Smallest unit of maximal entropy as novel experimental criterion for parametric characterization of middle- and high-entropy materials.

Materials with multiple principal elements (middle- and high-entropy materials), are used in emerging applications in various fields due to their unique properties, driven by configuration entropy. Improved understanding and experimental investigations of the impact of the entropy of mixing on the properties of these materials are of large practical interest. Here we show a simplified limited area calculation approach for assessing the entropy of mixing using a CoCuFeNi model nanoalloy. Based on our calculations we propose a new parametric entropy-based criterion, which defines critical scale parameter transition from the maximal entropy state to the entropy-depleted state of the system. The criterion could be used for generalized mechanistic assessment of the effect of the entropy of mixing on the characteristics of the materials with multiple principal elements and for the development and characterization of existing and new middle- and high-entropy materials with both simple single-, and more complex, multiple-sublattice structures.

Eco-corona-mediated transformation of nano-sized Y2O3 in simulated freshwater: A short-term study.

The use of metal and metal oxide nanomaterials (NMs) is experiencing a significant surge in popularity due to their distinctive structures and properties, making them highly attractive for a wide range of applications. This increases the risks of their potential negative impact on organisms if dispersed into the environment. Information about their behavior and transformation upon environmental interactions in aquatic settings is limited. In this study, the influence of naturally excreted biomolecules from the zooplankton Daphnia magna on nanosized Y2O3 of different concentrations was systematically examined in synthetic freshwater in terms of adsorption and eco-corona formation, colloidal stability, transformation, dissolution, and ecotoxicity towards D. magna. The formation of an eco-corona on the surface of the Y2O3 NMs leads to improved colloidal stability and a reduced extent of dissolution. Exposure to the Y2O3 NMs lowered the survival probability of D. magna considerably. The ecotoxic potency was slightly reduced by the formation of the eco-corona, though shown to be particle concentration-specific. Overall, the results highlight the importance of systematic mechanistic and fundamental studies of factors that can affect the environmental fate and ecotoxic potency of NMs.

Unravelling the Mechanistic Understanding of Metal Nanoparticle-Induced Reactive Oxygen Species Formation: Insights from a Cu Nanoparticle Study.

Humans can be exposed to engineered and nonintentionally formed metal and metal oxide nanoparticles (Me NPs) in occupational settings, in public transportation areas, or by means of contact with different consumer products. A critical factor in the toxic potency of Me NPs is their ability to induce oxidative stress. It is thus essential to assess the potential reactive oxygen species (ROS) formation properties of Me NPs. A common way to assess the relative extent of ROS formation in vitro is to use fluorescence spectroscopy with the DCFH-DA (2',7'-dichlorofluorescein diacetate) probe, with and without HRP (horseradish peroxidase). However, this method does not provide any information about specific ROS species or reaction mechanisms. This study investigated the possibility of using complementary techniques to obtain more specific information about formed ROS species, both the type and reaction mechanisms. Cu NPs in PBS (phosphate buffered saline) were chosen as a test system to have the simplest (least interference from other components) aqueous solution with a physiologically relevant pH. ROS formation was assessed using fluorescence by means of the DCFH-DA method (information on relative amounts of oxygen radicals without selectivity), the Ghormley's triiodide method using UV-vis spectrophotometry (concentrations of H2O2), and electron paramagnetic resonance with DMPO as the spin-trap agent (information on specific oxygen radicals). This approach elucidates that Cu NPs undergo ROS-generating corrosion reactions, which previously have not been assessed in situ. In the presence of H2O2, and based on the type of oxygen radical formed, it was concluded that released copper participates in Haber-Weiss and/or Fenton reactions rather than in Fenton-like reactions. The new combination of techniques used to determine ROS induced by Me NPs provides a way forward to gain a mechanistic understanding of Me NP-induced ROS formation, which is important for gaining crucial insight into their ability to induce oxidative stress.

Health hazards of particles in additive manufacturing: a cross-disciplinary study on reactivity, toxicity and occupational exposure to two nickel-based alloys.

