Toxicity Of Nanomaterials Research Articles

Impact of Titanium Dioxide-Graphene Oxide (TiO2-GO) Composite Nanoparticle on the Juveniles of the Giant River Prawn, Macrobrachium rosenbergii: Physio-Biochemistry and Transcriptional Response

Nanomaterials are used in many fields, resulting in inevitably releasing into the aquatic environment. The presence of nanomaterials, including TiO2-GO in the aquatic environment, can be toxic to aquatic organisms. However, few studies have focused on the effects of TiO2-GO composite nanoparticle on crustaceans. In the present study, the giant river prawn Macrobrachium rosenbergii juveniles were exposed to two concentrations of TiO2-GO composite nanoparticle (0.1 and 0.5mg/L). The effects of TiO2-GO composite exposure on activities of digestive and antioxidant-related enzymes and expressions of growth and immune-related genes at the transcriptome were studied. The results showed that the survival rate and growth performance were not negatively affected by TiO2-GO composite at the two exposure levels. Nevertheless, exposure to TiO2-GO composite causes an effect on the activities of digestive and antioxidant enzymes in the juvenile prawns. The enzyme activities of CAT, SOD, GSH-Px, AMS, TPS, and LPS in the 0.1mg/L TiO2-GO composite experimental group were markedly reduced than those in the control group. Additionally, the expression level of genes involved in growth and immunity was significantly affected by TiO2-GO composite. After exposure to the 0.1mg/L TiO2-GO composite, the mRNA expression level of MSTN was significantly increased, but the level of EcR, Raptor, and CaBP was significantly decreased. However, the mRNA levels of the CTL, TLR, JAK, and STAT were significantly increased after exposure to the 0.5mg/L concentration of TiO2-GO composite. Furthermore, to understand the molecular mechanism of M. rosenbergii under TiO2-GO composite exposure, RNA-Seq was employed to analyze the changes of the muscle and hepatopancreas transcriptome. Compared with the control group, we identified 5166 and 4784 differentially expressed genes (DEGs) in the muscle and hepatopancreas, respectively (p < 0.05). Based on gene ontology and KEGG analysis, significant differences were observed in the DEGs involved in activity and binding, metabolism, immune response, and environmental information processing. These results showed that exposure to TiO2-GO composite nanoparticle led to the changes of enzyme activity and gene expression, suggesting that TiO2-GO composite existing in aquatic environments would disrupt the physiology of M. rosenbergii. This study will serve as a foundation for subsequent research into the evaluation of nanomaterial toxicity on crustacean species.

Imaging tools for plant nanobiotechnology.

The successful application of nanobiotechnology in biomedicine has greatly changed the traditional way of diagnosis and treating of disease, and is promising for revolutionizing the traditional plant nanobiotechnology. Over the past few years, nanobiotechnology has increasingly expanded into plant research area. Nanomaterials can be designed as vectors for targeted delivery and controlled release of fertilizers, pesticides, herbicides, nucleotides, proteins, etc. Interestingly, nanomaterials with unique physical and chemical properties can directly affect plant growth and development; improve plant resistance to disease and stress; design as sensors in plant biology; and even be used for plant genetic engineering. Similarly, there have been concerns about the potential biological toxicity of nanomaterials. Selecting appropriate characterization methods will help understand how nanomaterials interact with plants and promote advances in plant nanobiotechnology. However, there are relatively few reviews of tools for characterizing nanomaterials in plant nanobiotechnology. In this review, we present relevant imaging tools that have been used in plant nanobiotechnology to monitor nanomaterial migration, interaction with and internalization into plants at three-dimensional lengths. Including: 1) Migration of nanomaterial into plant organs 2) Penetration of nanomaterial into plant tissues (iii)Internalization of nanomaterials by plant cells and interactions with plant subcellular structures. We compare the advantages and disadvantages of current characterization tools and propose future optimal characterization methods for plant nanobiotechnology.

