Behavior Of Engineered Nanomaterials Research Articles

Novel multimethod approach for the determination of the colloidal stability of nanomaterials in complex environmental mixtures using a global stability index: TiO2 as case study

A systematic study on the colloidal behavior of uncoated and polyvinylpyrrolidone (PVP) coated TiO2 engineered nanomaterials (ENMs) in simulated aqueous media is herein reported, in which conditions representative for natural waters (pH, presence of divalent electrolytes (i.e. Ca2+/Mg2+ and SO42−), of natural organic matter (NOM) and of suspended particulate matter (SPM)) were systematically varied. The colloidal stability of the different dispersions was investigated by means of Dynamic and Electrophoretic Light Scattering (DLS and ELS) and Centrifugal Separation Analysis (CSA), and a global stability index based on these three techniques was developed. The index allows to quantitatively classify the nano-based dispersions according to their colloidal stability affected by the different parameters studied. This multimethod approach clearly identifies inorganic SPM followed by divalent electrolytes as the main natural components destabilizing TiO2 ENMs upon entering in simulated natural waters, while it highlights a moderate stabilization induced by NOM, depending mainly on pH. Moreover, the PVP coating was found to attenuate the influence of these parameters on the colloidal stability. The obtained results show how the global stability index developed is influenced by the complexity of the system, suggesting the importance of combining the information gathered from all the techniques employed to better elucidate the fate and behavior of ENMs in natural surface waters.

The impacts of metal-based engineered nanomaterial mixtures on microbial systems: A review.

The last decade has witnessed tremendous growth in the commercial use of metal-based engineered nanomaterials (ENMs) for a wide range of products and processes. Consequently, direct and indirect release into environmental systems may no longer be considered negligible or insignificant. Yet, there is an active debate as to whether there are real risks to human or ecological health with environmental exposure to ENMs. Previous research has focused primarily on the acute effects of individual ENMs using pure cultures under controlled laboratory environments, which may not accurately reveal the ecological impacts of ENMs under real environmental conditions. The goal of this review is to assess our current understanding of ENM effects as we move from exposure of single to multiple ENMs or microbial species. For instance, are ENMs' impacts on microbial communities predicted by their intrinsic physical or chemical characteristics or their effects on single microbial populations; how do chronic ENM interactions compare to acute toxicity; does behavior under simplified laboratory conditions reflect that in environmental media; finally, is biological stress modified by interactions in ENM mixtures relative to that of individual ENM? This review summarizes key findings and our evolving understanding of the ecological effects of ENMs under complex environmental conditions on microbial systems, identifies the gaps in our current knowledge, and indicates the direction of future research.

MESOCOSM: A mesocosm database management system for environmental nanosafety

Engineered nanomaterials (ENMs) are intentionally designed and produced by humans to revolutionize the manufacturing sector, such as electronic goods, paints, tires, clothes, cosmetic products, and biomedicine. With the spread of these ENMs in our daily lives, scientific research have generated a huge amount of data related to their potential impacts on human and environment health. To date, these data are gathered in databases mainly focused on the (eco)toxicity and occupational exposure to ENMs. These databases are therefore not suitable to build well-informed environmental exposure scenarios covering the life cycle of ENMs. In this paper, we report the construction of one of the first centralized mesocosm database management system for environmental nanosafety (called MESOCOSM) containing experimental data collected from mesocosm experiments suited for understanding and quantifying both the environmental hazard and exposure. The database, which is publicly available through https://aliayadi.github.io/MESOCOSM-database/, contains 5200 entities covering tens of unique experiments investigating Ag, CeO2, CuO, TiO2-based ENMs as well as nano-enabled products. These entities are divided into different groups i.e. physicochemical properties of ENMS, environmental, exposure and hazard endpoints, and other general information about the mesocosm testing, resulting in more than forty parameters in the database. The MESOCOSM database is equipped with a powerful application, consisting of a graphical user interface (GUI), allowing users to manage and search data using complex queries without relying on programmers. MESOCOSM aims to predict and explain ENMs behavior and fate in different ecosystems as well as their potential impacts on the environment at different stages of the nanoproducts lifecycle. MESOCOSM is expected to benefit the nanosafety community by providing a continuous source of critical information and additional characterization factors for predicting ENMs interactions with the environment and their risks.

The Crucial Role of Environmental Coronas in Determining the Biological Effects of Engineered Nanomaterials.

