Toxicity Of Carbon Nanomaterials Research Articles

Toxicity of Carbon Nanomaterials-Towards Reliable Viability Assessment via New Approach in Flow Cytometry.

The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful effects of carbon nanomaterials. Even though researchers already reported the interference of carbon nanomaterials with common toxicity assays, there is still, unfortunately, a large number of studies that neglect this fact. In this study, we investigated interference of four bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C3N4) and carbon dots (QCDs)) in commonly used LIVE/DEAD assay. When a standard procedure was applied, materials caused various types of interference. While positively charged g-C3N4 and QCDs induced false results through the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false signals due to the complex quenching effect of the fluorescent dye of a LIVE/DEAD kit. Thus, we developed a new approach using a specific gating strategy based on additional controls that successfully overcame all types of interference and lead to reliable results in LIVE/DEAD assay. We suggest that the newly developed procedure should be a mandatory tool for all in vitro flow cytometry assays of any class of carbon nanomaterials.

Similar toxicity mechanisms between graphene oxide and oxidized multi-walled carbon nanotubes in Microcystis aeruginosa

In photosynthetic microorganisms, the toxicity of carbon nanomaterials (CNMs) is typically characterized by a decrease in growth, viability, photosynthesis, as well as the induction of oxidative stress. However, it is currently unclear how the shape of the carbon structure in CNMs, such as in the 1-dimensional carbon nanotubes (CNTs) compared to the two-dimensional graphene oxide (GO), affects the way they interact with cells. In this study, the effects of GO and oxidized multi-walled CNTs were compared in the cyanobacterium Microcystis aeruginosa to determine the similarities or differences in how the two CNMs interact with and induce toxicity to cyanobacteria. Using change in Chlorophyll a concentrations, the effective concentrations inducing 50% inhibition (EC50) at 96 h are found to be 11.1 μg/mL and 7.38 μg/mL for GO and CNTs, respectively. The EC50 of the two CNMs were not found to be statistically different. Changes in fluorescein diacetate and 2′,7′-dichlorodihydrofluorescein diacetate fluorescence, measured at the EC50 concentrations, suggest a decrease in esterase enzyme activity but no oxidative stress. Scanning and transmission electron microscopy imaging did not show extensive membrane damage in cells exposed to GO or CNTs. Altogether, the decrease in metabolic activity and photosynthetic activity without oxidative stress or membrane damage support the hypothesis that both GO and CNTs induced indirect toxicity through physical mechanisms associated with light shading and cell aggregation. This indirect toxicity explains why the intrinsic differences in shape, size, and surface properties between CNTs and GO did not result in differences in how they induce toxicity to cyanobacteria.

Toxicity of Carbon Nanomaterials and Their Potential Application as Drug Delivery Systems: In Vitro Studies in Caco-2 and MCF-7 Cell Lines.

Carbon nanomaterials have attracted increasing attention in biomedicine recently to be used as drug nanocarriers suitable for medical treatments, due to their large surface area, high cellular internalization and preferential tumor accumulation, that enable these nanomaterials to transport chemotherapeutic agents preferentially to tumor sites, thereby reducing drug toxic side effects. However, there are widespread concerns on the inherent cytotoxicity of carbon nanomaterials, which remains controversial to this day, with studies demonstrating conflicting results. We investigated here in vitro toxicity of various carbon nanomaterials in human epithelial colorectal adenocarcinoma (Caco-2) cells and human breast adenocarcinoma (MCF-7) cells. Carbon nanohorns (CNH), carbon nanotubes (CNT), carbon nanoplatelets (CNP), graphene oxide (GO), reduced graphene oxide (GO) and nanodiamonds (ND) were systematically compared, using Pluronic F-127 dispersant. Cell viability after carbon nanomaterial treatment followed the order CNP < CNH < RGO < CNT < GO < ND, being the effect more pronounced on the more rapidly dividing Caco-2 cells. CNP produced remarkably high reactive oxygen species (ROS) levels. Furthermore, the potential of these materials as nanocarriers in the field of drug delivery of doxorubicin and camptothecin anticancer drugs was also compared. In all cases the carbon nanomaterial/drug complexes resulted in improved anticancer activity compared to that of the free drug, being the efficiency largely dependent of the carbon nanomaterial hydrophobicity and surface chemistry. These fundamental studies are of paramount importance as screening and risk-to-benefit assessment towards the development of smart carbon nanomaterial-based nanocarriers.