Effects of titanium dioxide (TiO2)/activated carbon (AC) nanoparticle on the growth and immunity of the giant freshwater prawn, Macrobrachium rosenbergii: potential toxicological risks to the aquatic crustaceans.

Due to their unique physicochemical characteristics, nanomaterials exhibit many excellent properties and functions, leading to their applications in numerous fields. The large-scale production and widespread application of nanomaterials have inevitably resulted in their release into the environment, especially the water environment. Several studies have confirmed that exposure to nanomaterials can be toxic to aquatic organisms. However, few studies have focused on the effects of nanomaterial exposure on growth and immunity in crustaceans. In the present study, juvenile Macrobrachium rosenbergii were exposed to different concentrations of titanium dioxide (TiO2)/activated carbon (AC) composite nanomaterial (0.1 and 0.5mg/L) for 45days. The effects of nanoparticle exposure on digestion and antioxidant-related enzyme activities, as well as the expression of growth and immunity-related genes and signaling pathway, were evaluated. Our results show that in response to low concentration of TiO2/AC nanoparticle (0.1mg/L), most of the enzyme activities related to digestion and antioxidation (TPS, LPS, AMS, SOD, and CAT) were diminished. On the contrary, the GSH-Px activity increased under the 0.1mg/L group of TiO2/AC nanoparticle concentration. Additionally, the level of digestive and antioxidant enzyme activities we detected was increased when exposed to 0.5mg/L TiO2/AC nanoparticle. By comparison to the expression level of growth-related genes in the control group, MSTN, CaBP, E75, Raptor, EcR, and EGF were significantly inhibited at 0.1 and 0.5mg/L concentrations of TiO2/AC nanoparticle, whereas the expression level of genes (TLR, JAK, STAT, PPAF, ACP, and AKP) related to immunity was increased when exposed to different concentrations of TiO2/AC nanoparticle. Compared with the control group (0mg/L concentration), 5166 DEGs were identified in the TiO2/AC nanoparticle group, and a large number of DEGs were involved in molting, energy metabolism, stress tolerance, and germ cell development. Moreover, KEGG analysis revealed that many DEGs were assigned into signaling pathways related to metabolic growth and immune stress. These results showed that exposure to TiO2/AC nanoparticle will result in the changes of enzyme activity and routine mRNA expression, suggesting that TiO2/AC nanoparticle which existed in aquatic environment might affect the physiology of M. rosenbergii. This study will provide significant information for the evaluation of nanomaterial toxicity on aquatic crustaceans.

A state-of-the-art review on carbon quantum dots: Prospective, advances, zebrafish biocompatibility and bioimaging in vivo and bibliometric analysis

Carbon quantum dots (CQDs) are recently discovered, in the beginning, a few different types of synthesis are discussed in detail. These include the “top-down” synthetic approach (such as the electrochemical method, laser ablations, and biomass decompositions), the bottom-up approach (such as hydrothermal, microwave, and acidic oxidation), and CQD synthesis methods are derived from waste products. The zebrafish larvae and embryos' fluorescence emission was used to successfully observe the distribution of CQDs throughout the zebrafish life cycle. CQDs used as imaging and molecular probe were shown to have both a “low-toxicity” and “high-biocompatibility” by all the results. Distribution of CQD in zebrafish provided insight into their absorption, distribution, excretion, and metabolism route. The procedures about zebrafish are appropriate for investigating the biocompatibility, adverse effect, toxicity, and transport of nanomaterials, in addition to the screening of drugs using zebrafish as a model. Particular attention is paid to their synthesis, advancement, bibliometric analysis, and technical applications of bioimaging. The bibliometric analysis of title search results from 2011 to 2022 for keywords, nations; co-author networks, co-occurrence, and journals are visualizations using the Scopus database and RStudio software described. Data were extracted from a total of 1456 papers, 379 review articles, and 1377 article proceedings within 12 years (2011−2022).