In aquatic environments, a large number of ecological macromolecules (e.g., natural organic matter (NOM), extracellular polymeric substances (EPS), and proteins) can adsorb onto the surface of engineered nanomaterials (ENMs) to form a unique environmental corona. The presence of environmental corona as an eco-nano interface can significantly alter the bioavailability, biocompatibility, and toxicity of pristine ENMs to aquatic organisms. However, as an emerging field, research on the impact of the environmental corona on the fate and behavior of ENMs in aquatic environments is still in its infancy. To promote a deeper understanding of its importance in driving or moderating ENM toxicity, this study systemically recapitulates the literature of representative types of macromolecules that are adsorbed onto ENMs; these constitute the environmental corona, including NOM, EPS, proteins, and surfactants. Next, the ecotoxicological effects of environmental corona-coated ENMs on representative aquatic organisms at different trophic levels are discussed in comparison to pristine ENMs, based on the reported studies. According to this analysis, molecular mechanisms triggered by pristine and environmental corona-coated ENMs are compared, including membrane adhesion, membrane damage, cellular internalization, oxidative stress, immunotoxicity, genotoxicity, and reproductive toxicity. Finally, current knowledge gaps and challenges in this field are discussed from the ecotoxicology perspective.

Nanotechnology in soil remediation - applications vs. implications.

Engineered nanomaterials (ENMs) and nanotechnology have shown great potential in addressing complex problems and creating innovative approaches in soil remediation due to their unique features of high reactivity, selectivity and versatility. Meanwhile, valid concerns exist with regard to their implications towards the terrestrial environment and the ecosystem. This review summarizes: (i) the applications and the corresponding mechanisms of various types of ENMs for soil remediation; (ii) the environmental behavior of ENMs in soils and their interactions with the soil content; (iii) the environmental implications of ENMs during remedial applications. The overall objective is to promote responsible innovations so as to take optimal advantage of ENMs and nanotechnology while minimizing their adverse effects to the ecological system. It is critical to establish sustainable remediation methods that ensure a healthy and safe environment without bringing additional risk.

Nanotoxicity assessment: A challenging application for cutting edge electroanalytical tools

In the recent years, the number of commercial products containing engineered nanomaterials (ENMs) has increased exponentially. Consequently, the toxicological profile of ENMs on the ecosystems as well as on human health has to be carefully evaluated. Nanotoxicology, an interdisciplinary research area devoted to assessing the hazards associated with ENMs, is expanding rapidly. Many physicochemical techniques and biochemical methodologies have been proposed and are currently used to characterize nanomaterials from a toxicological point of view. Electroanalytical and bioelectrochemical methods can be useful in expanding the repertoire of accessible nanotoxicity-assessment technologies and to accelerate the testing and screening of the toxicological effects of ENMs. These methods can be used for elucidating the toxicological behavior of ENMs at single cell, cell population and whole-organism levels, for in vitro and in vivo measurements, respectively. The aim of this review is to provide an overview on the bioelectrochemical approaches that have been proposed for ENMs toxicity assessment. Furthermore, an overview on cutting-edge electroanalytical devices with a potential impact to this peculiar application is provided.

Multivariate modeling of engineered nanomaterial features associated with developmental toxicity

Despite the increasing prevalence of engineered nanomaterials (ENMs) in consumer products, their toxicity profiles remain to be elucidated. ENM physicochemical characteristics (PCC) are known to influence ENM behavior, however the mechanisms of these effects have not been quantified. Further confounding the question of how the PCC influence behavior is the inclusion of structural and molecular descriptors in modeling schema that minimize the effects of PCC on the toxicological endpoints. In this work, we analyze ENM physico-chemical measurements that have not previously been studied within a developmental toxicity framework using an embryonic zebrafish model. In testing a panel of diverse ENMs to build a consensus model, we found nonlinear relationships between any singular PCC and bioactivity. By using a machine learning (ML) method to characterize the information content of combinatorial PCC sets, we found that concentration, surface area, shape, and polydispersity can accurately capture the developmental toxicity profile of ENMs with consideration to whole-organism effects.

