Water-soluble C60 Fullerene Research Articles

Analysis of Biomechanical Parameters of Muscle Soleus Contraction and Blood Biochemical Parameters in Rat with Chronic Glyphosate Intoxication and Therapeutic Use of C60 Fullerene.

The widespread use of glyphosate as a herbicide in agriculture can lead to the presence of its residues and metabolites in food for human consumption and thus pose a threat to human health. It has been found that glyphosate reduces energy metabolism in the brain, its amount increases in white muscle fibers. At the same time, the effect of chronic use of glyphosate on the dynamic properties of skeletal muscles remains practically unexplored. The selected biomechanical parameters (the integrated power of muscle contraction, the time of reaching the muscle contraction force its maximum value and the reduction of the force response by 50% and 25% of the initial values during stimulation) of muscle soleus contraction in rats, as well as blood biochemical parameters (the levels of creatinine, creatine phosphokinase, lactate, lactate dehydrogenase, thiobarbituric acid reactive substances, hydrogen peroxide, reduced glutathione and catalase) were analyzed after chronic glyphosate intoxication (oral administration at a dose of 10 μg/kg of animal weight) for 30 days. Water-soluble C60 fullerene, as a poweful antioxidant, was used as a therapeutic nanoagent throughout the entire period of intoxication with the above herbicide (oral administration at doses of 0.5 or 1 mg/kg). The data obtained show that the introduction of C60 fullerene at a dose of 0.5 mg/kg reduces the degree of pathological changes by 40–45%. Increasing the dose of C60 fullerene to 1 mg/kg increases the therapeutic effect by 55–65%, normalizing the studied biomechanical and biochemical parameters. Thus, C60 fullerenes can be effective nanotherapeutics in the treatment of glyphosate-based herbicide poisoning.

Anticoronavirus Activity of Water-Soluble Pristine C60 Fullerenes: In Vitro and In Silico Screenings.

The emergence of a new member of the Coronaviridae family, which caused the 2020 pandemic, requires detailed research on the evolution of coronaviruses, their structure and properties, and interaction with cells. Modern nanobiotechnologies can address the many clinical challenges posed by the COVID-19 pandemic. In particular, they offer new therapeutic approaches using biocompatible nanostructures with "specific" antiviral activity. Therefore, the nanosized spherical-like molecule (0.72 nm in diameter) composed of 60 carbon atoms, C60 fullerene, is of interest in terms of fighting coronaviruses due to its high biological activity. In here, we aim to evaluate the effectiveness of anticoronavirus action of water-soluble pristine C60 fullerene in the model and in vitro systems. As a model, apathogenic for human coronavirus, we used transmissible gastroenteritis virus of swine (TGEV), which we adapted to the BHK-21 cell culture (kidney cells of a newborn Syrian hamster). The shape and size of the particles present in C60 fullerene aqueous colloidal solution (C60FAS) of given concentration, as well as C60FAS stability (value of zeta potential) were studied using microscopic (STM, scanning tunneling microscopy, and AFM, atomic force microscopy) and spectroscopic (DLS, dynamic light scattering) methods. The cytopathic effect of TGEV was determined with the help of a Leica DM 750 microscope and the degree of monolayer changes in cells was assessed. The microscopy of the viral suspension was performed using a high resolution transmission electron microscope (HRTEM; JEM-1230, Japan). Finally, the search for and design of optimal possible complexes between C60 fullerene and target proteins in the structure of SARS-CoV-2 coronavirus, evaluation of their stability in the simulated cellular environment were performed using molecular dynamics and docking methods. It was found that the maximum allowable cytotoxic concentration of C60 fullerene is 37.5±3.0μg/ml. The investigated C60FAS reduces the titer of coronavirus infectious activity by the value of 2.00±0.08TCID50/ml. It was shown that C60 fullerene interacts directly with SARS-CoV-2 proteins, such as RdRp (RNA-dependent RNA polymerase) and 3CLpro (3-chymotrypsin-like protease), which is critical for the life cycle of the coronavirus and, thus, inhibits its functional activity. In both cases, C60 fullerene fills the binding pocket and gets stuck there through stacking and steric interactions. Pioneer in vitro study to identify the anticoronavirus activity of water-soluble pristine C60 fullerenes indicates that they are highly promising for further preclinical studies, since a significant inhibition of the infectious activity of swine coronavirus of transmissible gastroenteritis in BHK-21 cell culture was found. According to molecular modeling results, it was shown that C60 fullerene can create the stable complexes with 3CLpro and RdRp proteins of SARS-CoV-2 coronavirus and, thus, suppress its functional activity.

