Bioactivity of cerium dioxide nanoparticles as a function of size and surface features.

Nano-dispersed cerium dioxide is promising for use in medicine due to its unique physicochemical properties, including low toxicity, the safety of in vivo usage, active participation in different redox processes occurring in living cells, and its regenerative potential, manifested in the ability of CeO2 to participate repeatedly in redox reactions. In this work, we examined the biological activity of cerium dioxide nanoparticles (CeO2 NPs) synthesized by precipitation in mixed water/alcohol solutions at a constant pH of 9. This synthesis method allowed controlling the size and Ce3+/Ce4+ proportion on the surface of NPs, changing the synthesis conditions and obtaining highly stable suspensions of "naked" CeO2 NPs. Changes in the surface properties upon contact of CeO2 NPs with protein-rich media, e.g., bovine serum albumin and DMEM cell culture medium supplemented with 10% fetal bovine serum, the characteristics of nanoparticle uptake by mouse aortic endothelial cells and the antioxidant activity of the nanoparticles of different sizes were investigated by various state-of-the-art analytical methods.

Myeloprotection with activated carbon in doxorubicin-treated rats.

Chemotherapy can often cause a variety of side effects including bone marrow (BM) suppression, termed as myelosuppression. Accordingly, facile and effective management of chemotherapy-induced myelosuppression is currently a pivotal task for experimental pathologists and oncologists. Here, we chose to use activated carbon (AC) with an extensive surface area for studying its possible protective effectiveness with respect to BM in doxorubicin (DOX)-treated rats. Spherical AC with an extended surface area up to 4490 m2/g was prepared for per os (p/o) delivery, whereas for intraperitoneal (i/p) delivery we used the powdered form of AC that was derived from the aforementioned spherical AC. During the monthly treatment of animals with AC and DOX these two components were delivered alternately (not in the same day). After treatment, BM cells were isolated from femurs of sacrificed animals, stained with acridine orange (AO) and analyzed by flow cytometry. Regardless of the route of AC delivery (p/o or i/p), apparent myeloprotection with a possible regenerative effect was observed in animals that received DOX, as evidenced by recovery of the populations of total nucleated cells (TNC) and polychromatic (immature) erythrocytes accompanied by a considerable reduction of the number of apoptotic/dead cells among TNC (≤2.0%). Moreover, as a result of AC administrations, there was a significant increase of AO green and far-red fluorescence intensities in the population of TNC, which is suggestive of the ongoing quantitative and conformational changes in DNA and RNA associated with cell recovery and proliferation. Thus, AC preparations under the present experimental conditions can effectively tackle DOX-induced myelosuppression via mechanisms not necessarily associated with adsorptive detoxification.

Cerium dioxide nanoparticles synthesized via precipitation at constant pH: Synthesis, physical-chemical and antioxidant properties.

Cerium oxide nanoparticles (CeO2 NPs) are well known for their application in various fields of industry, as well as in biology and medicine. Knowledge of synthesis schemes, physicochemical and morphological features of nanoscale CeO2 is important for assessing their antioxidant behavior and understanding the mechanism of oxidative stress and its consequences. The choice of the method of synthesis should be based on the possibility to choose the conditions and parameters for obtaining CeO2 with controlled dimensions and a ratio of Се3+/Се4+ on their surface. In this study, CeO2 NPs are synthesized by precipitation in mixed water-alcohol solutions at constant pH = 9. The properties of obtained NPs are studied using various methods of physical-chemical characterization such as X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, and dynamic light scattering. The size of CeO2 NPs varied from 14 to 4.2 nm with increasing alcohol concentration, while the effect of constant pH during synthesis on the morphology of the particles was insignificant. The synthesized nanoparticles form highly stable aqueous suspensions since their zeta-potential is higher than + 40 mV. It is found that the ability of CeO2 NPs to self-stabilize is associated with the presence of hydrated Ce4+ ions on their surface. In vitro biological studies have shown that, regardless of particle size, CeO2 NPs have antioxidant potential, but smaller NPs with a higher percentage of Ce3+ on the surface had a more effective antioxidant effect. In addition, the size-depended activity of CeO2 NPs to inhibit the amyloid formation of insulin is demonstrated.

Structure and biological activity of particles produced from highly activated carbon adsorbent.

Over the recent years, carbon particles have gained relevance in the field of biomedical application to diminish the level of endo-/exogenous intoxication and oxidative stress products, which occur at different pathological states. However, it is very important that such carbon particles, specially developed for parenteral administration or per oral usage, possess a high adsorption potential and can remove hazard toxic substances of the hydrophilic, hydrophobic and amphiphilic nature usually accumulated in the blood due to the disease, and be absolutely safe for normal living cells and tissues of organism. In this work, the stable monodisperse suspension containing very small-sized (Dhydro = 1125.3 ± 243.8 nm) and highly pure carbon particles with an excellent accepting ability were obtained. UV-spectra, fluorescence quenching constant and binding association constant were provided by the information about conformational alterations in an albumin molecule in presence of carbon particles, about the dynamic type of quenching process and low binding affinity between carbon and protein. The later was confirmed by DSC method. In vitro cell culture experiments showed that carbon particles did not possess any cytotoxic effect towards all testing the normal cell lines of different histogenesis, did not show genotoxic effects and were absolutely safe for experimental animals during and after their parenteral administration. These observations may provide more information about how to develop a safe preparation of carbon particles for different biomedical applications, in particular, as a mean for intracorporeal therapy of various heavy diseases accompanied by the increased endogenous intoxication and the level of oxidative stress.

