Toxicity and Biotransformation of Carbon-Based Nanomaterials in Marine Microalgae Heterosigma akashiwo.

This work is related to the environmental toxicology risk assessment and evaluation of the possible transformation of carbon-based nanomaterials (CNMs) after contact with marine microalgae. The materials used in the study represent common and widely applied multi-walled carbon nanotubes (CNTs), fullerene (C60), graphene (Gr), and graphene oxide (GrO). The toxicity was evaluated as growth rate inhibition, esterase activity, membrane potential, and reactive oxygen species generation changes. The measurement was performed with flow cytometry after 3, 24, 96 h, and 7 days. The biotransformation of nanomaterials was evaluated after 7 days of microalgae cultivation with CNMs by FTIR and Raman spectroscopy. The calculated toxic level (EC50 in mg/L, 96 h) of used CNMs reduced in the following order: CNTs (18.98) > GrO (76.77) > Gr (159.40) > C60 (414.0). Oxidative stress and membrane depolarization were the main toxic action of CNTs and GrO. At the same time, Gr and C60 decreased the toxic action with time and had no negative impact on microalgae after 7 days of exposure even at the concentration of 125 mg/L. Moreover, C60 and Gr after 7 days of contact with microalgae cells obtained structural deformations.

Uncovering nanotoxicity of a water-soluble and red-fluorescent [70]fullerene nanomaterial.

Engineered fullerene materials have attracted the attention of researchers in the biomedical sciences, especially when their synthetic methodology is developed to endow them with significant levels of water-solubility and bioavailability. In this study, we synthesized and characterized a water-soluble and red-fluorescent [70]fullerene nanomaterial, which fluoresced at 693 nm with a quantum yield of 0.065 and a large Stokes shift (around 300 nm). The fullerene nanomaterial generated mainly singlet oxygen after illumination with blue LED light, while superoxide anion radical production was minimal. The transmission electron microscopy as well as fluorescent studies of Drosophila melanogaster revealed that prepared [70]fullerene nanoparticles had better bioavailability than pristine [70]fullerene nanoparticles. The designed nanomaterials were observed in the apical, perinuclear, and basal regions of digestive cells, as well as the basal lamina of the digestive system's epithelium, with no damage to cell organelles and no activation of degenerative processes and cell death. Our findings provide a new perspective for understanding the in vivo behavior of fullerene nanomaterials and their future application in bioimaging and light-activated nanotherapeutics.

Zinc oxide, titanium dioxide and C60 fullerene nanoparticles, alone and in mixture, differently affect biomarker responses and proteome in the clam Ruditapes philippinarum.

Continuous release of nanoparticles (NPs) into marine coastal environments results in an increased risk of exposure to complex NP mixtures for marine organisms. However, to date, the information on the effects at molecular and biochemical levels induced by the exposure to NPs, singly and as a mixture, is still scant. The present work aimed at exploring the independent and combined effects and the mechanism(s) of action induced by 7-days exposure to 1 µg/L nZnO, 1 µg/L nTiO2 and 1 µg/L FC60 fullerene in the Manila clam Ruditapes philippinarum, using a battery of immunological and oxidative stress biomarkers in haemolymph, gills and digestive gland. In addition, proteomics analyses were performed in gills and the digestive gland, where NP bioaccumulation was also assessed. Increased bioaccumulation of single NPs and the mixture was linked with increased oxidative stress and higher damage to proteins, lipids and DNA in all tissues analysed. The proteomics approach highlighted protein modulation in terms of abundance and damage (higher redox-thiol and carbonylated groups content). In particular, the modulated proteins (16 in gills and 18 in digestive gland) were mostly related to cytoskeleton and energetic metabolism. The digestive gland was the tissue more affected. For all biomarkers measured, increased detrimental effects were observed in the mixture compared to single NP exposures.