Physicochemical properties, biological activity and biocompatibility of water-soluble C60-Hyp adduct

Amino acid adducts of light fullerenes have a potential of application in a variety of fields of biomedicine, that is reactive oxygen species scavenging activity, anticancer activity, viruses and bacteria inactivation etc. In this work, the water-soluble C60 fullerene derivative with l-hydroxyproline (C60(C5H9NO3)2, C60-Hyp) was studied. Extensive biomedical investigation of this compound, namely, antiradical activity in the reaction with stable diphenylpicrylhydrazyl radical, the binding to human serum albumin, photodynamic properties, cytotoxicity in glioblastoma A172 and lung carcinoma A549 cell lines, erythrocytes haemolysis, platelet aggregation, genotoxicity on human peripheral blood mononuclear cells was conducted. Moreover, the dynamic and structural characteristics of C60-Hyp–H2O binary system were obtained using molecular dynamic (MD) method, and size distribution along with ζ-potentials of C60-Hyp associates was measured.

Water-Soluble Pristine C60 Fullerene Inhibits Liver Alterations Associated with Hepatocellular Carcinoma in Rat

Excessive production of reactive oxygen species is the main cause of hepatocellular carcinoma (HCC) initiation and progression. Water-soluble pristine C60 fullerene is a powerful and non-toxic antioxidant, therefore, its effect under rat HCC model and its possible mechanisms were aimed to be discovered. Studies on HepG2 cells (human HCC) demonstrated C60 fullerene ability to inhibit cell growth (IC50 = 108.2 μmol), to induce apoptosis, to downregulate glucose-6-phosphate dehydrogenase, to upregulate vimentin and p53 expression and to alter HepG2 redox state. If applied to animals experienced HCC in dose of 0.25 mg/kg per day starting at liver cirrhosis stage, C60 fullerene improved post-treatment survival similar to reference 5-fluorouracil (31 and 30 compared to 17 weeks) and inhibited metastasis unlike the latter. Furthermore, C60 fullerene substantially attenuated liver injury and fibrosis, decreased liver enzymes, and normalized bilirubin and redox markers (elevated by 1.7–7.7 times under HCC). Thus, C60 fullerene ability to inhibit HepG2 cell growth and HCC development and metastasis and to improve animal survival was concluded. C60 fullerene cytostatic action might be realized through apoptosis induction and glucose-6-phosphate dehydrogenase downregulation in addition to its antioxidant activity.

Singlet oxygen generation by sonication using a water-soluble fullerene (C60) complex: a potential application for sonodynamic therapy

The generation of singlet oxygen (1O2) by sonicating a water-soluble complex of fullerene/poly(2-methacryloyloxyethyl phosphorylcholine) (C60/PMPC) was investigated for sonodynamic therapy (SDT). The effect of sonication time and the solubilized concentration of water-soluble fullerene C60 on the amount of generated 1O2 was studied. Singlet oxygen sensor green was used to confirm the generation of 1O2. 1O2 was generated from C60/PMPC under ultrasonic irradiation, and the amount of 1O2 increased with increasing solubilized C60 concentration and irradiation time. In cell viability tests, after 3 h of incubation, the cell viabilities were 10 and 84% in the presence of C60/PMPC with and without sonication, respectively. This result suggests that C60/PMPC shows a sonodynamic effect, which demonstrates its potential application in SDT. The generation of singlet oxygen (1O2) by sonicating a water-soluble complex of fullerene/poly(2-methacryloyloxyethyl phosphorylcholine) (C60/PMPC) was investigated for sonodynamic therapy (SDT). 1O2 was generated from C60/PMPC under ultrasonic irradiation, and the amount of 1O2 increased with increasing solubilized C60 concentration and irradiation time. In cell viability tests, after 3 h of incubation, the cell viabilities were 10% and 84% in the presence of C60/PMPC with and without sonication, respectively. This result suggests that C60/PMPC shows a sonodynamic effect, which demonstrates its potential application in SDT.

