M. V. B. Pinheiro

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.

Synthesis of C60(OH)18-20 in aqueous alkaline solution under O2-atmosphere

In this work we report on an alternative synthesis of water-soluble fullerenes known as fullerols, aiming for biomedical applications. The synthesis is based on a process in which polyethylene glycol (PEG400) is used as phase-transfer catalyst between fullerene/benzene and aqueous NaOH solutions. The polyhydroxylation of the fullerenes occurs in the NaOH solution under a continuous flow of O2 to enhance the reaction yield. The resulting compound was characterized with infrared spectroscopy, nuclear magnetic resonance, thermo-gravimetric analysis and optical absorption. The formation of C60(OH)18-20 in high yields was confirmed.

Structural and photophysical properties of peptide micro/nanotubes functionalized with hypericin.

Hypericin is a photosensitizer with promising applications in photodynamic therapy (PDT) for cancer and infectious diseases treatments. Herein, we present a basic research study of L-diphenylalanine micro/nanotubes (FF-NTs) functionalized with hypericin. The system has special properties according to the hypericin concentration, with direct consequences on both morphological and photophysical behaviors. A clear dependence between the size of the tubes and the concentration of hypericin is revealed. The generation of reactive oxygen species (ROS) is found to be improved by ∼57% in the presence of FF-NTs, as indirectly measured from the absorbance profile of 1,3-diphenylisobenzofuran (DPBF). In addition, when hypericin appears conjugated with FF-NTs, the characteristic fluorescence lifetime is significantly boosted, demonstrating the role of FF-NTs to enhance the photophysical properties and stabilizing the fluorophore in excited states. Electron paramagnetic resonance allows the proposition of a mechanism for the generation of ROS. Molecular dynamics simulations bring new insights into the interaction between hypericin and peptide assemblies, suggesting the spatial organization of the fluorophore onto the surface of the supramolecular structures as a key element to improve the photophysical properties reported here.

Gene expression and biochemical responses in brain of zebrafish Danio rerio exposed to organic nanomaterials: carbon nanotubes (SWCNT) and fullerenol (C60(OH)18-22(OK4)).

Nanomaterials (NM) industry had grown in the last decade, although there are few studies concerning its potential toxicity effects on aquatic organisms. In this study the freshwater zebrafish (Danio rerio) was exposed to two kinds of carbon NM, single-wall carbon nanotubes (SWCNT) and fullerenol [C60(OH)18-22(OK4)] to analyze oxidative stress responses on fish brain. Adult zebrafish (mean mass: 0.52±0.01g) were submitted to intraperitoneal injections of SWCNT suspension and fullerenol solution (30mg/kg of fish), receiving one or two doses with a time interval of 24h. Results showed that total antioxidant capacity was lowered in brains of fish exposed 24h to fullerenol when compared to those from SWCNT treatment (p<0.05). After 48h, fullerenol induced higher expression of both catalytic and regulatory subunits of enzyme glutamate cysteine ligase when compared to control group (p<0.05), indicating an antioxidant behavior. In vitro assays showed a dual effect of SWCNT, since a pro-oxidant behavior was observed at low concentrations (0.1 and 1.0mg/L) and an antioxidant one at the highest concentration (10.0mg/L). Few biological responses were altered by this NM: decrease in total antioxidant capacity and induction of the expression of the transcription factor Nrf2 when compared to control group.

In vitro exposure to fullerene C60 influences redox state and lipid peroxidation in brain and gills from Cyprinus carpio (Cyprinidae)

Studies concerning the impact of nanomaterials, especially fullerene (C(60) ), in fresh water environments and their effects on the physiology of aquatic organisms are still scarce and conflicting. We aimed to assess in vitro effects of fullerene in brain and gill homogenates of carp Cyprinus carpio, evaluating redox parameters. A fullerene suspension was prepared by continued stirring under fluorescent light during two months. The suspension concentration was measured by total carbon content and ultraviolet-visible spectroscopy nephelometry. Characterization of C(60) aggregates was performed with an enhanced dark-field microscopy system and transmission electronic microscopy. Organ homogenates were exposed during 1, 2, and 4 h under fluorescent light. Redox parameters evaluated were reduced glutathione and oxidized glutathione, cysteine and cystine, total antioxidant capacity; activity of the antioxidant enzymes glutathione S-transferase and glutathione reductase (GR), and lipid peroxidation (TBARS assay). Fullerene induced a significant increase (p < 0.05) in lipid peroxidation after 2 h in both organs and reduced GR activity after 1 h (gills) and 4 h (brain) and antioxidant capacity after 4 h (brain). Levels of oxidized glutathione increased in the brain at 1 h and decreased at 2 h as well. Given these results, it can be concluded that C(60) can induce redox disruption via thiol/disulfide pathway, leading to oxidative damage (higher TBARS values) and loss of antioxidant competence.

Quantification of fullerene nanoparticles suspensions in water based on optical scattering.