The biphasic responses of nanomaterial fullerene on stomatal movement, water status, chlorophyll a fluorescence transient, radical scavenging system and aquaporin-related gene expression in Zea mays under cobalt stress.

Nanomaterial fullerene (FLN) has different responses called the hormesis effect against stress conditions. The favorable/adverse impacts of hormesis on crop quality and productivity are under development in agrotechnology. In this study, the effect of FLN administration (100-250-500mg L-1 for FLN1-2-3, respectively) on growth, water management, gas exchange, chlorophyll fluorescence kinetics and cobalt (Co)-induced oxidative stress in Zea mays was investigated. The negative alterations in relative growth rate (RGR), water status (relative water content, osmotic potential and proline content) and gas exchange/stomatal regulation were removed by FLNs. FLNs were shown to protect photosynthetic apparatus and preserve the photochemistry of photosystems (PSI-PSII) in photosynthesis, chlorophyll fluorescence transients and energy flux damaged under Co stress. The maize leaves exposed to Co stress exhibited a high accumulation of hydrogen peroxide (H2O2) due to insufficient scavenging activity, which was confirmed by reactive oxygen species (ROS)-specific fluorescence visualization in guard cells. FLN regulated the gene expression of ribulose-1,5-bisphosphate carboxylase large subunit (rbcL), nodulin 26-like intrinsic protein1-1 (NIP1-1) and tonoplast intrinsic protein2-1 (TIP2-1) under stress. After stress exposure, FLNs successfully eliminated H2O2 content produced by superoxide dismutase (SOD) activity of catalase (CAT) and peroxidase (POX). The ascorbate (AsA) regeneration was achieved in all FLN applications together with Co stress through the elevated monodehydroascorbate reductase (MDHAR, under all FLNs) and dehydroascorbate reductase (DHAR, only FLN1). However, dose-dependent FLNs (FLN1-2) provided the induced pool of glutathione (GSH) and GSH redox state. Hydroponically applied FLNs removed the restrictions on metabolism and biological process induced by lipid peroxidation (TBARS content) and excessive ROS production. Considering all data, the modulation of treatment practices in terms of FLN concentrations and forms of its application will provide a unique platform for improving agricultural productivity and stress resistance in crops. The current study provided the first findings on the chlorophyll a fluorescence transient and localization of ROS in guard cells of Zea mays exposed to FLN and Co stress.

Fullerene-Based Mimics of Biocatalysts Show Remarkable Activity and Modularity.

The design of catalysts with greater control over catalytic activity and stability is a major challenge with substantial impact on fundamental chemistry and industrial applications. Due to their unparalleled diversity, selectivity, and efficiency, enzymes are promising models for next-generation catalysts, and considerable efforts have been devoted to incorporating the principles of their mechanisms of action into artificial systems. We report a heretofore undocumented catalyst design that introduces fullerenes to the field of biocatalysis, which we refer to as fullerene nanocatalysts, and that emulates enzymatic active sites through multifunctional self-assembled nanostructures. As a proof-of-concept, we mimicked the reactivity of hydrolases using fullerene nanocatalysts functionalized with the basic components of the parent enzyme with remarkable activity. Owing to the versatile amino acid-based functionalization repertoire of fullerene nanocatalysts, these next-generation carbon/biomolecule hybrids have potential to mimic the activity of other families of enzymes and, therefore, offer new perspectives for the design of biocompatible, high-efficiency artificial nanocatalysts.

Toxicity and Antioxidant Activity of Fullerenol C60,70 with Low Number of Oxygen Substituents.