The effect of fullerenol C60 on skeletal muscle after lower limb ischemia reperfusion injury in streptozotocin-induced diabetic rats

Aim: Fullerenol, a water-soluble C60-fullerene, has been demonstrated to scavenge free radicals in vitro and in vivo. The aim of the study was to investigate the effects of fullerenol C60 on lower skeletal muscles in a rat model of ischemia/reperfusion (I/R).Methods: After approval of the ethics committee, 30 Wistar Albino rat were divided into 5 groups with six animals per each as follows: Control (C), diabetes (D), diabetes+fullerenol C60 (DF), diabetes+I/R (group DIR) and diabetes I/R+fullerenol C60 (DIR-F) groups. Streptozotocin was administered to the rats to induce diabetes at a dose of 55 mg/kg. Four weeks after the onset of diabetes, rats were subjected to 2 hours of ischemia and 2 hours of reperfusion. At the end of the reperfusion period, skeletal muscle samples were taken from the lower extremity in all groups for histopathological and immunohistopathological examinations. Results: Myositis and endothelial caspase 3 enzyme activities were high in all groups, particularly DIR. Compared to C, DF and DIR-F groups, inflammation and myositis were significantly higher in the DIR group (P=0.001, P=0.006, P=0.001, respectively, and P=0.001, P=0.022, P=0.001, respectively). Vascular dilatation and congestion were significantly more prominent in all groups compared to the control group (P=0.001 for all).Conclusion: Our results confirm that fullerenol C60 has protective effects against skeletal muscle damage resulting from I/R in diabetic rats. Future studies conducted to evaluate these effects may help illuminate the action mechanism of fullerenol C60 and pathophysiology underlying the tissue damage related to I/R injury.

Evaluation of the Biocompatibility of Water-Soluble Pristine С60 Fullerenes in Rabbit

C60 fullerenes have proved their therapeutic effects and efficacy by the results of countless experiments. For further usage of these nanoparticles, the systematic toxicological investigations are required. Blood compatibility should be studied for C60 fullerenes due to the potential blood contact. Currently, available data is not systematic and has not provided insights into possible side effects of C60 fullerenes on blood components. In this study, water-soluble pristine C60 fullerenes were tested in vitro to assess their biocompatibility in rabbit. The blood compatibility has been evaluated looking at the impact of C60 fullerenes on erythrocyte integrity, platelet aggregation, and some blood factors involved in coagulation. Our results revealed that C60 fullerenes cannot elicit hemolysis at studied concentrations and did not show any effect on coagulation process. C60 fullerenes in concentration-dependent manner increased ADP-dependent platelet aggregation and changed the key kinetic parameters of these processes. C60 fullerenes inhibited thrombin amidolytic activity but did not affect the activities of other studied coagulation factors. The prothrombotic property of C60 fullerenes could be the potential risk factor that leads to enhancement of vascular thrombosis. The ability of fullerene to inhibit thrombin activity is important for the pharmacological use of these carbon nanoparticles as anticoagulant agents.