Hemolytic Activity of Nanoparticles as a Marker of Their Hemocompatibility.

The potential use of nanomaterials in medicine offers opportunities for novel therapeutic approaches to treating complex disorders. For that reason, a new branch of science, named nanotoxicology, which aims to study the dangerous effects of nanomaterials on human health and on the environment, has recently emerged. However, the toxicity and risk associated with nanomaterials are unclear or not completely understood. The development of an adequate experimental strategy for assessing the toxicity of nanomaterials may include a rapid/express method that will reliably, quickly, and cheaply make an initial assessment. One possibility is the characterization of the hemocompatibility of nanomaterials, which includes their hemolytic activity as a marker. In this review, we consider various factors affecting the hemolytic activity of nanomaterials and draw the reader's attention to the fact that the formation of a protein corona around a nanoparticle can significantly change its interaction with the red cell. This leads us to suggest that the nanomaterial hemolytic activity in the buffer does not reflect the situation in the blood plasma. As a recommendation, we propose studying the hemocompatibility of nanomaterials under more physiologically relevant conditions, in the presence of plasma proteins in the medium and under mechanical stress.

Potential Environmental and Health Implications from the Scaled-Up Production and Disposal of Nanomaterials Used in Biosensors.

Biosensors often combine biological recognition elements with nanomaterials of varying compositions and dimensions to facilitate or enhance the operating mechanism of the device. While incorporating nanomaterials is beneficial to developing high-performance biosensors, at the stages of scale-up and disposal, it may lead to the unmanaged release of toxic nanomaterials. Here we attempt to foster connections between the domains of biosensors development and human and environmental toxicology to encourage a holistic approach to the development and scale-up of biosensors. We begin by exploring the toxicity of nanomaterials commonly used in biosensor design. From our analysis, we introduce five factors with a role in nanotoxicity that should be considered at the biosensor development stages to better manage toxicity. Finally, we contextualize the discussion by presenting the relevant stages and routes of exposure in the biosensor life cycle. Our review found little consensus on how the factors presented govern nanomaterial toxicity, especially in composite and alloyed nanomaterials. To bridge the current gap in understanding and mitigate the risks of uncontrolled nanomaterial release, we advocate for greater collaboration through a precautionary One Health approach to future development and a movement towards a circular approach to biosensor use and disposal.

Environmental Hazards of Nanobiomaterials (Hydroxyapatite-Based NMs)-A Case Study with Folsomia candida-Effects from Long Term Exposure.

Hydroxyapatite (HA) is a calcium phosphate used in many fields, including biomedical applications. In particular, ion-doped HA nanomaterials (nHA) are developed for their increased bioactivity, particularly in the fields of regenerative medicine and nanomedicine. In this study, we assessed the ecotoxicological impact of five nHA materials: a synthesized calcium hydroxyapatite (CaP-HA), superparamagnetic iron-doped hydroxyapatite (Fe-HA), titanium-doped hydroxyapatite (Ti-HA), alginate/titanium-doped hydroxyapatite hybrid composite (Ti-HA-Alg), and a commercial HA. The soil ecotoxicology model species Folsomia candida (Collembola) was used, and besides the standard reproduction test (28 days), an extension to the standard for one more generation was performed (56 days). Assessed endpoints included the standard survival and reproduction, and additionally, growth. Exposure via the standard (28 days) did not cause toxicity, but reproduction increased in commercial HA (significantly at 320 mg HA/kg) whereas via the extension (56 days) it decreased in all tested concentrations. Juveniles' size (56 days) was reduced in all tested nHA materials, except commercial HA. nHA materials seem to trigger a compromise between reproduction and growth. Long-term effects could not be predicted based on the standard shorter exposure; hence, the testing of at least two generations (56 days) is recommended to assess the toxicity of nanomaterials, particularly in F. candida. Further, we found that the inclusion of size as additional endpoint is highly relevant.