INSIdE NANO: a systems biology framework to contextualize the mechanism-of-action of engineered nanomaterials

Engineered nanomaterials (ENMs) are widely present in our daily lives. Despite the efforts to characterize their mechanism of action in multiple species, their possible implications in human pathologies are still not fully understood. Here we performed an integrated analysis of the effects of ENMs on human health by contextualizing their transcriptional mechanism-of-action with respect to drugs, chemicals and diseases. We built a network of interactions of over 3,000 biological entities and developed a novel computational tool, INSIdE NANO, to infer new knowledge about ENM behavior. We highlight striking association of metal and metal-oxide nanoparticles and major neurodegenerative disorders. Our novel strategy opens possibilities to achieve fast and accurate read-across evaluation of ENMs and other chemicals based on their biosignatures.

Comparative differences in the behavior of TiO2 and SiO2 food additives in food ingredient solutions

Nanotechnology is widely used in the food industry to improve the color, taste, and texture of food products. However, concerns regarding potential undesirable health effects remain. It is expected that interaction of engineered nanomaterials (ENMs) with food ingredients will influence their behavior and the resulting corona. Nonetheless, there are limited systematic studies conducted to clarify this understanding to date. Herein, we investigated the behavior and corona formation of food grade titanium dioxide (TiO2) and silicon dioxide (SiO2) in solutions of model food ingredients including bovine serum albumin (BSA) and sucrose. Measurements using dynamic light scattering (DLS) showed that both TiO2 and SiO2 nanoparticles displayed a decrease in agglomerate sizes in the presence of both food ingredients. Both particles were negatively charged in all the conditions tested. Corona adsorption studies were carried out using multiple complementary methods including Fourier transform infrared (FTIR) spectroscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS), transmission electron microscopy (TEM), micro bicinchoninic acid (BCA) protein assay, and thermogravimetric analysis (TGA). Comparative investigation showed that sucrose could disperse both particles more effectively than BSA and that SiO2 displayed greater adsorption capacity for both BSA and sucrose, compared to TiO2. Taken collectively, this study demonstrated the importance of considering food ingredient effects when mapping the behavior of ENMs in food products. Such understanding could be significant in the evaluation of biological effects, such as toxicity, of ENMs used in food products.

The impact of nanomaterial characteristics on inhalation toxicity.

During the last few decades, nanotechnology has evolved into a success story, apparent from a steadily increasing number of scientific publications as well as a large number of applications based on engineered nanomaterials (ENMs). Its widespread uses suggest a high relevance for consumers, workers and the environment, hence justifying intensive investigations into ENM-related adverse effects as a prerequisite for nano-specific regulations. In particular, the inhalation of airborne ENMs, being assumed to represent the most hazardous type of human exposure to these kinds of particles, needs to be scrutinized. Due to an increased awareness of possible health effects, which have already been seen in the case of ultrafine particles (UFPs), research and regulatory measures have set in to identify and address toxic implications following their almost ubiquitous occurrence. Although ENM properties differ from those of the respective bulk materials, the available assessment protocols are often designed for the latter. Despite the large benefit ensuing from the application of nanotechnology, many issues related to ENM behavior and adverse effects are not fully understood or should be examined anew. The traditional hypothesis that ENMs exhibit different or additional hazards due to their "nano" size has been challenged in recent years and ENM categorization according to their properties and toxicity mechanisms has been proposed instead. This review summarizes the toxicological effects of inhaled ENMs identified to date, elucidating the modes of action which provoke different mechanisms in the respiratory tract and their resulting effects. By linking particular mechanisms and adverse effects to ENM properties, grouping of ENMs based on toxicity-related properties is supposed to facilitate toxicological risk assessment. As intensive studies are still required to identify these "ENM classes", the need for alternatives to animal studies is evident and advances in cell-based test systems for pulmonary research are presented here. We hope to encourage the ongoing discussion about ENM risks and to advocate the further development and practice of suitable testing and grouping methods.

Importance of exposure dynamics of metal-based nano-ZnO, -Cu and -Pb governing the metabolic potential of soil bacterial communities