The increasing use of additive manufacturing (AM) techniques (e.g., 3D-printing) offers many advantages but at the same time presents some challenges. One concern is the possible exposure and health risk related to metal containing particles of different sizes. Using the nickel-based alloys Hastelloy X (HX) and Inconel 939 (IN939) as a case, the aim of this cross-disciplinary study was to increase the understanding on possible health hazards and exposure. This was done by performing in-depth characterization of virgin, reused and condensate powders, testing in vitro toxicity (cytotoxicity, genotoxicity, oxidative stress), and measuring occupational airborne exposure. The results showed limited metal release from both HX and IN939, and slightly different surface composition of reused compared to virgin powders. No or small effects on the cultured lung cells were observed when tested up to 100 µg/mL. Particle background levels in the printing facilities were generally low, but high transient peaks were observed in relation to sieving. Furthermore, during post processing with grinding, high levels of nanoparticles (> 100,000 particles/cm3) were noted. Urine metal levels in AM operators did not exceed biomonitoring action limits. Future studies should focus on understanding the toxicity of the nanoparticles formed during printing and post-processing.

Characterization and Toxic Potency of Airborne Particles Formed upon Waste from Electrical and Electronic Equipment Waste Recycling: A Case Study.

Manual dismantling, shredding, and mechanical grinding of waste from electrical and electronic equipment (WEEE) at recycling facilities inevitably lead to the accidental formation and release of both coarse and fine particle aerosols, primarily into the ambient air. Since diffuse emissions to air of such WEEE particles are not regulated, their dispersion from the recycling plants into the adjacent environment is possible. The aim of this interdisciplinary project was to collect and characterize airborne WEEE particles smaller than 1 µm generated at a Nordic open waste recycling facility from a particle concentration, shape, and bulk and surface composition perspective. Since dispersed airborne particles eventually may reach rivers, lakes, and possibly oceans, the aim was also to assess whether such particles may pose any adverse effects on aquatic organisms. The results show that WEEE particles only exerted a weak tendency toward cytotoxic effects on fish gill cell lines, although the exposure resulted in ROS formation that may induce adverse effects. On the contrary, the WEEE particles were toxic toward the crustacean zooplankter Daphnia magna, showing strong effects on survival of the animals in a concentration-dependent way.

The effect of natural biomolecules on yttrium oxide nanoparticles from a Daphnia magna survival rate perspective.

The attention to rare earth oxide nanoparticles (NPs), including yttrium oxide (Y2O3), has increased in many fields due to their unique structural characteristics and functional properties. The aim of our study was to investigate the mechanisms by which bio-corona formation on Y2O3 NPs affects their environmental fate and toxicity. The Y2O3 NPs induced toxicity to freshwater filter feeder Daphnia magna at particle concentrations of 1 and 10 mg/L, regardless of particle size. Interactions between naturally excreted biomolecules (e.g. protein, lipids, and polysaccharides) derived from D. magna, and the Y2O3 NPs (30-45 nm) resulted in the formation of an eco-corona, which reduced their toxic effects toward D. magna at a particle concentration of 10 mg/L. No effects were observed at lower concentrations or for the other particle sizes investigated. Copper-zinc (Cu-Zn) superoxide dismutase, apolipophorins, and vitellogenin-1 proteins proved to be the most prominent proteins of the adsorbed corona, and possibly a reason for the reduced toxicity of the 30-45 nm Y2O3 NPs toward D. magna.

Effects of interactions between natural organic matter and aquatic organism degradation products on the transformation and dissolution of cobalt and nickel-based nanoparticles in synthetic freshwater.