Fullerene is a nanosized carbon structure with potential drug delivery applications. We studied the bioeffects of a water-soluble fullerene derivative, fullerenol, with 10-12 oxygen groups (F10-12); its structure was characterized by IR and XPS spectroscopy. A bioluminescent enzyme system was used to study toxic and antioxidant effects of F10-12 at the enzymatic level. Antioxidant characteristics of F10-12 were revealed in model solutions of organic and inorganic oxidizers. Low-concentration activation of bioluminescence was validated statistically in oxidizer solutions. Toxic and antioxidant characteristics of F10-12 were compared to those of homologous fullerenols with a higher number of oxygen groups:F24-28 and F40-42. No simple dependency was found between the toxic/antioxidant characteristics and the number of oxygen groups on the fullerene's carbon cage. Lower toxicity and higher antioxidant activity of F24-28 were identified and presumptively attributed to its higher solubility. An active role of reactive oxygen species (ROS) in the bioeffects of F10-12 was demonstrated. Correlations between toxic/antioxidant characteristics of F10-12 and ROS content were evaluated. Toxic and antioxidant effects were related to the decrease in ROS content in the enzyme solutions. Our results reveal a complexity of ROS effects in the enzymatic assay system.

Novel pyridinium-type fullerene derivatives as multitargeting inhibitors of HIV-1 reverse transcriptase, HIV-1 protease, and HCV NS5B polymerase.

In the present study, we newly synthesized four types of novel fullerene derivatives: pyridinium/ethyl ester-type derivatives 3b-3l, pyridinium/carboxylic acid-type derivatives 4a, 4e, 4f, pyridinium/amide-type derivative 5a, and pyridinium/2-morpholinone-type derivative 6a. Among the assessed compounds, cis-3c, cis-3d, trans-3e, trans-3h, cis-3l, cis-4e, cis-4f, trans-4f, and cis-5a were found to inhibit HIV-1 reverse transcriptase (HIV-RT), HIV-1 protease (HIV-PR), and HCV NS5B polymerase (HCV NS5B), with IC50 values observed in the micromolar range. Cellular uptake of pyridinium/ethyl ester-type derivatives was higher than that of corresponding pyridinium/carboxylic acid-type derivatives and pyridinium/amide-type derivatives. This result might indicate that pyridinium/ethyl ester-type derivatives are expected to be lead compounds for multitargeting drugs to treat HIV/HCV coinfection.

Blocking effect of fullerene nanoparticles (nC60) on the plant cell structure and its phytotoxicity.

Blockage of nanoparticles on plant pore structures might produce phytotoxicity and affect plant uptake indirectly. This study examined the blocking and phytotoxic effects of fullerene nanoparticles (nC60) on plants at the cellular level. The malondialdehyde content in plant was normal during nC60 exposure, implying that nC60 caused no acute phytotoxicity, while the normalized relative transpiration significantly decreased, showing that the pore structure of roots was seriously blocked by nC60. High power optical microscopy and transmission electron microscope showed that root endothelial cells were squeezed, and inner wall structures were damaged by the extrusion of nanoparticles. Low nC60 concentrations inhibited root uptake of lindane, whereas high nC60 concentrations promoted root uptake of lindane, indicating that serious pore blocking by nC60 damaged root cell structure and hence ready transport of lindane from roots to shoots. Significant alterations of fatty acid (FA) saturation degree of root cell membrane indicated that nC60 led to phytotoxicity in the root cell membrane after long-term exposure and nC60 produced phytotoxicity in the process of blocking root pore structures and interfering with cell membrane fluidity. Moreover, the plant cell structures under phytotoxicity were more likely to be damaged mechanically by the extrusion of nanoparticles. These findings may be helpful to better understand the transport pathways of nanoparticles in plants, the phytotoxicity of nanoparticles and the potential risks of nanomaterials used in agriculture.

Toxicological Aspects of Carbon Nanotubes, Fullerenes and Graphenes.