Metal-based engineered nanomaterials (ENMs) are known to affect bacterial processes and metabolic activities. While testing their negative effects on biological components, studies traditionally rely on initial exposure concentrations and thereby do not take into consideration the dynamic behavior of ENMs that ultimately determines exposure and toxicity (e.g. ion release). Moreover, functional responses of soil microbial communities to ENMs exposure can be caused by both the particulate forms and the ionic forms, yet their relative contributions remain poorly understood. Therefore, we investigated the dynamic changes of exposure concentrations of three different types of ENMs (nano-ZnO, -Cu and -Pb) and submicron particles (SMPs) in relation to their impact on the capacity of soil bacterial communities to utilize carbon substrates. The different ENMs were chosen to differ in dissolution potential. The dynamic exposures of ENMs were considered using a time weighted average (TWA) approach. The joint toxicity of the particulate forms and the ionic forms of ENMs was evaluated using a response addition model. Our results showed that the effect concentrations of spherical nano-ZnO, -Cu and SMPs, and Pb-based perovskites expressed as TWA were lower than expressed as initial concentrations. Both particulate forms and ionic forms of spherical 18nm, 43nm nano-ZnO and 50nm, 100nm nano-Cu contribute to the overall response at the EC50 levels. The particulate forms for 150nm, 200nm and 900nm ZnO SMPs and rod-shaped 78nm nano-Cu mainly affected the soil microbial metabolic potential, while the Cu ions released from spherical 25nm nano-Cu, 500nm Cu SMPs and Pb ions released from perovskites mainly described the effects to bacterial communities. Our results indicate that the dynamic exposure of ENMs and relative contributions of particles and ions require consideration in order to pursue a naturally realistic assessment of environmental risks of metal-based ENMs.

Assessing nanomaterial exposures in aquatic ecotoxicological testing: Framework and case studies based on dispersion and dissolution

The unique behavior of engineered nanomaterials (ENM) in aqueous media and dynamic changes in particle settling, agglomeration and dissolution rates is a challenge to the consistency, reliability and interpretation of standard aquatic hazard bioassay results. While the toxicological endpoints (e.g., survival, growth, reproduction, etc.) in ecotoxicity bioassays are largely applicable to ENMs, the standard methods as written for dissolved substances are confounded by the dynamic settling, agglomeration and dissolution of particulate ENMs during the bioassay. A testing framework was designed to serve as a starting point to identify approaches for the consistent conduct of aquatic hazard tests that account for the behavior of ENMs in test media and suitable data collection to support representative exposure metrology. The framework was demonstrated by conducting three case studies testing ENMs with functionally distinct characteristics and behaviors. Pretests with a temporal sampling of particle concentration, agglomeration and dissolution were conducted on each ENM in test media. Results indicated that a silver nanoparticle (AgNP) powder was not dispersible, a nano-TiO2 powder was dispersible but unstable, and a polyvinylpyrrolidinone-coated AgNP was relatively stable in test media. Based on these functional results, Ceriodaphnia dubia bioassays were conducted to compare different exposure summary methods (nominal, arithmetic average, geometric average, time-weighted average) for calculating and expressing toxicity endpoints. Results indicated that while arithmetic means were effective for expressing the toxicity of more stable materials, time-weighted averaged concentrations were appropriate for the unstable nano-TiO2.

Interpreting unique colloidal response of TiO2 nanomaterials to controlled sonication for understanding of their assembly configuration in water

It is important to determine the assembly configuration of engineered nanomaterials (ENMs) because assembly configuration influences their fate and transport behavior in the aquatic environment. Aggregated particles are more subject to segregation upon changes of environmental conditions (and vice versa) than agglomerated particles. As a strategic tool for investigating the time-resolved reversible segregating and assembling behavior of ENMs and thus estimating their assembly configuration, a controlled sonication process was proposed. It was hypothesized that the unique colloidal response of ENMs to sonication, with respect to changes in size, might be their intrinsic property associated with assembly configuration. As a model ENM, three different TiO2 particles with unique properties (commercial P-25 and UV 100 and home-made (HM) TiO2) were examined with programmed sonication processes under various environmental settings. When they were dispersed in water, all TiO2 particles tested obviously assembled to form much larger clusters. Size of P-25 decreased immediately upon sonication and did not change under the subsequent quiescence step while sizes of UV 100 and HM gradually decreased and then slowly recovered back to their initial sizes. The trend was generally observed in all conditions tested. The unique colloidal response of TiO2 could be explained by its properties associated with assembly configuration.

Dynamic Probabilistic Modeling of Environmental Emissions of Engineered Nanomaterials.

