophotometric detection of the reduced form of the tetrazolium compound XTT. Both singlet oxygen and superoxide were generated by fullerol suspensions while neither was detected in the aqu/nC60 suspensions. A mechanistic framework for photosensitization that takes into account differences in C60 aggregate structure in water is proposed to explain these results. While theory developed for single molecules suggests that alterations to the C60 cage should reduce the quantum yield for the triplet state and associated ROS production, the failure to detect ROS production by aqu/nC60 is explained in part by a more dense aggregate structure compared with the hydroxylated C60.

Transport and retention of colloidal aggregates of C60 in porous media: effects of organic macromolecules, ionic composition, and preparation method.

The physical-chemical behavior of the fullerene C60 in environmental and physiological media is of interest for understanding the potential transport, exposure, and impacts of these materials on organisms and ecosystems. We considerthe role of electrolyte composition and concentration, the effect of organic macromolecules, and the mode of preparation of colloidal aggregates of C60 (nC60) on the deposition of these colloids in a porous medium such as a groundwater aquifer or a water treatment filter. Results for nC60 deposition are qualitatively consistent with trends anticipated by theory. Deposition was found to increase with increasing ionic strength, the presence of polysaccharide-type organic matter, and lower Darcy velocities. Factors that will tend to decrease the retention of these materials in porous media include a low ionic strength and the presence of humic-like substances, while the ionic strengths typical of many natural waters and the presence polysaccharide-based natural organic matter, as may be produced by algae or bacteria, will tend to favor deposition and reduced potential for exposure. Variability in the method of preparing colloidal aggregates of fullerenes was observed to yield significant differences in nC60 properties and transport behavior.

Inactivation of bacteriophages via photosensitization of fullerol nanoparticles.

The production of two reactive oxygen species through UV photosensitization of polyhydroxylated fullerene (fullerol) is shown to enhance viral inactivation rates. The production of both singlet oxygen and superoxide by fullerol in the presence of UV light is confirmed via two unique methods: electron paramagnetic resonance and reduction of nitro blue tetrazolium. These findings build on previous results both in the area of fullerene photosensitization and in the area of fullerene impact on microfauna. Results showed thatthe first-order MS2 bacteriophage inactivation rate nearly doubled due to the presence of singlet oxygen and increased by 125% due to singlet oxygen and superoxide as compared to UVA illumination alone. When fullerol and NADH are present in solution, dark inactivation of viruses occurs at nearly the same rate as that produced by UVA illumination without nanoparticles. These results suggest a potential for fullerenes to impact virus populations in both natural and engineered systems ranging from surface waters to disinfection technologies for water and wastewater treatment.