Biological activity of cerium dioxide nanoparticles.

Cerium dioxide nanoparticles (CeO2 NPs) with a high value of ζ-potential (≥30 mV) have been synthesized in reverse microemulsions and they are able to form the high-stable aqueous suspension without any additional stabilizers. It has been shown that the interaction of such CeO2 NPs with transport proteins, such as BSA, affects their molecular conformation and biochemical activity. The observed changes in the UV-absorbance spectrum and intrinsic fluorescence quenching of BSA molecule are indicative of the occurrence of structural changes caused by binding with the surface of CeO2 NPs. Low affinity between BSA and CeO2 NPs has been confirmed by differential scanning calorimetry (DSC). Moreover, CeO2 NPs can act as regenerative free-radical scavengers, and their antioxidant activity depends on the concentration. The positive charge of CeO2 NPs can be attributed to their low toxicity toward human malignant lymphocytes MT-4 and breast cancer cells MCF-7 however, the morphofunctional features of MCF-7 cells interacting with CeO2 NPs are indicative of the decrease in oncogenicity.

The effect of two formulations of carbon enterosorbents on oxidative stress indexes and molecular conformation of serum albumin in experimental animals exposed to CCl4.

The liver failure means inability to perform its normal synthetic, biotransformation and excretory functions. The disturbance of metabolic processes leads to the development of "metabolic endogenous intoxication" resulting in oxidative stress. Oxidative stress initiates the processes of oxidation of amino acid residues of blood plasma proteins causing the changes in their structure and functions. The effect of administration of highly activated porous carbonic enterosorbents on oxidative stress manifestations and molecular conformation of serum albumin in blood of experimental animals with acute liver failure induced by carbon tetrachloride (CCl4) needs to be investigated. Two forms of activated carbonic enterosorbents such as AC1 (primary beads with the range of diameters of 125-250 µm) and AC2 (secondary granules prepared from micronized AC1 having the mean particle size of ~1 µm) derived from phenol-formaldehyde resin were used in rat model with CCl4 intoxication. The total level of reactive oxygen species (ROS) in blood plasma, the activity of catalase (CAT) in blood hemolysates; the content of reduced glutathione (GSH) in liver homogenates, and the level of oxidative modification of proteins (OMP) such as aldehyde-dinitrophenylhydrazone (A-DNPH) and ketone-dinitrophenylhydrazone (K-DNPH) derivatives in blood plasma and liver homogenates were determined. In addition, the level of pro/antioxidant ratio in blood hemolysates and the content of lipid peroxidation product - malondialdehyde (MDA), in blood plasma and liver were determined. Melting thermograms of blood plasma proteins (BPP) and molecular conformation changes of serum albumin were analyzed by biophysical methods (differential scanning microcalorimetry and spectrofluorimetry). The extent of CCl4-induced oxidative damage in blood and liver of experimental animals was shown to be less expressed for AC1 in comparison with AC2 enterosorbent. However, AC2 used in the form of secondary granules positively influenced some biophysical properties of albumin molecule (temperature of melting, shape of melting endotherm and intrinsic fluorescence) after rats exposure to CCl4. In general, administration of both AC1 and AC2 led to the reduction of oxidative stress manifestations and partial restoration of native molecular conformation of serum albumin. These observations are promising in terms of achieving recovery of detoxification potential of organism after severe liver injury.

Effect of Primary and Secondary Beads of Carbon Enterosorbent on Haematological Parameters and Oxidative Stress Development Caused by Melphalan in Rats.