The need for an environmental risk assessment for engineered nanomaterials (ENM) necessitates the knowledge about their environmental concentrations. Despite significant advances in analytical methods, it is still not possible to measure the concentrations of ENM in natural systems. Material flow and environmental fate models have been used to provide predicted environmental concentrations. However, almost all current models are static and consider neither the rapid development of ENM production nor the fact that many ENM are entering an in-use stock and are released with a lag phase. Here we use dynamic probabilistic material flow modeling to predict the flows of four ENM (nano-TiO2, nano-ZnO, nano-Ag and CNT) to the environment and to quantify their amounts in (temporary) sinks such as the in-use stock and ("final") environmental sinks such as soil and sediment. Caused by the increase in production, the concentrations of all ENM in all compartments are increasing. Nano-TiO2 had far higher concentrations than the other three ENM. Sediment showed in our worst-case scenario concentrations ranging from 6.7 μg/kg (CNT) to about 40 000 μg/kg (nano-TiO2). In most cases the concentrations in waste incineration residues are at the "mg/kg" level. The flows to the environment that we provide will constitute the most accurate and reliable input of masses for environmental fate models which are using process-based descriptions of the fate and behavior of ENM in natural systems and rely on accurate mass input parameters.

Aggregation, dissolution, and transformation of copper nanoparticles in natural waters.

Time-dependent aggregation, sedimentation, dissolution, and transformation of three copper-based engineered nanomaterials (ENMs) of varied properties were measured in eight natural and artificial waters. Nano-Cu and Cu(OH)2 aggregated rapidly to >10(3) nm while the aggregate size of nano-CuO averaged between 250 and 400 nm. Aggregate size for both nano-Cu and nano-CuO showed a positive correlation with ionic strength with a few exceptions. Aggregate size did not correlate well with sedimentation rate, suggesting sedimentation was influenced by other factors. Controlling factors in sedimentation rates varied by particle: Cu(OH)2 particles remained stable in all waters but groundwater, nano-Cu was generally unstable except in waters with high organic content, and nano-CuO was stabilized by the presence of phosphate, which reversed surface charge polarity at concentrations as low as 0.1 mg PO4(3-) L(-1). Dissolution generally correlated with pH, although in saline waters, dissolved copper formed insoluble complexes. Nano-Cu was rapidly oxidized, resulting in dissolution immediately followed by the formation of precipitates. These results suggest factors including phosphate, carbonate, and ENM oxidation state may be key in determining Cu ENM behavior in natural waters.

Review of nanomaterials in dentistry: interactions with the oral microenvironment, clinical applications, hazards, and benefits.

Interest in the use of engineered nanomaterials (ENMs) as either nanomedicines or dental materials/devices in clinical dentistry is growing. This review aims to detail the ultrafine structure, chemical composition, and reactivity of dental tissues in the context of interactions with ENMs, including the saliva, pellicle layer, and oral biofilm; then describes the applications of ENMs in dentistry in context with beneficial clinical outcomes versus potential risks. The flow rate and quality of saliva are likely to influence the behavior of ENMs in the oral cavity, but how the protein corona formed on the ENMs will alter bioavailability, or interact with the structure and proteins of the pellicle layer, as well as microbes in the biofilm, remains unclear. The tooth enamel is a dense crystalline structure that is likely to act as a barrier to ENM penetration, but underlying dentinal tubules are not. Consequently, ENMs may be used to strengthen dentine or regenerate pulp tissue. ENMs have dental applications as antibacterials for infection control, as nanofillers to improve the mechanical and bioactive properties of restoration materials, and as novel coatings on dental implants. Dentifrices and some related personal care products are already available for oral health applications. Overall, the clinical benefits generally outweigh the hazards of using ENMs in the oral cavity, and the latter should not prevent the responsible innovation of nanotechnology in dentistry. However, the clinical safety regulations for dental materials have not been specifically updated for ENMs, and some guidance on occupational health for practitioners is also needed. Knowledge gaps for future research include the formation of protein corona in the oral cavity, ENM diffusion through clinically relevant biofilms, and mechanistic investigations on how ENMs strengthen the tooth structure.