Nanomedicines exhibit unbelievable capability in overcoming the hurdles faced in biological applications. Carbon nanotubes (CNTs), graphene-family nanomaterials and fullerenes are a class of engineered nanoparticles that have emerged as a new option for possible use in drug/gene delivery for life-threatening diseases. Their adaptability to pharmaceutical applications has opened new vistas for biomedical applications. Successful applications of this family of engineered nanoparticles in various fields may not support their use in medicine due to inconsistent data on toxicity as well as the lack of a centralized toxicity database. Inconsistent toxicological studies and lack of mechanistic understanding have been the reasons for limited understanding of their toxicological aspects. These nanoparticles, when underivatized or pristine, are considered as safe, however less reactive. The derivatized forms or functionalization changes their chemistry significantly to modify their biological effects including toxicity. They can cause acute and long term injuries in tissues by penetration through the the blood-air barrier, blood-alveolus barrier, blood-brain barrier, and blood-placenta barrier. and by accumulating in the lung, liver, and spleen . The toxicological effects are manifested through inflammatory response, DNA damage, apoptosis, autophagy and necrosis. Other factors that largely influence the toxicity of carbon nanotubes, graphenes and fullerenes are the concentration, functionalization, dimensional and surface topographical factors. Thus, a better understanding of the toxicity profile of CNTs, graphene-family nanomaterials and fullerenes in humans, animals and the environment is of significant importance, to improve their biological safety, to facilitate their wide biological application and for the successful commercial application. The exploration of appropriate cell lines to investigate specific receptors and intracellular targets as well as chronic toxicity beyond the proof-of-concept is required.

Antagonistic cytoprotective effects of C60 fullerene nanoparticles in simultaneous exposure to benzo[a]pyrene in a molluscan animal model.

The hypothesis that C60 fullerene nanoparticles (C60) exert an antagonistic interactive effect on the toxicity of benzo[a]pyrene (BaP) has been supported by this investigation. Mussels were exposed to BaP (5, 50 & 100µg/L) and C60 (C60-1mg/L) separately and in combination. Both BaP and C60 were shown to co-localize in the secondary lysosomes of the hepatopancreatic digestive cells in the digestive gland where they reduced lysosomal membrane stability (LMS) or increased membrane permeability, while BaP also induced increased lysosomal lipid and lipofuscin, indicative of oxidative cell injury and autophagic dysfunction. Combinations of BaP and C60 showed antagonistic effects for lysosomal stability, mTORC1 (mechanistic target of rapamycin complex 1) inhibition and intralysosomal lipid (5 & 50µg/L BaP). The biomarker data (i.e., LMS, lysosomal lipidosis and lipofuscin accumulation; lysosomal/cell volume and dephosphorylation of mTORC1) were further analysed using multivariate statistics. Principal component and cluster analysis clearly indicated that BaP on its own was more injurious than in combination with C60. Use of a network model that integrated the biomarker data for the cell pathophysiological processes, indicated that there were significant antagonistic interactions in network complexity (% connectance) at all BaP concentrations for the combined treatments. Loss of lysosomal membrane stability probably causes the release of intralysosomal iron and hydrolases into the cytosol, where iron can generate harmful reactive oxygen species (ROS). It was inferred that this adverse oxidative reaction induced by BaP was ameliorated in the combination treatments by the ROS scavenging property of intralysosomal C60, thus limiting the injury to the lysosomal membrane; and reducing the oxidative damage in the cytosol and to the nuclear DNA. The ROS scavenging by C60, in combination with enhanced autophagic turnover of damaged cell constituents, appeared to have a cytoprotective effect against the toxic reaction to BaP in the combined treatments.

Toxicological effects and bioaccumulation of fullerene C60 (FC60) in the marine bivalve Ruditapes philippinarum.