Effect of Secondary Structures on the Adsorption of Peptides onto Hydrophobic Solid Surfaces Revealed by SALDI-TOF and MD Simulations.

The adsorption of peptides and proteins on hydrophobic solid surfaces has received considerable research attention owing to their wide applications to biocompatible nanomaterials and nanodevices, such as biosensors and cell adhesion materials with reduced nanomaterial toxicity. However, fundamental understandings about physicochemical hydrophobic interactions between peptides and hydrophobic solid surfaces are still unknown. In this study, we investigate the effect of secondary structures on adsorption energies between peptides and hydrophobic solid surfaces via experimental and theoretical analyses using surface-assisted laser desorption/ionization-time-of-flight (SALDI-TOF) and molecular dynamics (MD) simulations. The hydrophobic interactions between peptides and hydrophobic solid surfaces measured via SALDI-TOF and MD simulations indicate that the hydrophobic interaction of peptides with random coil structures increased more than that of peptides with an α-helix structure when polar amino acids are replaced with hydrophobic amino acids. Additionally, our study sheds new light on the fundamental understanding of the hydrophobic interaction between hydrophobic solid surfaces and peptides that have diverse secondary structures.

An Evaluation of the Biocatalyst for the Synthesis and Application of Zinc Oxide Nanoparticles for Water Remediation—A Review

Global water scarcity is threatening the lives of humans, and it is exacerbated by the contamination of water, which occurs because of increased industrialization and soaring population density. The available conventional physical and chemical water treatment techniques are hazardous to living organisms and are not environmentally friendly, as toxic chemical elements are used during these processes. Nanotechnology has presented a possible way in which to solve these issues by using unique materials with desirable properties. Zinc oxide nanoparticles (ZnO NPs) can be used effectively and efficiently for water treatment, along with other nanotechnologies. Owing to rising concerns regarding the environmental unfriendliness and toxicity of nanomaterials, ZnO NPs have recently been synthesized through biologically available and replenishable sources using a green chemistry or green synthesis protocol. The green-synthesized ZnO NPs are less toxic, more eco-friendly, and more biocompatible than other chemically and physically synthesized materials. In this article, the biogenic synthesis and characterization techniques of ZnO NPs using plants, bacteria, fungi, algae, and biological derivatives are reviewed and discussed. The applications of the biologically prepared ZnO NPs, when used for water treatment, are outlined. Additionally, their mechanisms of action, such as the photocatalytic degradation of dyes, the production of reactive oxygen species (ROS), the generation of compounds such as hydrogen peroxide and superoxide, Zn2+ release to degrade microbes, as well as their adsorbent properties with regard to heavy metals and other contaminants in water bodies, are explained. Furthermore, challenges facing the green synthesis of these nanomaterials are outlined. Future research should focus on how nanomaterials should reach the commercialization stage, and suggestions as to how this ought to be achieved are presented.

Current Scenario of MXene-Based Nanomaterials for Wastewater Remediation: A Review

Rapid urban and industrial sectors generate massive amounts of wastewater, creating severe ecological disruption and harming living organisms. The number of harmful pollutants such as dyes, heavy metals, antibiotics, phenolic compounds, and volatile and several organic chemicals discharged into aquatic systems varies depending on the effluent composition of various sectors. MXene-based composites with unique characteristics were spotlighted as newly developed nanomaterials specifically for environmental-related applications. Therefore, this review broadly discusses the properties, basic principles of MXene, and synthesis routes for developing different MXene-based nanomaterials. The most current strategies on the energy and environmental applications of MXene-based nanomaterials, particularly in photocatalysis, adsorption, and water splitting, were deeply explored for the remediation of different pollutants and hydrogen (H2) evolution from wastewater. The detailed mechanism for H2 evolution and the remediation of industrial pollutants via photocatalysis and adsorption processes was elaborated. The multi-roles of MXene-based nanomaterials with their regeneration possibilities were emphasized. Several essential aspects, including the economic, toxicity and ecological power of MXene-based nanomaterials, were also discussed regarding their opportunity for industrialization. Finally, the perspectives and challenges behind newly developed MXene and MXene-based nanomaterials for environmental pollution were reviewed.