Understanding the Particokinetics of Engineered Nanomaterials for Safe and Effective Therapeutic Applications

Increasing numbers of engineered nanomaterials (ENMs) are being developed for therapeutic and diagnostic applications. However, the tunable and varied physicochemical properties of ENMs pose a new challenge for understanding their biological behavior, trafficking, and biodistribution. Herein the concept of "particokinetics" is introduced to address the dynamic biological behavior of ENMs at the molecular level (including gravitational sedimentation, dispersion, aggregation, and interaction with biomolecules in suspending media), cellular level (including cellular uptake, transport, biotransformation, and elimination), and whole-organism level (including absorption, distribution, metabolism, and excretion in vivo). Several mathematical modeling methods are introduced which guide a quantitative description of their biological behavior at different levels. Examples are also provided to delineate the impact of the physicochemical properties of ENMs on their particokinetics. A comprehensive understanding of the in vivo and in vitro particokinetics of ENMs will facilitate the design of tailor-made functional ENMs that act as highly effective and controllable drug-delivery systems with minimal side-effects.

Physicochemical Properties Determine Nanomaterial Cellular Uptake, Transport, and Fate

Although a growing number of innovations have emerged in the fields of nanobiotechnology and nanomedicine, new engineered nanomaterials (ENMs) with novel physicochemical properties are posing novel challenges to understand the full spectrum of interactions at the nano-bio interface. Because these could include potentially hazardous interactions, researchers need a comprehensive understanding of toxicological properties of nanomaterials and their safer design. In depth research is needed to understand how nanomaterial properties influence bioavailability, transport, fate, cellular uptake, and catalysis of injurious biological responses. Toxicity of ENMs differ with their size and surface properties, and those connections hold true across a spectrum of in vitro to in vivo nano-bio interfaces. In addition, the in vitro results provide a basis for modeling the biokinetics and in vivo behavior of ENMs. Nonetheless, we must use caution in interpreting in vitro toxicity results too literally because of dosimetry differences between in vitro and in vivo systems as well the increased complexity of an in vivo environment. In this Account, we describe the impact of ENM physicochemical properties on cellular bioprocessing based on the research performed in our groups. Organic, inorganic, and hybrid ENMs can be produced in various sizes, shapes and surface modifications and a range of tunable compositions that can be dynamically modified under different biological and environmental conditions. Accordingly, we cover how ENM chemical properties such as hydrophobicity and hydrophilicity, material composition, surface functionalization and charge, dispersal state, and adsorption of proteins on the surface determine ENM cellular uptake, intracellular biotransformation, and bioelimination versus bioaccumulation. We review how physical properties such as size, aspect ratio, and surface area of ENMs influence the interactions of these materials with biological systems, thereby affecting their hazard potential. We discuss our actual experimental findings and show how these properties can be tuned to control the uptake, biotransformation, fate, and hazard of ENMs. This Account provides specific information about ENM biological behavior and safety issues. This research also assists the development of safer nanotherapeutics and guides the design of new materials that can execute novel functions at the nano-bio interface.

P019 Perception and reproduction of whole body rotation in standing subjects

Metal-based engineered nanomaterials (ENMs) are known to affect bacterial processes and metabolic activities. While testing their negative effects on biological components, studies traditionally rely on initial exposure concentrations and thereby do not take into consideration the dynamic behavior of ENMs that ultimately determines exposure and toxicity (e.g. ion release). Moreover, functional responses of soil microbial communities to ENMs exposure can be caused by both the particulate forms and the ionic forms, yet their relative contributions remain poorly understood. Therefore, we investigated the dynamic changes of exposure concentrations of three different types of ENMs (nano-ZnO, -Cu and -Pb) and submicron particles (SMPs) in relation to their impact on the capacity of soil bacterial communities to utilize carbon substrates. The different ENMs were chosen to differ in dissolution potential. The dynamic exposures of ENMs were considered using a time weighted average (TWA) approach. The joint toxicity of the particulate forms and the ionic forms of ENMs was evaluated using a response addition model. Our results showed that the effect concentrations of spherical nano-ZnO, -Cu and SMPs, and Pb-based perovskites expressed as TWA were lower than expressed as initial concentrations. Both particulate forms and ionic forms of spherical 18 nm, 43 nm nano-ZnO and 50 nm, 100 nm nano-Cu contribute to the overall response at the EC50 levels. The particulate forms for 150 nm, 200 nm and 900 nm ZnO SMPs and rod-shaped 78 nm nano-Cu mainly affected the soil microbial metabolic potential, while the Cu ions released from spherical 25 nm nano-Cu, 500 nm Cu SMPs and Pb ions released from perovskites mainly described the effects to bacterial communities. Our results indicate that the dynamic exposure of ENMs and relative contributions of particles and ions require consideration in order to pursue a naturally realistic assessment of environmental risks of metal-based ENMs.