Fullerene C60 (FC60), with its unique physical properties, has been used in many applications in recent decades. The increased likelihood of direct release into the environment has raised interest in understanding the biological effects of FC60 to aquatic organisms. Nowadays, only few studies have analysed FC60 effects and bioaccumulation in marine organisms following in vivo exposure. To provide new data about FC60 toxicity, Ruditapes philippinarum was selected as target species to assess potential adverse effects of the contaminant. Clams were exposed for 1, 3 and 7 days to predicted environmental concentrations of FC60 (1 and 10 µg/L) and cellular and biochemical responses were evaluated in clams' gills, digestive gland and haemolymph. The FC60 content in gills and digestive gland was determined in all experimental conditions after 7 days of exposure. Results showed an increase in oxidative stress. In particular, a significant modulation in antioxidant enzyme activities, and changes in glutathione S-transferase activity were observed in gills. Moreover, damage to lipids and proteins was detected in FC60-treated (10 µg/L) clams. In digestive gland, slighter variations in antioxidant enzyme activities and damage to molecules were detected. CAT activity was significantly affected throughout the exposure, whereas damage to lipids was evident only at the end of exposure. FC60 accumulation was revealed in both gills and digestive gland, with values up to twelve-fold higher in the latter. Interestingly, haemolymph parameters were slightly affected by FC60 compared to the other tissues investigated. Indeed, only Single Cell Gel Electrophoresis and Neutral Red uptake assays showed increased values in FC60-exposed clams. Moreover, volume and diameter of haemocytes, haemocyte proliferation, and micronucleus assay highlighted significant variations in treated clams, but only in the first phases of exposure, and no changes were detected after 7 days. Our results suggested clam gills as the target tissue for FC60 toxicity under the exposure conditions tested: the high damage detected to lipids and proteins could contribute to long-term problems for the organism.

Acute exposure to C60 fullerene damages pulmonary mitochondrial function and mechanics.

C60 fullerene (C60) nanoparticles, a nanomaterial widely used in technology, can offer risks to humans, overcome biological barriers, and deposit onto the lungs. However, data on its putative pulmonary burden are scanty. Recently, the C60 interaction with mitochondria has been described in vitro and in vivo. We hypothesized that C60 impairs lung mechanics and mitochondrial function. Thirty-five male BALB/c mice were randomly divided into two groups intratracheally instilled with vehicle (0.9% NaCl + 1% Tween 80, CTRL) or C60 (1.0 mg/kg, FUL). Twenty-four hours after exposure, 15 FUL and 8 CTRL mice were anesthetized, paralyzed, and mechanically ventilated for the determination of lung mechanics. After euthanasia, the lungs were removed en bloc at end-expiration for histological processing. Lung tissue elastance and viscance were augmented in FUL group. Increased inflammatory cell number, alveolar collapse, septal thickening, and pulmonary edema were detected. In other six FUL and six CTRL mice, mitochondria expressed reduction in state 1 respiration [FUL = 3.0 ± 1.14 vs. CTRL = 4.46 ± 0.9 (SEM) nmol O2/min/mg protein, p = 0.0210], ATP production (FUL = 122.6 ± 18 vs. CTRL = 154.5 ± 14 µmol/100 µg protein, p = 0.0340), and higher oxygen consumption in state 4 [FUL = 12.56 ± 0.9 vs. CTRL = 8.26 ± 0.6], generation of reactive oxygen species (FUL 733.1 ± 169.32 vs. CTRL = 486.39 ± 73.1 nmol/100 µg protein, p = 0.0313) and reason ROS/ATP [FUL = 8.73 ± 2.3 vs. CTRL = 2.99 ± 0.3]. In conclusion, exposure to fullerene C60 impaired pulmonary mechanics and mitochondrial function, increased ROS concentration, and decrease ATP production.

Eugenol mitigated acute lung but not spermatic toxicity of C60 fullerene emulsion in mice.