The potential for nanomaterial toxicity affecting the male reproductive system.

With the development of nanotechnology, nanomaterials offer great advantages in a wide variety of industrial and consumer products, and show promise for biomedical applications. However, with these new products, nanomaterial pollutants may enter the human body to cause adverse health effects, including hazards to the male reproductive system. Nanomaterials can enter the body through inhalation, oral exposure, or intravenous injection, and reach the testis via the blood, penetrate the Sertoli cell barrier, and directly or indirectly elicit toxicopathological changes to the testicles. These may then trigger hormone disorders, inhibit spermatogenic cell proliferation, and induce apoptosis, ultimately leading to a decrease in sperm motility and number, ultimately diminishing male reproductive capacity. This review will discuss the toxicological effects of nanomaterials on the male reproductive system, including inflammation, the impact on the hypothalamic-pituitary-gonadal axis (HPG axis), lipid peroxidation, and free ion release relevant to germ cells, Sertoli cell tight junctions, and the gonadal endocrine system. This article is categorized under: Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials.

Nanotechnology in Residential Building Materials for Better Fire Protection and Life Safety Outcomes

Residential fires are the main source of fire deaths and injuries both in the United States and globally. As such, better fire-resistant building materials are needed to bolster fire protection and to enhance life safety. This is during a time when fewer materials are being used to construct homes. Nanotechnology may be a solution if it can overcome its current barriers to widespread adoption in residential construction, namely economy, sustainability, and safety. This research effort includes a critical examination of the literature from a safety perspective to address fire deaths and prevent personal injuries and illnesses by targeting fortification of residential construction building materials via the use of nanotechnology. The paper reviews nanotechnology for building materials by material type, known toxicity of various nanomaterials used in construction, and a discussion on a way forward through assessing materials by their ability to satisfy the requirements of sustainability, economy, and safety- both as a material designed to reduce fire injury and death and from a toxicological hazard perspective.

Proteomic profiling reveals mitochondrial toxicity of nanosilver and silver nitrate in the gill of common carp

Mitochondria are recognized as an important target organelle for the toxicity of nanomaterials. Although the toxic effects of silver nanoparticles (AgNPs) on mitochondria have been widely reported, the mechanism behind the toxicity remains unclear. In this study, the effects of two forms of silver (AgNPs and AgNO3) on carp gill mitochondria were investigated by analyzing the mitochondrial ultrastructure, physicochemical properties of mitochondrial membrane, and mitochondrial proteomics. After exposure of common carp to AgNPs (0.75 mg/L) and AgNO3 (0.05 mg/L) for 96 h, both forms of silver were shown to cause gill mitochondrial lesions, including irregular shape, loss of mitochondrial cristae, and increased mitochondrial membrane permeability. Proteomics results revealed that AgNPs and AgNO3 induced 362 and 297 differentially expressed proteins (DEPs) in gill mitochondria, respectively. Among the DEPs, 244 were shared between AgNPs and AgNO3 treatments. These shared proteins were mainly distributed in the mitochondrial membrane and matrix, and were significantly enriched in the tricarboxylic acid (TCA) cycle and oxidative phosphorylation pathway. The functional annotation of DEPs induced by both silver forms was mainly involved in energy production and conversion. These results indicated that the toxic mechanism of AgNPs and AgNO3 on gill mitochondria were comparable and the two forms of silver caused mitochondrial dysfunction in fish gills by inhibiting the TCA cycle and disrupting the electron transport chain.

Extracellular vesicles as a novel photosensitive drug delivery system for enhanced photodynamic therapy.