Influence of functional groups on toxicity of carbon nanomaterials

Abstract. It has been recognized that carbon nanomaterials and soot particles are toxic for human health, but the influence of functionalization on their toxicity as well as the evolution of the toxicity of carbon nanomaterials due to chemical aging in the atmosphere is still controversial. In the current study, the oxidation potential measured by dithiothreitol (DTT) decay rate and the cytotoxicity to murine macrophage cells of different functionalized carbon nanomaterials were investigated to understand the role of functionalization in their toxicities. The DTT decay rates of special black 4A (SB4A), graphene, graphene oxide, single-walled carbon nanotubes (SWCNTs), SWCNT-OH and SWCNT-COOH were 45.9±3.0, 58.5±6.6, 160.7±21.7, 38.9±8.9, 57.0±7.2 and 36.7±0.2 pmol min−1 µg−1, respectively. Epoxide was found to be mainly responsible for the highest DTT decay rate of graphene oxide compared to other carbon nanomaterials based on comprehensive characterizations. Both carboxylation and hydroxylation showed little influence on the oxidation potential of carbon nanomaterials, while epoxidation contributes to the enhancement of oxidation potential. All these carbon nanomaterials were toxic to the murine J774 cell line. However, oxidized carbon nanomaterials (graphene oxide, SWCNT-OH and SWCNT-COOH) showed weaker cytotoxicity to the J774 cell line compared to the corresponding control sample as far as the metabolic activity was considered and stronger cytotoxicity to the J774 cell line regarding the membrane integrity and DNA incorporation. These results imply that epoxidation might enhance the oxidation potential of carbon nanomaterials.

Study of MWNTS Influence upon Liver Histological and Histochemical Parameters in Laboratory Mice: Preliminary Results

Preliminary evaluation of toxic effect of commercially manufactured carbon nanostructured material based on multi-walled carbon nanotubes (MWCNT) upon laboratory mice C57B6/DBA2 males was carried out. It was found that thirty-day oral administration of nanotubes in doses of 0.3 and 3 mg/kg has no effect on liver condition, while administration of 30 mg/kg leads to formation of inflammatory infiltrates together with hepatocyte structure modification. The obtained results are of potential interest for development of industrial safety standards in nanomaterial handling and for development of standards for reproductive toxicity of carbon nanomaterials.

Carbon nanomaterials in clean and contaminated soils: environmental implications and applications

Abstract. The exceptional sorptive ability of carbon nanomaterials (CNMs) for hydrophobic organic contaminants (HOCs) is driven by their characteristically large reactive surface areas and highly hydrophobic nature. Given these properties, it is possible for CNMs to impact on the persistence, mobility and bioavailability of contaminants within soils, either favourably through sorption and sequestration, hence reducing their bioavailability, or unfavourably through increasing contaminant dispersal. This review considers the complex and dynamic nature of both soil and CNM physicochemical properties to determine their fate and behaviour, together with their interaction with contaminants and the soil microflora. It is argued that assessment of CNMs within soil should be conducted on a case-by-case basis and further work to assess the long-term stability and toxicity of sorbed contaminants, as well as the toxicity of CNMs themselves, is required before their sorptive abilities can be applied to remedy environmental issues.