C60 fullerene (C60) is a nano-pollutant that can damage the respiratory system. Eugenol exhibits significant anti-inflammatory and antioxidant properties. We aimed to investigate the time course of C60 emulsion-induced pulmonary and spermatic harms, as well as the effect of eugenol on C60 emulsion toxicity. The first group of mice (protocol 1) received intratracheally C60 emulsion (1.0 mg/kg BW) or vehicle and were tested at 12, 24, 72 and 96 h (F groups) thereafter. The second group of mice (protocol 2) received intratracheally C60 emulsion or vehicle, 1 h later were gavaged with eugenol (150 mg/kg) or vehicle, and experiments were done 24 h after instillation. Lung mechanics, morphology, redox markers, cytokines and epididymal spermatozoa were analyzed. Protocol 1: Tissue damping (G) and elastance (H) were significantly higher in F24 than in others groups, except for H in F72. Morphological and inflammatory parameters were worst at 24 h and subsequently declined until 96 h, whereas redox and spermatic parameters worsened over the whole period. Eugenol eliminated the increase in G, H, cellularity, and cytokines, attenuated oxidative stress induced by C60 exposure, but had no effect on sperm. Hence, exposure to C60 emulsion deteriorated lung morphofunctional, redox and inflammatory characteristics and increased the risk of infertility. Furthermore, eugenol avoided those changes, but did not prevent sperm damage.

Deciphering morpho-physiological and phytochemical attributes of Tanacetum parthenium L. plants exposed to C60 fullerene and salicylic acid.

This study was aimed to evaluate the effects of C60 fullerene concentrations (0, 125, 250, 500 and 1000 mg/L) and salicylic acid (0 and 0.2 mM) on growth and phytochemical accumulation of two feverfew genotypes (Pharmasaat and Jelitto) in a factorial experiment based on completely randomized design with three replications. According to the ANOVA, triple interaction of treatments were significant on morphological and phytochmical traits, however, the main effect of treatments only affected physiological attributes. Application of salicylic acid differentially influenced the effects of various concentrations of C60 fullerene on growth traits of both genotypes. In Pharmasaat, foliar application of salicylic acid increased growth traits of plants exposed to C60 fullerene at all concentrations, however, it improved the growth of Jelitto at higher levels of fullerene. The maximum increase of flower + leaf dry weight was recorded at 1000 mg/L C60 fullerene in combination with salicylic acid compared to control for Jelitto. In Pharmasaat, the parthenolide content significantly increased following increase of C60 fullerene up to 250 mg/L with salicylic acid, but a rapid decrease followed at 500-1000 mg/L. SEM images showed a wider deposition (many spheres with different sizes) of C60 fullerene on leaf tissue of Pharmasaat exposed to high concentration, involving changes in trichome density and tissue rupture. The essential oil content was not significantly increased upon experimental treatments compared to control. Based on hierarchical cluster analysis, C60 fullerene and salicylic acid treatments caused to a co-induction of ion leakage, chlorophyll a, essential oil and parthenoloide in Pharmasaat.

Dextran stabilized fullerene soot induced toxicity on alveolar epithelial cells (A549 cells).

Fullerene comprises the major allotrope of carbon holding several fruitful potentials to be applied in various industrial and biomedical scenarios. Scientists have acquired large number of data on fullerene research using its derivatives like C60, C70 etc. Nevertheless, a precise focus on fullerene soot nanopaticles and its toxic impacts in living tissue is still behind mainstay even if it represents the crude parent form of all other derivatives. Present study addresses an acute toxicity profiling of fullerene soot nanoparticles in alveolar epithelial cells (A549) as a paradigm of pulmonary exposure. Surface functionalization was given for fullerene soot nanoparticles using dextran polymer as a mean to establish a stable homogenous dispersion (denoted as dFSNPs hereafter). Following functionalization, dFSNPs were characterized for various parameters including size, surface charge, morphology and functional groups using DLS, Zeta potential analysis, TEM and FT-IR measurements respectively. Effective dextran functionalization was evident from the characteristic peaks in FTIR spectra. Cell viability assessed using MTT and NRU assays; both of which showed a dose dependent cytotoxic response. Thymidine incorporation also confirmed similar trend in viability rate. In accordance with literatures, DCFHDA assay confirmed free radical scavenging activity of fullerene nanoparticles. An altered cellular morphology was observed under fluorescent microscope. Sub-cellular functionalities including lysosomal integrity and mitochondrial stability were found to be compromised at highest tested concentration of dFSNPs (160 µg/ml) without any genotoxic impacts within nuclear premises. FACS analysis following Annexin-PI staining confirmed apoptotic cell death. Hence the overall study substantiated dose dependent toxicity of dFSNPs which is likely to occur during pulmonary exposure.