Water-Soluble Anionic C60-Fullerene Derivatives as Antidotes for HG(II) Ions in Tests on Escherichia Coli Cells

Three water-soluble C60-fullerene derivatives functionalized by anionic adducts of various chemical compositions and structures were prepared in highly specific reactions of chlorofullerene C60Cl6. Hybrid metal-organic complexes formed and aggregated in several instances in aqueous solution in the presence of stoichiometric (1:1 by moles) amounts of synthesized C60-fullerene derivatives and Hg(II) ions. The bioavailability of Hg(II) ions for Escherichia coli cells was reduced by such a reaction and manifested as decreased toxicity for strain E. coli K12 TG1 pF1 or weakened protective reactions of strain E. coli K12 MG1655 pMerA'::lux. The most pronounced protective effect was recorded for the C60-fullerene derivative functionalized by S-containing adducts of general formula –S(CH2)10CO2K with antidote activity comparable with that of the pharmacopoeial preparation Unithiol.

Dynamics of Contraction of the musculus soleus of Rats in Chronic Alcoholization and Therapeutic Action of Water-Soluble C60 Fullerenes

Dynamics of Contraction of the musculus soleus of Rats in Chronic Alcoholization and Therapeutic Action of Water-Soluble C60 Fullerenes

Reconciling the controversial data on the effects of C60 fullerene at the organismal and molecular levels using as a model Drosophila melanogaster

Fullerenes have attracted attention since their discovery as structural units of complex carbon nanostructures capable of transporting drugs and macromolecules. As such artificial nanomaterials are applied in biology and medicine, they are routinely scrutinized for their effects on living organisms. The results of such studies range from direct destabilizing effects on DNA molecules to amelioration of the toxic effects of known genotoxic agents. We tested the effect of buckminsterfullerene (C60) on Drosophila melanogaster at DNA, tissue and organism levels. The water-soluble pristine C60 fullerene at the concentration of 20 μg/ml and 40 μg/ml leads to the activation of the mus209 gene in D. melanogaster larvae salivary glands, which can indicate higher levels of DNA damage. However, the absence of effects at the cell and organismal level could be explained by the activation of repair systems or by active elimination of damaged cells.

C60 fullerenes selectively inhibit BKCa but not Kv channels in pulmonary artery smooth muscle cells

Possessing unique physical and chemical properties, C60 fullerenes are arising as a potential nanotechnological tool that can strongly affect various biological processes. Recent molecular modeling studies have shown that C60 fullerenes can interact with ion channels, but there is lack of data about possible effects of C60 molecule on ion channels expressed in smooth muscle cells (SMC). Here we show both computationally and experimentally that water-soluble pristine C60 fullerene strongly inhibits the large conductance Ca2+-dependent K+ (BKCa), but not voltage-gated K+ (Kv) channels in pulmonary artery SMC. Both molecular docking simulations and analysis of single channel activity indicate that C60 fullerene blocks BKCa channel pore in its open state. In functional tests, C60 fullerene enhanced phenylephrine-induced contraction of pulmonary artery rings by about 25% and reduced endothelium-dependent acetylcholine-induced relaxation by up to 40%. These findings suggest a novel strategy for biomedical application of water-soluble pristine C60 fullerene in vascular dysfunction.

Antioxidant Properties of Fullerene Derivatives Depend on Their Chemical Structure: A Study of Two Fullerene Derivatives on HELFs.

Oxidative stress is a major issue in a wide number of pathologies (neurodegenerative, cardiovascular, immune diseases, and cancer). Because of this, the search for new antioxidants is an important issue. One of the potential antioxidants that has been enthusiastically discussed in the past twenty years is fullerene and its derivatives. Although in aqueous solutions fullerene derivatives have shown to be antioxidants, their properties in this regard within the cells are controversially discussed. We have studied two different water-soluble fullerene C60 and C70 derivatives on human embryonic lung fibroblasts at a wide range of concentrations. Both of them cause a decrease in cellular ROS at short times of incubation (1 hour). Their prolonged action, however, is fundamentally different: derivative GI-761 causes secondary oxidative stress whereas derivative VI-419-P3K keeps ROS levels under control values. To gain a better understanding of this effect, we assessed factors that could play a role in the response of cells to fullerene derivatives. Increased ROS production occurred due to NOX4 upregulation by GI-761. Derivative VI-419-P3K activated the transcription of antioxidant master regulator NRF2 and caused its translocation to the nucleus. This data suggests that the antioxidant effect of fullerene derivatives depends on their chemical structure.