Background and Objectives: Side effects of anti-cancer drugs are usually accompanied by oxidative stress, including myelotoxicity. We evaluated the potential of oral highly activated micro-/macroporous carbon adsorbents (bulk density of 0.16 g/cm3, surface area calculation by Brunauer-Emmett-Teller model (SBET) > 2200 m2/g, derived from proprietary phenolic resin beads) to alleviate oxidative stress and myelotoxicity in rats. Materials and Methods: A single injection of cytostatic melphalan (L-PAM) at a dose of 4 mg/kg was used for modelling. Two forms of activated carbon were used: AC1-primary beads with the particle size range of 125-250 µm, and AC2-micronized AC1 with a mean particle size of ~1 µm. We measured haematological parameters white blood cells, red blood cells, platelet count, and haemoglobin level. Oxidative stress intensity was evaluated using the following markers: total levels of reactive oxygen species (ROS) in blood plasma; catalase activity (CAT) and pro-oxidant/antioxidant ratio in blood haemolysate samples; level of reduced glutathione (GSH) in liver tissues; oxidative modification of proteins, OPM (APHD, aldehyde-dinitrophenylhydrazone derivatives and KPHD, ketone dinitrophenylhydrazone derivatives) and malonic dialdehyde (MDA) in blood plasma and liver samples. Results: AC2 administration promoted significant myeloprotective effect: 1.5-fold increase in leukocytes, 2-fold in neutrophils, 1.5-fold in lymphocytes, and 1.23-fold in platelet count compared to the experimental Melphalan Group. At the same time, AC1 administration resulted in a slight increase in haematological parameters. Both ACs positively corrected important, but diverse, components of oxidative stress. They significantly reduced oxidative modification of blood and liver proteins (especially the AC1 form), normalized the level of reduced glutathione, pro-oxidant/antioxidant ratio and other markers. For some markers, such as ROS production in blood plasma, the use of enterosorbents resulted in non-significant a shift towards normal parameters. Conclusions: Oral activated carbon adsorbents reduce oxidative stress intensity and myelotoxicity; they can be promising means to combat the adverse effects of chemotherapy in clinical practice.

Heparin-cellulose-charcoal composites for drug detoxification prepared using room temperature ionic liquids.

We report novel heparin-cellulose-charcoal composites prepared using room temperature ionic liquids (RTILs) to enhance the biocompatibility and blood compatibility of activated charcoal beads while decreasing the size of their active pores.

Extraction of uraemic toxins with activated carbon restores the functional properties of albumin.

BACKGROUND: Previous work has demonstrated that a partial normalization of the conformation of albumin from uraemic plasma and a substantial restoration of its binding abilities can be achieved by extraction with activated charcoal. This is best achieved at pH 3, but exposure of whole plasma to this low pH leads to the loss of some essential components. METHODS: The melting curves and ligand-binding abilities of uraemic albumin have been investigated after extraction with a new generation of activated carbon at three pH values (7.2, 3.0 and 5.08). RESULTS: Albumin isolated from uraemic plasma had a characteristically increased melting temperature because of bound ligands. Extraction of uraemic plasma at pH 7.2, 5.08 and 3.0 induced low-temperature shifts of albumin thermo-adsorption maximum T1 of 1.4, 3.8, 2.4 degrees C and T2 of 0.8, 3.9 and 1.2 degrees C, respectively. Flow microcalorimetry data demonstrated a decrease in the ability of uraemic albumin to bind octanoate, phenol red, salicylic acid, warfarin and diazepam. Purification of uraemic plasma at pH 5.08 completely restored the binding affinity of albumin for all the marker ligands. CONCLUSIONS: Highly efficient activated carbons, with clinically feasible acidification of plasma, can remove strongly albumin-bound uraemic toxins. Investigation of the melting curve of the isolated albumin is a new biophysical way to monitor both its molecular condition and the extent of removal of protein-bound toxins by dialysis. The melting curve provides new qualitative and quantitative information about albumin in an analogous way to an electrocardiogram and the heart.

Albumin, bilirubin, and activated carbon: new edges of an old triangle.

The problem of interaction of human serum albumin (HSA), unconjugated bilirubin (UB) and high porosity activated HSGD carbons is investigated in this study. The decrease of UB to HSA molecular ratio by more than 300 times was demonstrated while the batch experiments in HSA-UB admixtures after contact with HSGD. HSGD carbons express extremely high activity for the removal of UB from HSA containing solutions (more than 100 mg of UB per 1 g of activated carbon). Ex-tempore albumin-coating of carbon surface decreases adsorbent capacity by bilirubin on 21%. At the same time ex-tempore albumin-coating of HSGD carbon surface as well as blood citratization prevent platelet and leukocytes loss and clotting inside of the column. Pharmacopoeia solution of HSA containing acetyl-tryptophan or octanoate used for albumin-coating of HSGD adsorbents, becomes ligand-free and rather more active in complexing with protein-bound substances. Combination of albumin-coated HSGD carbon as haemosorbent with HSA ligand-free solution as a transfusion media seems a new prospective modality of the extracorporeal removal of protein-bound toxins.

Effect of protein-bound uraemic toxins on the thermodynamic characteristics of human albumin.

The ability of albumin to bind drugs and other lipophilic organic acids is decreased in chronic renal failure by the accumulation of albumin-bound uraemic toxins such as hippuric acid, indoxyl sulphate and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF). This furan acid is the most highly bound and is not removed by haemodialysis. The inhibitory effects of these three uraemic toxins on the interaction of three marker ligands sodium octanoate (for medium chain fatty acids), salicylic acid and phenol red (bilirubin site/site I) with albumin have been investigated by differential scanning microcalorimetry and flow microcalorimetry. CMPF was the most potent inhibitor and its binding site coincided with that of bilirubin (site I). Indoxyl sulphate binds to the site for medium-chain fatty acids and tryptophan (site II) and hippuric acid, the weakest inhibitor, inhibited binding to the salicylic acid site.