Photodynamic therapy (PDT) is a promising non-invasive therapeutic approach that utilizes photosensitizers (PSs) to generate highly reactive oxygen species (ROS), including singlet oxygen, for removal of targeted cells. PDT has been proven efficacious for the treatment of several diseases, including cancer, cardiovascular disease, inflammatory bowel disease, and diabetic ocular disease. However, the therapeutic efficacy of PDT is limited and often accompanied by side effects, largely due to non-specific delivery of PSs beyond the desired lesion site. Over the past decade, despite various nanoparticular drug delivery systems developed have markedly improved the treatment efficacy while reducing the off-target effects of PSs, concerns over the safety and toxicity of synthetic nanomaterials following intravenous administration are raised. Extracellular vesicles (EVs), a type of nanoparticle released from cells, are emerging as a natural drug delivery system for PSs in light of EV’s potentially low immunogenicity and biocompatibility compared with other nanoparticles. This review aims to provide an overview of the research progress in PS delivery systems and propose EVs as an alternative PS delivery system for PDT. Moreover, the challenges and future perspectives of EVs for PS delivery are discussed.

High throughput embryonic zebrafish test with automated dechorionation to evaluate nanomaterial toxicity

Engineered nanomaterials pose occupational health and environmental concerns as they possess unique physical and chemical properties that can contribute to toxicity. High throughput toxicity screening methods are needed to address the increasing number of nanomaterials in production. Here we used a zebrafish photomotor response (PMR) test to evaluate a set of fifteen nanomaterials with military relevance. Automated dechorionation of zebrafish embryos was used to enhance nanomaterials bioavailability. Optimal PMR activity in zebrafish embryos was found at 30–31 hours post-fertilization (hpf). Behavioral and toxicological responses were measured at 30 and 120 hpf; behavioral responses were found for thirteen of the fifteen nanomaterials and acute toxicity (LC50) levels for nine of the fifteen nanomaterials below the maximum test concentration of 500 μg/ml. Physico-chemical characterization of the nanomaterials detected endotoxin and bacterial contamination in two of the tested samples, which may have contributed to observed toxicity and reinforces the need for physical and chemical characterization of nanomaterials use in toxicity testing. The zebrafish PMR test, together with automated dechorionation, provides an initial rapid assessment of the behavioral effects and toxicity of engineered nanomaterials that can be followed up by physico-chemical characterization if toxicity is detected, reducing the amount of time and monetary constraints of physico-chemical testing.

Enhanced photodegradation of rifampicin and co-trimoxazole by ZnO/ZnMn2O4/ZnS-PVA and its genotoxicity studies on Allium cepa

Oxygen vacancies and its associated defect states have a great influence on the electronic and structural aspects of semiconductor photocatalysts, yet there is paucity of investigations about the influence of the defect states on their photocatalytic properties. Herein, this study reports the hierarchical fabrication of oxygen vacancy enriched ZnO/ZnMn2O4/ZnS-PVA nanocomposite (NCs) for the enhanced photodegradation of rifampicin and co-trimoxazole. The formation of lattice expansion induced oxygen vacancies and its associated Urbach tail energy, and n-p-n heterojunction-based S-scheme charge transfer path synergistically contributed to the boosted photocatalytic performance of the as prepared NCs. The photocatalytic performance of the nanomaterial towards rifampicin and co-trimoxazole has been determined to be 80% and 90% under visible light irradiation, respectively. Furthermore, various operating parameters including the concentration of NCs and drug, pH and interference of various ions have been evaluated. The degraded product intermediates have been elucidated by GC-MS analysis. The toxicity of the as-prepared nanomaterials has been evaluated by treating the samples with root tips of Allium cepa, where the NCs was found to be non-toxic. The study provides a new-fangled insight on the preparation and fabrication of non-toxic and defect rich nanomaterials which may help stimulate this area of research.

Nanomaterials for biogas augmentation towards renewable and sustainable energy production: A critical review.