Water-soluble pristine C60 fullerene attenuates acetaminophen-induced liver injury.

Introduction: Oxidative stress has been suggested as the main trigger and pathological mechanism of toxic liver injury. Effects of powerful free radical scavenger С60 fullerene on rat liver injury and liver cells (HepG2 line) were aimed to be discovered. Methods: Acute liver injury (ALI) was simulated by single acetaminophen (APAP, 1000 mg/kg) administration, on a chronic CLI, by 4 weekly APAP administrations. Pristine C60 fullerene aqueous colloid solution (C60FAS; initial concentration 0.15 mg/mL) was administered per os or intraperitoneally at a dose of 0.5 mg/kg (ALI) or 0.25 mg/kg (CLI) daily for 2 or 28 days, respectively, after first APAP dose. Animals were sacrificed at 24th hour after the last dose. Biochemical markers of blood serum and liver autopsies were analyzed. EGFR expression in HepG2 cells after 48-hour incubation with C60FAS was assessed. Results: Increase of serum conjugated and unconjugated bilirubin (up to 1.4-3.7 times), ALT (by 31-37%), and AST (by 18%) in non-treated ALI and CLI rats were observed, suggesting the hepatitis (confirmed by histological analysis). Liver morphological state (ALI, CLI), ALT (ALI and CLI), bilirubin (CLI), α-amylase, and creatinine (ALI) were normalized with C60FAS administration in both ways, which may indicate its protective impact on liver. However, unconjugated bilirubin sharply increased in ALI animals receiving C60FAS (up to 12 times compared to control), suggesting the augmentation of bilirubin metabolism. Furthermore, C60FAS inhibited EGFR expression in HepG2 cells in a dose-dependent manner. Conclusion: C60FAS could partially correct acute and chronic toxic liver injury, however, it could not normalize bilirubin metabolism after acute exposure.

Impact of Water-Soluble C60 Fullerenes on the Mechanokinetic Features of Formation of a Smooth Tetanic Contraction of Ischemic Skeletal Muscle of Rats

Impact of Water-Soluble C60 Fullerenes on the Mechanokinetic Features of Formation of a Smooth Tetanic Contraction of Ischemic Skeletal Muscle of Rats

Fullerol C60(OH)24 nanoparticles modulate aflatoxin B1 biosynthesis in Aspergillus flavus

The water soluble fullerene C60 daughter product - fullerols C60(OH)24 (FNP) possesses a great potential of modifying secondary metabolites biosynthesis. In order to clarify the extent of interaction, the impact of FNP (10, 100 and 1000 ng mL−1) on aflatoxin production and the available precursors of biosynthesis pathway from Aspergillus flavus NRRL 3251 was determined, in both the mycelia and yeast extract sucrose (YES) medium, during a 168-hour growth period at 29 °C in the dark. The FNP of 8 nm in diameter, and with a zeta potential of −33 mV affected mycelial growth at 1000 ng mL−1 while conidia production was slightly affected at 10 ng mL−1. The FNP effect on aflatoxin and it biosynthetic precursors was concentration dependent and alteration of the sterigmatocystin (ST) export from the cell was observed. Most of the monitored aflatoxin precursors, except norsolorinic acid, were detected in both mycelia and YES medium. However, observed precursor concentrations were much higher in mycelia, with exception of ST. The study shows the loss of FNP antioxidative effect after 120 hours of growth, and strong concentration dependent aflatoxigenic effect after that time. Thus, this data is relevant to guide future considerations on FNP-fungal interactions in the environments and on risk assessment.

A Novel Nanoconjugate of Landomycin A with C60 Fullerene for Cancer Targeted Therapy: In Vitro Studies.