Expanding applications and production of engineered nanoparticles lead to an increased risk for their environmental dispersion. Systematic knowledge of surface transformation and dissolution of nanoparticles is essential for risk assessment and regulation establishment. Such aspects of Co- and Ni-based nanoparticles including metals, oxides, and solution combustion synthesized metal nanoparticles (metal cores with carbon shells) were investigated upon environmental interaction with organic matter, simulated by natural organic matter (NOM) and degradation products from zooplankton and algae (eco-corona biomolecules, EC) in freshwater (FW). The presence of NOM and EC in FW results in negative surface charges of the nanoparticles reduces the extent of nanoparticles agglomeration, and increases concentration, mainly due to the surface adsorption of carboxylate groups of the organic matter. The dissolution of the Co-based nanoparticles was for all conditions (FW, FW with NOM or EC) higher than the Ni-based, except for Co3O4 being nearly non-soluble. The surface transformation and dissolution of nanoparticles are highly exposure and time-dependent, and surface- and environment specific. Therefore, no general correlation was observed between dissolution and, particle types, surface conditions, or EC/NOM adsorption. This underlines the importance of thorough investigations of nanoparticles adsorption/desorption, degradation, and exposure scenarios for developing regulatory relevant protocols and guidelines.

A Novel Nanosafety Approach Using Cell Painting, Metabolomics, and Lipidomics Captures the Cellular and Molecular Phenotypes Induced by the Unintentionally Formed Metal-Based (Nano)Particles.

Additive manufacturing (AM) or industrial 3D printing uses cutting-edge technologies and materials to produce a variety of complex products. However, the effects of the unintentionally emitted AM (nano)particles (AMPs) on human cells following inhalation, require further investigations. The physicochemical characterization of the AMPs, extracted from the filter of a Laser Powder Bed Fusion (L-PBF) 3D printer of iron-based materials, disclosed their complexity, in terms of size, shape, and chemistry. Cell Painting, a high-content screening (HCS) assay, was used to detect the subtle morphological changes elicited by the AMPs at the single cell resolution. The profiling of the cell morphological phenotypes, disclosed prominent concentration-dependent effects on the cytoskeleton, mitochondria, and the membranous structures of the cell. Furthermore, lipidomics confirmed that the AMPs induced the extensive membrane remodeling in the lung epithelial and macrophage co-culture cell model. To further elucidate the biological mechanisms of action, the targeted metabolomics unveiled several inflammation-related metabolites regulating the cell response to the AMP exposure. Overall, the AMP exposure led to the internalization, oxidative stress, cytoskeleton disruption, mitochondrial activation, membrane remodeling, and metabolic reprogramming of the lung epithelial cells and macrophages. We propose the approach of integrating Cell Painting with metabolomics and lipidomics, as an advanced nanosafety methodology, increasing the ability to capture the cellular and molecular phenotypes and the relevant biological mechanisms to the (nano)particle exposure.

Reactive Oxygen Species Formed by Metal and Metal Oxide Nanoparticles in Physiological Media-A Review of Reactions of Importance to Nanotoxicity and Proposal for Categorization.

Diffusely dispersed metal and metal oxide nanoparticles (NPs) can adversely affect living organisms through various mechanisms and exposure routes. One mechanism behind their toxic potency is their ability to generate reactive oxygen species (ROS) directly or indirectly to an extent that depends on the dose, metal speciation, and exposure route. This review provides an overview of the mechanisms of ROS formation associated with metal and metal oxide NPs and proposes a possible way forward for their future categorization. Metal and metal oxide NPs can form ROS via processes related to corrosion, photochemistry, and surface defects, as well as via Fenton, Fenton-like, and Haber-Weiss reactions. Regular ligands such as biomolecules can interact with metallic NP surfaces and influence their properties and thus their capabilities of generating ROS by changing characteristics such as surface charge, surface composition, dissolution behavior, and colloidal stability. Interactions between metallic NPs and cells and their organelles can indirectly induce ROS formation via different biological responses. H2O2 can also be generated by a cell due to inflammation, induced by interactions with metallic NPs or released metal species that can initiate Fenton(-like) and Haber-Weiss reactions forming various radicals. This review discusses these different pathways and, in addition, nano-specific aspects such as shifts in the band gaps of metal oxides and how these shifts at biologically relevant energies (similar to activation energies of biological reactions) can be linked to ROS production and indicate which radical species forms. The influences of kinetic aspects, interactions with biomolecules, solution chemistry (e.g., Cl- and pH), and NP characteristics (e.g., size and surface defects) on ROS mechanisms and formation are discussed. Categorization via four tiers is suggested as a way forward to group metal and metal oxide NPs based on the ROS reaction pathways that they may undergo, an approach that does not include kinetics or environmental variations. The criteria for the four tiers are based on the ability of the metallic NPs to induce Fenton(-like) and Haber-Weiss reactions, corrode, and interact with biomolecules and their surface catalytic properties. The importance of considering kinetic data to improve the proposed categorization is highlighted.

