Pyrene-polyethylene glycol-modified multi-walled carbon nanotubes: Genotoxicity in V79-4 fibroblast cells.

The genotoxicity of pyrene-polyethylene glycol-modified multi-walled carbon nanotubes (MWCNT-PyPEG), engineered as a nanoplatform for bioapplication, was evaluated. Toxicity was assessed in hamster lung fibroblast cells (V79-4). MTT and Cell Titer Blue methods were used to evaluate cell viability. Genotoxicity was measured by the comet assay and the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay, and fluorescence in situ hybridization (FISH) was used to test induction of structural chromosome aberrations (clastogenic activity) and/or numerical chromosome changes (aneuploidogenic activity). Exogenous metabolic activation enzymes were used in the CBMN-Cyt and FISH tests. Only with metabolic activation, the hybrids caused chromosomal damage, by both clastogenic and aneugenic processes.

Oxidative stress in Ruditapes philippinarum after exposure to different graphene oxide concentrations in the presence and absence of sediment.

The use of carbon nanomaterials (CNMs) is growing in different technological fields, raising concern on their potential impacts on the environment. Given its diverse nanothenological applications, graphene oxide (GO) stands out among the most widely used CNMs. Its hydrophilic capacity enables it to remain stable in suspension in water allowing that GO can be accessible for accumulation by aquatic organisms through ingestion, filtration and superficial dermal contact when present in aquatic ecosystems. Considering that the effects induced to aquatic organisms may depend on environment characteristics, such as temperature, salinity, water pH as well as the presence/absence of sediment, the present study aimed to investigate the influence of sediment on the impacts caused by GO exposure. For this, oxidative stress parameters were measured in the clam Ruditapes philippinarum, exposed to different GO concentrations (0.01, 0.1 and 1 mg/L), in the presence and absence of sediment, for a 28-days experimental period. The results here presented showed that regardless the presence or absence of sediment, most of the biochemical parameters considered were altered when clams were exposed to the highest concentration. The present findings further revealed that in the presence of sediment, clams mostly invested in non-enzymatic defenses (such as reduced glutathione, GSH), while animals exposed to GO in the absence of sediment favored their enzymatic antioxidant defense capacity (catalase, CAT and superoxide dismutase, SOD). This study highlights the relevance of environmental variations as key factors influencing organisms' responses to pollutants.

A carefully designed nanoplatform based on multi walled carbon nanotube wrapped with aptamers.

The interaction between carbon nanotubes (CNTs) and biological molecules of diagnostic and therapeutic interest, as well as the internalization of the CNTs-biomolecules complexes in different types of cell, has been extensively studied due to the potential use of these nanocomplexes as multifunctional nanoplatforms in a great variety of biomedical applications. The effective use of these nanobiotechnologies requires broad multidisciplinary studies of biocompatibility, regarding, for example, the in vitro and in vivo nanotoxicological assays, the capacity to target specific cells and the evaluation of their biomedical potential. However, the first step to be reached is the careful obtainment of the nanoplatform and the understanding of the actual surface composition and structural integrity of the complex system. In this work, we show the detailed construction of a nanoplatform created by the noncovalent interaction between oxidized multi walled carbon nanotubes (MWCNTs) and a DNA aptamer targeting tumor cells. The excess free aptamer was removed by successive washes, revealing the actual surface of the nanocomplex. The MWCNT-aptamer interaction by π-stacking was evidenced and shown to contribute in obtaining a stable nanocomplex compatible with aqueous media having good cell viability. The nucleotide sequence of the aptamer remained intact after the functionalization, allowing its use in further studies of specificity and binding affinity and for the construction of functional nanoplatforms.

Assessment of the effects of graphene exposure in Danio rerio: A molecular, biochemical and histological approach to investigating mechanisms of toxicity.

Graphene has been shown to induce toxicity in mammals and marine crustaceans; however, information regarding oxidative stress in fish is scarce. The aim of this study was to evaluate the mechanism of graphene toxicity in different tissues of Danio rerio, considering different parameters of stress. Animals were injected intraperitoneally (i.p.) with 10 µL of suspensions containing different graphene concentrations (5 and 50 mg/L); the gills, intestine, muscle and brain were analysed 48 h later. There was no significant difference in the expression of the gclc (glutamate cysteine ligase catalytic subunit) and nrf2 (nuclear factor (erythroid-derived 2)-like 2) genes after exposure. In contrast, glutamate cysteine ligase (GCL) and glutathione-S-transferase (GST) activities were modulated and the glutathione (GSH) concentration was reduced in different tissues and at different concentrations. Lipid damage was observed in the gills. Histological analyses were performed to observe if the exposure could induce pathological damage in these tissues. The results showed pathological effects in all tissues, excluding the intestine, after exposure to both concentrations. Overall, these results indicate that graphene induces different grades of toxicological effects that are dependent on the analysed organ, with distinct pathological effects on some and oxidative effects on others. However, the brain and gills seem to be the primary target organs for graphene toxicity.

Exposure to few-layer graphene through diet induces oxidative stress and histological changes in the marine shrimp Litopenaeus vannamei.

The production and use of graphene-based nanomaterials is rapidly increasing. However, few data are available regarding the toxicity of these nanomaterials in aquatic organisms. In the present study, the toxicity of few-layer graphene (FLG) (obtained by chemical exfoliation) was evaluated in different tissues of the shrimp Litopenaeus vannamei following exposure to FLG through a diet for four weeks. Transmission electron microscopy and dynamic light scattering measurements showed a distribution of lateral sheet sizes between 100 and 2000 nm with the average length and width of 800 and 400 nm, respectively. Oxidative stress parameters were analyzed, indicating that FLG exposure led to an increase in the concentration of reactive oxygen species, modulated the activity of antioxidant enzymes such as glutamate cysteine ligase and glutathione-S-transferase, and reduced glutathione levels and total antioxidant capacity. However, the observed modulations were not sufficient to avoid lipid and DNA damage in both gill and hepatopancreas tissues. Furthermore, graphene exposure resulted in morphological changes in hepatopancreas tissues. These results demonstrate that exposure to FLG through the diet induces alterations in the redox state of cells, leading to a subsequent oxidative stress situation. It is therefore clear that nanomaterials presenting these physico-chemical characteristics may be harmful to aquatic biota.