Quantification of Carbon Nanomaterials in Vivo

A diverse array of carbon nanomaterials (NMs), including fullerene, carbon nanotubes (CNTs), graphene, nanodiamonds, and carbon nanoparticles, have been discovered and widely applied in a variety of industries. Carbon NMs have been detected in the environment and have a strong possibility of entering the human body. The safety of carbon NMs has thus become a serious concern in academia and society. To achieve strict biosafety assessments, researchers need to fully understand the effects and fates of NMs in the human body, including information about absorption, distribution, metabolism, excretion, and toxicity (ADME/T). To acquire the ADME data, researchers must quantify NMs, but carbon NMs are very difficult to quantify in vivo. The carbon background in a typical biological system is high, particularly compared with the much lower concentration of carbon NMs. Moreover, carbon NMs lack a specific detection signal. Therefore, isotopic labeling, with its high sensitivity and specificity, is the first choice to quantify carbon NMs in vivo. Previously, researchers have used many isotopes, including ¹³C, ¹⁴C, ¹²⁵I, ¹³¹I, ³H, ⁶⁴Cu, ¹¹¹In, ⁸⁶Y, 99mTc, and ⁶⁷Ga, to label carbon NMs. We used these isotopic labeling methods to study the ADME of carbon NMs via different exposure pathways in animal models. Except for the metabolism of carbon NMs, which has seldom been investigated, significant amounts of data have been reported on the in vivo absorption, distribution, excretion, and toxicity of carbon NMs, which have revealed characteristic behaviors of carbon NMs, such as reticuloendothelial system (RES) capture. However, the complexity of the biological systems and diverse preparation and functionalization of the same carbon NMs have led to inconsistent results across different studies. Therefore, the data obtained so far have not provided a compatible and systematic profile of biosafety. Further efforts are needed to address these problems. In this Account, we review the in vivo quantification methods of carbon NMs, focusing on isotopic labeling and tracing methods, and summarize the related labeling, purification, bio-sampling, and detection of carbon NMs. We also address the advantages, applicable situations, and limits of various labeling and tracing methods and propose guidelines for choosing suitable labeling methods. A collective analysis of the ADME information on various carbon NMs in vivo would provide general principles for understanding the fate of carbon NMs and the effects of chemical functionalization and aggregation of carbon NMs on their ADME/T in vivo and their implications in nanotoxicology and biosafety evaluations.

Effects of carbon nanomaterials fullerene C60 and fullerol C60(OH)18–22 on gills of fish Cyprinus carpio (Cyprinidae) exposed to ultraviolet radiation

In consequence of their growing use and demand, the inevitable environmental presence of nanomaterials (NMs) has raised concerns about their potential deleterious effects to aquatic environments. The carbon NM fullerene (C60), which forms colloidal aggregates in water, and its water-soluble derivative fullerol (C60(OH)18–22), which possesses antioxidant properties, are known to be photo-excited by ultraviolet (UV) or visible light. To investigate their potential hazards to aquatic organisms upon exposure to UV sunlight, this study analyzed (a) the in vitro behavior of fullerene and fullerol against peroxyl radicals (ROO) under UV-A radiation and (b) the effects of these photo-excited NMs on oxidative stress parameters in functional gills extracted from the fish Cyprinus carpio (Cyprinidae). The variables measured were the total antioxidant capacity, lipid peroxidation (TBARS), the activities of the antioxidant enzymes glutathione reductase (GR) and glutamate cysteine ligase (GCL), and the levels of the non-enzymatic antioxidant glutathione (GSH). The obtained results revealed the following: (1) both NMs behaved in vitro as antioxidants against ROO in the dark and as pro-oxidants in presence of UV-A, the latter effect being reversed by the addition of sodium azide, which is a singlet oxygen (1O2) quencher; (2) fullerene induced toxicity with or without UV-A incidence, with a significant (p<0.05) increase in lipid peroxidation (with greater damage under illumination), a decrease in GCL activity, and the depletion of GSH stocks (under illumination), all of which were attributed to 1O2 generation; and (3) fullerol also decreased GCL activity and GSH formation (p<0.05) but without lipid damage. The overall results show that fullerene can be toxic with or without light incidence, whereas UV radiation seems to play a key role in the environmental toxicity of carbon NMs through 1O2 formation.

Acute toxicity of carbon-based nanomaterials to Escherichia coli is partially dependent on the presence of process impurities

Using inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectroscopy, we perform a compositional analysis of ten commercially available and laboratory carbon nanomaterials (CNMs). It is found that, depending on the type of CNMs and applied purification technology, they can contain 20 to 50 out of the 62 identified chemical elements, totally amounting to 0.03 to 23.1% of the weight of the sample under analysis. It is shown that the acute toxicity of CNMs, which is determined according to the effect on the luminescence intensity of a sensor Escherichia coli strain carrying cloned luxCDABE genes of Photobacterium leiognathi, is partially dependent on the presence of process impurities of Ni, Y, Zn, Sm, Gd, Ho, and Fe. With an increase in the time of contact between the sensor microorganism and the CNM, the latter develop intrinsic biotoxicity, which decreases the significance of the impurity composition in determining this activity within the dynamics of the experiment.