Influence of natural organic matter on the transformation of metal and metal oxide nanoparticles and their ecotoxic potency in vitro.

Increased use and production of engineered nanoparticles (NPs) lead to an elevated risk of their diffuse dispersion into the aquatic environment and increased concern on unknown effects induced by their release into the aquatic ecosystem. An improved understanding of the environmental transformation processes of NPs of various surface characteristics is hence imperative for risk assessment and management. This study presents results on effects of natural organic matter (NOM) on the environmental transformation and dissolution of metal and metal oxide NPs of different surface and solubility properties in synthetic freshwater (FW) with and without NOM. Adsorption of NOM was evident on most of the studied NPs, except Sb and Sb2O3, which resulted in the formation of negatively charged colloids of higher stability and smaller size distribution compared with the same NPs in FW only. The dissolution rate of the NPs in the presence of NOM correlated with the strength of interactions between the carboxylate group of NOM and the particle surface, and resulted in either no (Mn, Sb, ZnO NPs), increased (Co, Sn NPs) and decreased (Ni, NiO, Sb2O3, Y2O3 NPs) levels of dissolution. One type of metal NP from each group (Mn, Ni, Sn) were investigated to assess whether observed differences in adsorption of NOM and dissolution would influence their ecotoxic potency. The results showed Mn, Ni, and Sn NPs to generate intracellular reactive oxygen species (ROS) in a time and dose-dependent manner. The extent of ROS generation in FW was similar for both Mn and Ni NPs but higher for Sn NPs. These findings are possibly related to interactions and infiltration of the NPs with the cells, which lead to redox imbalances which could induce oxidative stress and cell damage. At the same time, the presence of NOM generally reduced the intracellular ROS generation by 20-40% for the investigated NPs and also reduced cytotoxicity of Sn NPs, which can be attributed to the stronger interaction of carboxylate groups of NOM with the surface of the NPs.

SLM-processed MoS2/Mo2S3 nanocomposite for energy conversion/storage applications.

MoS2-based nanocomposites have been widely processed by a variety of conventional and 3D printing techniques. In this study, selective laser melting (SLM) has for the first time successfully been employed to tune the crystallographic structure of bulk MoS2 to a 2H/1T phase and to distribute Mo2S3 nanoparticles in-situ in MoS2/Mo2S3 nanocomposites used in electrochemical energy conversion/storage systems (EECSS). The remarkable results promote further research on and elucidate the applicability of laser-based powder bed processing of 2D nanomaterials for a wide range of functional structures within, e.g., EECSS, aerospace, and possibly high-temperature solid-state EECSS even in space.

Toxicity evaluation of particles formed during 3D-printing: Cytotoxic, genotoxic, and inflammatory response in lung and macrophage models.