Fullerenes, and in particular the C60, have been intensively investigated in the last decades mainly because of their vast range of potential applications in biomedicine and materials science. These molecules are inherently hydrophobic, tending thus to form clusters and aggregates in polar solvents resulting in colloidal suspensions. In this work was developed a quantification method for C60 nanoparticles (nano-C60) in colloidal aqueous suspensions based on optical light scattering (nephelometry). This method can be done in a conventional spectrofluorimeter either on the excitation wavelengths or on the second-order satellite lines that arise from the diffraction gratings. The detection limit of the proposed method was about (0.0090+/-0.0008) mg L(-1), in a linear concentration range from 0.007 to 0.360 mg L(-1). A comparison of this scattering technique with spectrophotometry based on molecular absorption shows that for the former, even at the second-order lines, the threshold concentrations detected are about 20 times lower that the latter.

The reduction of oxidative stress by nanocomposite Fullerol decreases mucositis severity and reverts leukopenia induced by Irinotecan.

Irinotecan is a useful chemotherapeutic agent for the treatment of several solid tumors. However, this therapy is associated with side effects, including leukopenia and mucositis. Reactive oxygen species (ROS) activate inflammatory pathways and contribute to Irinotecan-induced mucositis. Fullerol is a nanocomposite with anti-oxidant properties that may reduce tissue damage after inflammatory stimuli. In this paper, the effects of Fullerol and mechanisms of protection were investigated in a model of Irinotecan-induced mucositis. Mucositis was induced by an injection of Irinotecan per 4 days in C57BL/6. Fullerol or a vehicle was injected every 12h. On day 7, the intestines were removed to evaluate histological changes, leukocyte influx, and the production of cytokines and ROS. Irinotecan therapy resulted in weight loss, an increased clinical score and intestinal injury. Treatment with Fullerol attenuated weight loss, decreased clinical score and intestinal damage. Irinotecan also induced increased ROS production in enterocytes, oxidative stress, IL-1β production, neutrophil and eosinophil influx in the ileum. Fullerol treatment decreased production of ROS in the enterocytes, oxidative stress, IL-1β production, neutrophil and eosinophil influx in the ileum. Irinotecan therapy also induced leukopenia in an ROS-dependent manner because leukopenia reverted in WT mice treated with Fullerol or Apocynin or in Gp91phox(-/-) mice. Mice treated with Irinotecan presented less melanoma tumor growth compared to the control group. Fullerol does not interfere in the anti-tumor action of Irinotecan. Fullerol has a great pharmacology potential to decreases the severity of mucositis and of leukopenia during chemotherapy treatment.

Spectroscopic characterization of transition metal impurities in natural montebrasite/amblygonite

Abstract Natural single-crystal specimens of the montebrasite/amblygonite series from Brazil, with general formula LiAlPO4(F,OH), were investigated by electron microprobe, Raman spectroscopy, X-ray diffraction, and infrared absorption. Since little is known about impurities and their local symmetries, electron paramagnetic resonance (EPR) was applied. Six different paramagnetic impurities and radiation defects were detected by EPR. Three of them, all substituting for Al3+ ions, namely, iron (Fe3+), vanadium (V4+), and niobium (Nb4+) impurities were characterized in this work. The Fe3+ (3d5)-related EPR spectra and angular dependencies show occupation of low-symmetry sites that are revealed in the high asymmetry parameter of the electronic fine structure, E/D = 0.27. Vanadium and niobium impurities are identified through their typical strong hyperfine interactions. Both form interesting examples for which the properties of 3d1 ion (V4+) and 4d1 ion (Nb4+) in the same host matrix can be compared. It is shown that both ions form complex defects of type VO2+ (vanadyl) and NbO2+ (niobyl), showing superhyperfine interaction with two equivalent hydrogen ions and not to fluorine. The EPR rotation patterns are analyzed in detail for three mutually perpendicular crystal planes. Spin Hamiltonian parameters are calculated and discussed.

Hybrid systems based on gold nanostructures and porphyrins as promising photosensitizers for photodynamic therapy

Gold nanostructures of two different shapes (spheres and rods) were synthesized to form a colloidal hybrid system with 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)porphyrin tosylate salt (H2TM4PyP(OTs)4) (POR) for applications in photodynamic therapy (PDT) using light in the visible spectral range. Electron paramagnetic resonance (EPR) experiments in combination with spin trapping were used for the detection of reactive oxygen species (ROS) and evaluation of the efficiency of these novel hybrid systems as photosensitizers. It is shown that the hybrid system consisting of gold nanorods (AuNR) and porphyrin (POR) is by far more efficient than its isolated components. This enhanced efficiency is explained by a synergetic effect between the AuNR and the porphyrin, wherein a rapid energy transfer from the former to the latter produces a large amount of singlet oxygen followed by its conversion into hydroxyl radicals. The mechanism was investigated using different spin traps and different ROS inhibitors. On the other hand, spherical gold nanoparticles (AuNP) do not show this synergetic effect. The synergetic effect for gold nanorods/POR hybrid is attributed to a larger field enhancement close to the gold nanorod surface in addition to the electrostatic attraction between the components of the hybrid system.

New material for low-dose brachytherapy seeds: Xe-doped amorphous carbon films with post-growth neutron activated 125I

We report a novel material for use in (125)I brachytherapy that consists of amorphous carbon films grown by ion-beam-assisted deposition and doped with Xe (5 at%) by implantation. Samples of these films grown on Si substrates were irradiated with neutrons in a TRIGA-I nuclear reactor for the production (125)Xe, and latter characterized by gamma spectroscopy. The results indicate that the (124)Xe was efficiently converted into (125)Xe, the precursor of (125)I, and support the activity calculations for a model brachytherapy seed.