Nanotechnology is considered one of the most significant advancements in science and technology over the last few decades. However, the contemporary use of nanomaterials in bioenergy production is very deficient. This study evaluates the application of nanomaterials for biogas production from different kinds of waste. A state-of-the-art comprehensive review is carried out to elaborate on the deployment of different categories of nano-additives (metal oxides, zero-valent metals, various compounds, carbon-based nanomaterials, nano-composites, and nano-ash) in several kinds of biodegradable waste, including cattle manure, wastewater sludge, municipal solid waste, lake sediments, and sanitary landfills. This study discusses the pros and cons of nano-additives on biogas production from the anaerobic digestion process. Several all-inclusive tables are presented to appraise the literature on different nanomaterials used for biogas production from biomass. Future perspectives to increase biogas production via nano-additives are presented, and the conclusion is drawn on the productivity of biogas based on various nanomaterials. A qualitative review of relevant literature published in the last 50 years is conducted using the bibliometric technique for the first time in literature. About 14,000 research articles are included in this analysis, indexed on the Web of Science. The analysis revealed that the last decade (2010–20) was the golden era for biogas literature, as 84.4% of total publications were published in this timeline. Moreover, it was observed that nanomaterials had revolutionized the field of anaerobic digestion, methane production, and waste activated sludge; and are currently the central pivot of the research community. The toxicity of nanomaterials adversely affects anaerobic bacteria; therefore, using bioactive nanomaterials is emerging as the best alternative. Conducting optimization studies by varying substrate and nanomaterials’ size, concentration and shape is still a field. Furthermore, collecting and disposing nanomaterials at the end of the anaerobic process is a critical environmental challenge to technology implementation that needs to be addressed before the nanomaterials assisted anaerobic process could pave its path to the large-scale industrial sector.

LP-70 Toxicity of graphene nanomaterials in HEK 293T cells – effects of surface modifications by thermal treatment

LP-70 Toxicity of graphene nanomaterials in HEK 293T cells – effects of surface modifications by thermal treatment

Occurrence, transport, and toxicity of nanomaterials in soil ecosystems: a review

Occurrence, transport, and toxicity of nanomaterials in soil ecosystems: a review

Interlaboratory evaluation of a digital holographic microscopy–based assay for label-free in vitro cytotoxicity testing of polymeric nanocarriers

State-of-the-art in vitro test systems for nanomaterial toxicity assessment are based on dyes and several staining steps which can be affected by nanomaterial interference. Digital holographic microscopy (DHM), an interferometry-based variant of quantitative phase imaging (QPI), facilitates reliable proliferation quantification of native cell populations and the extraction of morphological features in a fast and label- and interference-free manner by biophysical parameters. DHM therefore has been identified as versatile tool for cytotoxicity testing in biomedical nanotechnology. In a comparative study performed at two collaborating laboratories, we investigated the interlaboratory variability and performance of DHM in nanomaterial toxicity testing, utilizing complementary standard operating procedures (SOPs). Two identical custom-built off-axis DHM systems, developed for usage in biomedical laboratories, equipped with stage-top incubation chambers were applied at different locations in Europe. Temporal dry mass development, 12-h dry mass increments and morphology changes of A549 human lung epithelial cell populations upon incubation with two variants of poly(alkyl cyanoacrylate) (PACA) nanoparticles were observed in comparison to digitonin and cell culture medium controls. Digitonin as cytotoxicity control, as well as empty and cabazitaxel-loaded PACA nanocarriers, similarly impacted 12-h dry mass development and increments as well as morphology of A549 cells at both participating laboratories. The obtained DHM data reflected the cytotoxic potential of the tested nanomaterials and are in agreement with corresponding literature on biophysical and chemical assays. Our results confirm DHM as label-free cytotoxicity assay for polymeric nanocarriers as well as the repeatability and reproducibility of the technology. In summary, the evaluated DHM assay could be efficiently implemented at different locations and facilitates interlaboratory in vitro toxicity testing of nanoparticles with prospects for application in regulatory science.Graphical abstract