Additive manufacturing (AM) or "3D-printing" is a ground-breaking technology that enables the production of complex 3D parts. Its rapid growth calls for immediate toxicological investigations of possible human exposures in order to estimate occupational health risks. Several laser-based powder bed fusion AM techniques are available of which many use metal powder in the micrometer range as feedstock. Large energy input from the laser on metal powders generates several by-products, like spatter and condensate particles. Due to often altered physicochemical properties and composition, spatter and condensate particles can result in different toxicological responses compared to the original powder particles. The toxicity of such particles has, however, not yet been investigated. The aim of the present study was to investigate the toxicity of condensate/spatter particles formed and collected upon selective laser melting (SLM) printing of metal alloy powders, including a nickel-chromium-based superalloy (IN939), a nickel-based alloy (Hastelloy X, HX), a high-strength maraging steel (18Ni300), a stainless steel (316L), and a titanium alloy (Ti6Al4V). Toxicological endpoints investigated included cytotoxicity, generation of reactive oxygen species (ROS), genotoxicity (comet and micronucleus formation), and inflammatory response (cytokine/chemokine profiling) following exposure of human bronchial epithelial cells (HBEC) or monocytes/macrophages (THP-1). The results showed no or minor cytotoxicity in the doses tested (10-100 µg/mL). Furthermore, no ROS generation or formation of micronucleus was observed in the HBEC cells. However, an increase in DNA strand breaks (detected by comet assay) was noted in cells exposed to HX, IN939, and Ti6Al4V, whereas no evident release of pro-inflammatory cytokine was observed from macrophages. Particle and surface characterization showed agglomeration in solution and different surface oxide compositions compared to the nominal bulk content. The extent of released nickel was small and related to the nickel content of the surface oxides, which was largely different from the bulk content. This may explain the limited toxicity found despite the high Ni bulk content of several powders. Taken together, this study suggests relatively low acute toxicity of condensates/spatter particles formed during SLM-printing using IN939, HX, 18Ni300, 316L, and Ti6Al4V as original metal powders.

High-Resolution Microscopical Studies of Contact Killing Mechanisms on Copper-Based Surfaces.

The mechanisms of bacterial contact killing induced by Cu surfaces were explored through high-resolution studies based on combinations of the focused ion beam (FIB), scanning transmission electron microscopy (STEM), high-resolution TEM, and nanoscale Fourier transform infrared spectroscopy (nano-FTIR) microscopy of individual bacterial cells of Gram-positive Bacillus subtilis in direct contact with Cu metal and Cu5Zn5Al1Sn surfaces after high-touch corrosion conditions. This approach permitted subcellular information to be extracted from the bioinorganic interface between a single bacterium and Cu/Cu5Zn5Al1Sn surfaces after complete contact killing. Early stages of interaction between individual bacteria and the metal/alloy surfaces include cell leakage of extracellular polymeric substances (EPSs) from the bacterium and changes in the metal/alloy surface composition upon adherence of bacteria. Three key observations responsible for Cu-induced contact killing include cell membrane damage, formation of nanosized copper-containing particles within the bacteria cell, and intracellular copper redox reactions. Direct evidence of cell membrane damage was observed upon contact with both Cu metal and Cu5Zn5Al1Sn surfaces. Cell membrane damage permits copper to enter into the cell interior through two possible routes, as small fragmentized Cu2O particles from the corrosion product layer and/or as released copper ions. This results in the presence of intracellular copper oxide nanoparticles inside the cell. The nanosized particles consist primarily of CuO with smaller amounts of Cu2O. The existence of two oxidation states of copper suggests that intracellular redox reactions play an important role. The nanoparticle formation can be regarded as a detoxification process of copper, which immobilizes copper ions via transformation processes within the bacteria into poorly soluble or even insoluble nanosized Cu structures. Similarly, the formation of primarily Cu(II) oxide nanoparticles could be a possible way for the bacteria to deactivate the toxic effects induced by copper ions via conversion of Cu(I) to Cu(II).

Genotoxicity and inflammatory potential of stainless steel welding fume particles: an in vitro study on standard vs Cr(VI)-reduced flux-cored wires and the role of released metals.

Welders are daily exposed to various levels of welding fumes containing several metals. This exposure can lead to an increased risk for different health effects which serves as a driving force to develop new methods that generate less toxic fumes. The aim of this study was to explore the role of released metals for welding particle-induced toxicity and to test the hypothesis that a reduction of Cr(VI) in welding fumes results in less toxicity by comparing the welding fume particles of optimized Cr(VI)-reduced flux-cored wires (FCWs) to standard FCWs. The welding particles were thoroughly characterized, and toxicity (cell viability, DNA damage and inflammation) was assessed following exposure to welding particles as well as their released metal fraction using cultured human bronchial epithelial cells (HBEC-3kt, 5-100 µg/mL) and human monocyte-derived macrophages (THP-1, 10-50 µg/mL). The results showed that all Cr was released as Cr(VI) for welding particles generated using standard FCWs whereas only minor levels (< 3% of total Cr) were released from the newly developed FCWs. Furthermore, the new FCWs were considerably less cytotoxic and did not cause any DNA damage in the doses tested. For the standard FCWs, the Cr(VI) released in cell media seemed to explain a large part of the cytotoxicity and DNA damage. In contrast, all particles caused rather similar inflammatory effects suggesting different underlying mechanisms. Taken together, this study suggests a potential benefit of substituting standard FCWs with Cr(VI)-reduced wires to achieve less toxic welding fumes and thus reduced risks for welders.