Resolution of Pulmonary Inflammation Induced by Carbon Nanotubes and Fullerenes in Mice: Role of Macrophage Polarization.

Pulmonary exposure to certain engineered nanomaterials (ENMs) causes chronic lesions like fibrosis and cancer in animal models as a result of unresolved inflammation. Resolution of inflammation involves the time-dependent biosynthesis of lipid mediators (LMs)-in particular, specialized pro-resolving mediators (SPMs). To understand how ENM-induced pulmonary inflammation is resolved, we analyzed the inflammatory and pro-resolving responses to fibrogenic multi-walled carbon nanotubes (MWCNTs, Mitsui-7) and low-toxicity fullerenes (fullerene C60, C60F). Pharyngeal aspiration of MWCNTs at 40 µg/mouse or C60F at a dose above 640 µg/mouse elicited pulmonary effects in B6C3F1 mice. Both ENMs stimulated acute inflammation, predominated by neutrophils, in the lung at day 1, which transitioned to histiocytic inflammation by day 7. By day 28, the lesion in MWCNT-exposed mice progressed to fibrotic granulomas, whereas it remained as alveolar histiocytosis in C60F-exposed mice. Flow cytometric profiling of whole lung lavage (WLL) cells revealed that neutrophil recruitment was the greatest at day 1 and declined to 36.6% of that level in MWCNT- and 16.8% in C60F-treated mice by day 7, and to basal levels by day 28, suggesting a rapid initiation phase and an extended resolution phase. Both ENMs induced high levels of proinflammatory leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) with peaks at day 1, and high levels of SPMs resolvin D1 (RvD1) and E1 (RvE1) with peaks at day 7. MWCNTs and C60F induced time-dependent polarization of M1 macrophages with a peak at day 1 and subsequently of M2 macrophages with a peak at day 7 in the lung, accompanied by elevated levels of type 1 or type 2 cytokines, respectively. M1 macrophages exhibited preferential induction of arachidonate 5-lipoxygenase activating protein (ALOX5AP), whereas M2 macrophages had a high level expression of arachidonate 15-lipoxygenase (ALOX15). Polarization of macrophages in vitro differentially induced ALOX5AP in M1 macrophages or ALOX15 in M2 macrophages resulting in increased preferential biosynthesis of proinflammatory LMs or SPMs. MWCNTs increased the M1- or M2-specific production of LMs accordingly. These findings support a mechanism by which persistent ENM-induced neutrophilic inflammation is actively resolved through time-dependent polarization of macrophages and enhanced biosynthesis of specialized LMs via distinct ALOX pathways.

Effect of pristine fullerene on acquisition, consolidation and retrieval memory in wistar rats.

The present study evaluated the effect of fullerene (C60) under in vitro conditions, in hippocampus homogenates from rats and on the induction of behavioral disabilities. Exposure to in vitro C60 led to an increase in the concentration of reactive oxygen species (ROS) and lipid peroxidation (LPO) of hippocampus treated with of fullerene and suspension. These results indicate that the oxidative stress caused by the exposure to C60 was in part related to an absence of an antioxidant response. In this sense, one-trial inhibitory avoidance task were performed and results showed that fullerene at 0.2 and 0.45 µm impaired the acquisition and consolidation of short and long-term memory. Further, enzymatic analysis in rat hippocampus were not significantly different, however, there was an increase in the content of LPO and ROS produced by fullerene. Overall, the results indicates that fullerene possess neurotoxic properties that impairs behavior and promotes oxidative stress.