Landomycins are a subgroup of angucycline antibiotics that are produced by Streptomyces bacteria and possess strong antineoplastic potential. Literature data suggest that enhancement of the therapeutic activity of this drug may be achieved by means of creating specific drug delivery systems. Here we propose to adopt C60 fullerene as flexible and stable nanocarrier for landomycin delivery into tumor cells. The methods of molecular modelling, dynamic light scattering and Fourier transform infrared spectroscopy were used to study the assembly of C60 fullerene and the anticancer drug Landomycin A (LA) in aqueous solution. Cytotoxic activity of this nanocomplex was studied in vitro towards two cancer cell lines in comparison to human mesenchymal stem cells (hMSCs) using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) test and a live/dead assay. The morphology of the cells incubated with fullerene-drug nanoparticles and their uptake into target cells were studied by scanning electron microscopy and fluorescence light microscopy. The viability of primary cells (hMSCs, as a model for healthy cells) and cancer cell lines (human osteosarcoma cells, MG-63, and mouse mammary cells, 4T1, as models for cancer cells) was studied after incubation with water-soluble C60 fullerenes, LA and the mixture C60 + LA. The C60 + LA nanocomplex in contrast to LA alone showed higher toxicity towards cancer cells and lower toxicity towards normal cells, whereas the water-soluble C60 fullerenes at the same concentration were not toxic for the cells. The obtained physico-chemical data indicate a complexation between the two compounds, leading to the formation of a C60 + LA nanocomposite. It was concluded that immobilization of LA on C60 fullerene enhances selectivity of action of this anticancer drug in vitro, indicating on possibility of further preclinical studies of novel C60 + LA nanocomposites on animal tumor models.

Lysine-Based C60-Fullerene Nanoconjugates for Monomethyl Fumarate Delivery: A Novel Nanomedicine for Brain Cancer Cells.

In the present study, water-soluble lysine-based C60-fullerene nanoconjugates (CF-LYS-TEG-MMF) were synthesized using a biodegradable linker for the better delivery of monomethyl fumarate (MMF) employing Prato reaction. CF-LYS-TEG-MMF resulted in enhanced cytotoxicity on neuroblastoma cells, meanwhile found to be substantially biocompatible to erythrocytes. The designed nanoconjugate exhibited a pH-based drug release pattern, minimizing the leaching of drug at plasma pH. However, the carrier offered maximum drug release at cancer cell pH, indicating huge promise in internalization of drug molecules at the site of target. The pharmacokinetics of MMF in rodents was significantly improved in terms of enhanced bioavailable drug fraction in the central compartment, reduced drug clearance, elevated plasma concentrations and prolonged biological residence of drug. Enhanced in vitro efficacy in SH-SY5Y neuroblastoma cells, improved erythrocyte compatibility, high drug loading, and conducive pharmacokinetic profile by CF-LYS-TEG-MMF offers a huge promise in brain drug delivery, dose reduction, and dosage-regimen alteration for the management of brain tumors employing MMF.

Water-soluble C60 fullerenes reduce manifestations of acute cholangitis in rats

Sclerosing cholangitis is the liver disease of uncertain etiology, extremely unfavorable prognosis and lack of effective medication therapy. Therefore, the effect of water-soluble biocompatible C60 fullerenes (C60FAS) on the liver functional state on rat acute-cholangitis model was aimed to be discovered. Acute cholangitis was simulated by single α-naphthyl isothiocyanate (ANIT, 100 mg/kg) per os administration; C60FAS (0.5 mg/kg) was administered either per os or intraperitoneally in 24 and 48 h after ANIT ingestion, and in 72 h the animals were sacrificed. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), the total and direct bilirubin, creatinine and urea in the blood serum were determined, and the liver morphological state was assessed. In animals experienced ANIT-induced acute cholangitis, the total and direct bilirubin, creatinine, ALT, AST, ALP and LDH 1.5–4-fold increase were observed, indicating cytolysis of hepatocytes, cholestasis, and renal dysfunction. The features of periductal fibrosis, biliary epithelium atrophy, and portal–portal linking septa formation were detected, confirming the sclerosing cholangitis development. C60FAS promoted to the normalization of direct and total bilirubin levels, the ALT activity and diminution of fibrotic features. In addition, C60FAS intraperitoneal administration also normalized the ALP activity, indicating the attenuation of disease symptoms. However, the AST activity and creatinine level remained unchanged, and the LDH activity even increased, manifesting the partial persistence of cholestasis and renal dysfunction. Thus, the therapeutic application of C60FAS promotes a partial protection of liver against cholangitis.