Adsorption of Horseradish Peroxidase on Metallic Nanoparticles: Effects on Reactive Oxygen Species Detection Using 2',7'-Dichlorofluorescin Diacetate.

The fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) together with the enzyme horseradish peroxidase (HRP) is widely used in nanotoxicology to study acellular reactive oxygen species (ROS) production from nanoparticles (NPs). This study examined whether HRP adsorbs onto NPs of Mn, Ni, and Cu and if this surface process influences the extent of metal release and hence the ROS production measurements using the DCFH assay in phosphate buffered saline (PBS), saline, or Dulbecco's modified Eagle's medium (DMEM). Adsorption of HRP was evident onto all NPs and conditions, except for Mn NPs in PBS. The presence of HRP resulted in an increased release of copper from the Cu NPs in PBS and reduced levels of nickel from the Ni NPs in saline. Both metal ions in solution and the adsorption of HRP onto the NPs can change the activity of HRP and thus influence the ROS results. The effect of HRP on the NP reactivity was shown to be solution chemistry dependent. Most notable was the evident affinity/adsorption of phosphate toward the metal NPs, followed by a reduced adsorption of HRP, the concomitant reduction in released manganese from the Mn NPs, and increased levels of released metals from the Cu NPs in PBS. Minor effects were observed for the Ni NPs. The solution pH should be monitored since the release of metals can change the solution pH and the activity of HRP is known to be pH-dependent. It is furthermore essential that solution pH adjustments are made following the addition of NaOH during diacetyl removal of DCFH-DA. Even though not observed for the given exposure conditions of this study, released metal ions could possibly induce agglomeration or partial denaturation of HRP, which in turn could result in steric hindrance for H2O2 to reach the active site of HRP. This study further emphasizes the influence of HRP on the background kinetics, its solution dependence, and effects on measured ROS signals. Different ways of correcting for the background are highlighted, as this can result in different interpretations of generated results. The results show that adsorption of HRP onto the metal NPs influenced the extent of metal release and may, depending on the investigated system, result in either under- or overestimated ROS signals if used together with the DCFH assay. HRP should hence be used with caution when measuring ROS in the presence of reactive metallic NPs.

Corrosion and transformation of solution combustion synthesized Co, Ni and CoNi nanoparticles in synthetic freshwater with and without natural organic matter.

Pure metallic Co, Ni, and their bimetallic compositions of Co3Ni, CoNi, and CoNi3 nanomaterials were prepared by solution combustion synthesis. Microstructure, phase composition, and crystalline structure of these nanoparticles (NPs) were characterized along with studies of their corrosion and dissolution properties in synthetic freshwater with and without natural organic matter (NOM). The nanomaterials consisted of aggregates of fine NPs (3-30 nm) of almost pure metallic and bimetallic crystal phases with a thin surface oxide covered by a thin carbon shell. The nanomaterials were characterized by BET surface areas ranging from ~ 1 to 8 m2/g for the Ni and Co NPs, to 22.93 m2/g, 14.86 m2/g, and 10.53 m2/g for the Co3Ni, CoNi, CoNi3 NPs, respectively. More Co and Ni were released from the bimetallic NPs compared with the pure metals although their corrosion current densities were lower. In contrast to findings for the pure metal NPs, the presence of NOM increased the release of Co and Ni from the bimetallic NPs in freshwater compared to freshwater only even though its presence reduced the corrosion rate (current density). It was shown that the properties of the bimetallic nanomaterials were influenced by multiple factors such as their composition, including carbon shell, type of surface oxides, and the entropy of mixing.