Identifying alternative solvents for C60 manufacturing using singular and combined toxicity assessments.

Linseed oil, olive oil, and sunflower oil were selected based on green chemistry principles and C60 solubility as alternative solvents to replace 1,2,4-trimethylbenzene (TMB) for C60 manufacturing. Singular acute toxicity experiments of C60 and the four solvents was performed using Daphnia magna to identify the solvent with the lowest toxicity and estimate the toxicity of C60. The EC50 for C60 was estimated to be higher than 176 ppm. The toxicity of the solvents increased from sunflower oil to olive oil, linseed oil, and TMB. Combined toxicity tests were conducted to investigate the interaction between C60 and the solvent since essential oils can be nanocarriers and facilitate the transport of C60 into the cell membranes, which would increase its toxicity. Various concentrations of C60 (0, 11, 22, 44, 88, and 176 mg/L) were mixed with solvents at their EC50 concentrations. The toxicity of linseed oil increased with increasing C60 concentrations. For olive and sunflower oil, the toxicity was lowered with low concentrations of C60. Olive oil was determined to be a suitable solvent for C60 manufacturing based on singular and combined toxicity assessments. This study showed the importance of considering combined toxicity for solvent selection.

Effects of fullerene C60 in blue mussels: Role of mTOR in autophagy related cellular/tissue alterations.

The effects of C60 on mTOR (mechanistic Target of Rapamycin) activity in mussel digestive gland were investigated. mTOR is a kinase that senses physiological and environmental signals to control eukaryotic cell growth. mTOR is present in two complexes: the phosphorylated mTORC1 regulates cell growth by activating anabolic processes, and by inhibiting catabolic processes (i.e. autophagy); mTORC2 also modulates actin cytoskeleton organization. Mussels were exposed to C60 (0.01, 0.1 and 1 mg/L) for 72 h. Immunocytochemical analysis using a specific antibody revealed the cellular distribution of C60 in mussel digestive gland, already at the lowest concentration. In exposed mussels, the dephosphorylation of mTORC1 and mTORC2 may explain the C60 effects, i.e. the reduction of lysosomal membrane stability, the enhancement of LC3B protein, and the increase of lysosomal/cytoplasmic volume ratio; as well the cytoskeletal alterations. No oxidative stress was observed. Multivariate analysis was used to facilitate the interpretation of the biomarker data. Finally, a low density oligo-microarray was used to understand the cellular responses to fullerene. Transcriptomics identified a number of differentially expressed genes (DEGs) showing a maximum in animals exposed to 0.1 mg/L C60. The most affected processes are associated with energy metabolism, lysosomal activity and cytoskeleton organization. In this study, we report the first data on the subcellular distribution of C60 in mussel's cells; and on the involvement of mTOR inhibition in the alterations due to nanoparticle accumulation. Overall, mTOR deregulation, by affecting protein synthesis, energy metabolism and autophagy, may reduce the capacity of the organisms to effectively grow and reproduce.

Toxicity studies of fullerene C60 (1 µm and 50 nm) administered by nose-only inhalation to Wistar Han [Crl:WI (Han)] rats and B6C3F1/N mice.

Fullerene C60 (C60), a primary allotrope of carbon, is used in a variety of consumer applications including microelectronics, photovoltaics, batteries and fuel cells, and water treatment methods. Human exposure to engineered C60 due to industrial applications may occur via inhalation, oral, dermal, or parenteral routes. In these toxicity and tissue burden studies, male and female Wistar Han rats and B6C3F1/N mice were exposed to fullerene C60 (at least 95% pure) via nose-only inhalation for 3 months. Two different C60 fullerene aggregate sizes, 1 µm diameter (micro-C60) and 50 nm diameter (nano-C60) were studied to assess the potential for differential effects based on particle size. (Abstract Abridged).