Effect of C60 fullerene nanoparticles on the diet-induced obesity in rats

Obesity is a growing global health problem. Since increased oxidative stress is one of the key pathological mechanisms underpinning overweight and strongly correlates with progression of obesity-related complications we hypothesized that C60 fullerene nanoparticles, due to their strong antioxidant capacity, could be the promising therapeutic agent in the treatment of this disease. Here we investigated whether the C60 fullerenes can alleviate diet-induced obesity (DIO) and metabolic impairments associated with it. To determine the effect of C60 fullerenes on some nutritional and metabolic parameters, rats were fed either a normal diet (6.7% fat, 15.27 kJ·g-1) or a high-fat diet (38.8% fat, 28.71 kJ·g-1) for 70 days and were simultaneously treated per os with pristine C60 fullerene aqueous solution (C60FAS; 0.3 mg·kg-1 every other day) since the 28th day from the start of the experiment. Rats fed with high fat diet had significantly increased body mass index (BMI), levels of insulin, glucose, glycosilated hemoglobin (HbA1c) and serum pro-inflammatory cytokines compared with control rats fed with low-fat chow. C60 fullerenes normalized the metabolic parameters and partially reduced BMI in DIO animals. Pro-inflammatory cytokines (IL-1b, IL-12, INFγ) were also decreased in serum of DIO rats treated with C60 fullerenes while anti-inflammatory cytokines (IL-4, IL-10) were at the control levels. High fat diet caused the increased level of oxidative stress products, and this was accompanied by decreased activity both the superoxide dismutase and catalase, whereas the administration of C60 fullerenes markedly decreased level of oxidative stress and enhanced antioxidant enzyme activities. These data indicate that water-soluble pristine C60 fullerenes reduce chronic inflammation, restore glucose homeostasis as well as positively affects on prooxidant-antioxidant homeostasis. C60 fullerenes could be represented as a promising therapeutic agent in the treatment of obesity and its related complications.

C60 fullerenes disrupt cellular signalling leading to TRPC4 and TRPC6 channels opening by the activation of muscarinic receptors and G-proteins in small intestinal smooth muscles

The effect of water-soluble pristine C60 fullerene nanoparticles (C60NPs) on receptor-operated cation channels formed by TRPC4/C6 proteins in ileal smooth muscle cells was investigated for the first time. Activation of these channels subsequent to acetylcholine binding to the expressed in these cells M2 and M3 muscarinic receptors represents the key event in the parasympathetic control of gastrointestinal smooth muscle motility and cholinergic excitation-contraction coupling. Experiments were performed on single collagenase-dispersed mouse ileal myocytes using patch-clamp techniques with symmetrical 125mM Cs+ solutions and [Ca2+]i ‘clamped’ at 100nM in order to isolate the muscarinic cation current (mICAT). The current was induced by intracellular infusion of 200μM GTPγS, which activates G-proteins directly, i.e. bypassing the muscarinic receptors. C60NPs applied at 10−6M at peak response to activation of G-proteins caused mICAT inhibition by 47.0±3.5% (n=9). The inhibition developed rather slowly, with the time constant of 119±16s, was voltage-independent and irreversible. Thus, C60NPs are unlikely to cause any direct block of TRPC4/C6 channels; rather, they may accumulate in the membrane and disrupt G-protein signalling leading to mICAT generation. C60NPs may represent a novel class of biocompatible molecules for the treatment of disorders associated with enhanced gastrointestinal motility.