Publikationen

Biomechanical and biochemical changes in the muscle soleus of rats during imitation of hind limbs unuse were studied in the model of the Achilles tendon rupture (Achillotenotomy). Oral administration of water-soluble C60 fullerene at a dose of 1 mg/kg was used as a therapeutic agent throughout the experiment. Changes in the force of contraction and the integrated power of the muscle, the time to reach the maximum force response, the mechanics of fatigue processes development, in particular, the transition from dentate to smooth tetanus, as well as the levels of pro- and antioxidant balance in the blood of rats on days 15, 30 and 45 after injury were described. The obtained results indicate a promising prospect for C60 fullerene use as a powerful antioxidant for reducing and correcting pathological conditions of the muscular system arising from skeletal muscle atrophy.

The personal risks of infection, as well as the conditions for achieving herd immunity, are strongly dependent on an individual’s response to the infective agents on the one hand, and the individual’s reactions to vaccination on the other hand. The main goal of this work is to illustrate the importance of quantitative individual effects for disease risk in a simple way. The applied model was able to illustrate the quantitative effects, in the cases of different individual reactions, after exposition to viruses or bacteria and vaccines. The model was based on simple kinetic equations for stimulation of antibody production using different concentrations of the infective agent, vaccine and antibodies. It gave a qualitative explanation for the individual differences in breakthrough risks and different requirements concerning a second, third or further vaccinations, reconsidering different efficiencies of the stimulation of an immune reaction.

Noncovalent water-soluble nanocomplexes of C60 fullerene (C60) with chemotherapeutic drugs (Doxorubicin (Dox), Cisplatin (Cis), and herbal alkaloid Berberine (Ber)) were created. Their anticancer action toward various tumor cells was studied in vitro, addressing specifically their biological synergy, compared with the action of these drugs in the non-immobilized form. Different theoretical and experimental (SEM and AFM microscopy, UV-Vis, DLS, NMR and SANS spectroscopy, ITC calorimetry) methods were applied for getting insight into the nature of the nanocomplexes with drug molecules, as well as into the physical forces enabling stabilization of these complexes. Physicochemical mechanisms were proposed for drug interaction with C60. An enhancement of the toxic action of the created water-soluble C60-drug nanocomplexes toward cancer cells, compared to the action of free drug, was found. Specifically, the C60-Dox nanocomplexes demonstrated ˜3.5 higher cytotoxic potential in the leukemic cell lines (CCRF-CEM, Jurkat, THP1, and Molt-16) in comparison with free Dox in the nanometer range of concentrations. Besides, C60 doubled the intracellular level of the up-taken Dox, which also evidenced its function as a nanocarrier. The toxic effect of C60-Cis nanocomplex toward Lewis lung carcinoma (LLC) cells was shown to be higher with IC50 values 3.3 and 4.5 times at 48 h and 72 h, respectively, as compared to the IC50 of free drug. 12.5 [my] Cis had no effect on LLC cells' viability. The C60-Cis nanocomplex in Cis-equivalent concentration substantially decreased the viability of tumor cells, impaired their shape and adhesion, inhibited migration, and induced their accumulation in the pro-apoptotic sub-G1 phase of cell cycle. An induction of apoptosis by the C60-Cis nanocomplex was confirmed by the activation of caspase 3/7 and externalization of phosphatidylserine on the outer membrane of LLC cells after their double staining with the Annexin V-FITC/PI. The complexation with C60 promoted intracellular uptake of the Ber. An increase in C60 concentration in the C60-Ber nanocomplexes was accompanied by the elevation of their antiproliferative potential toward CCRF-CEM cells in the order: free Ber < 1:2 < 1:1 < 2:1.These findings suggest a universal potential of water-soluble pristine C60 as a unique nano-platform for the delivery of the chemotherapeutic drugs in cytotoxic effect of these drugs.

Introduction: 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).Methods: 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.Results: It was found that the maximum allowable cytotoxic concentration of C60 fullerene is 37.5 ± 3.0 [my]g/ml. The investigated C60FAS reduces the titer of coronavirus infectious activity by the value of 2.00 ± 0.08 TCID50/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.Conclusion: 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.

In this study the catalytic activity of different gold and bimetallic nanoparticle solutions towards the reduction of methylene blue by sodium borohydride as a model reaction is investigated. By utilizing differently shaped gold nanoparticles, i.e., spheres, cubes, prisms and rods as well as bimetallic gold–palladium and gold-platinum core-shell nanorods, we evaluate the effect of the catalyst surface area as available gold surface area, the shape of the nanoparticles and the impact of added secondary metals in case of bimetallic nanorods. We track the reaction by UV/Vis measurements in the range of 190-850 nm every 60 s. It is assumed that the gold nanoparticles do not only act as a unit transferring electrons from sodium borohydride towards methylene blue but can promote the electron transfer upon plasmonic excitation. By testing different particle shapes, we could indeed demonstrate an effect of the particle shape by excluding the impact of surface area and/or surface ligands. All nanoparticle solutions showed a higher methylene blue turnover than their reference, whereby gold nanoprisms exhibited 100% turnover as no further methylene blue absorption peak was detected. The reaction rate constant k was also determined and revealed overall quicker reactions when gold or bimetallic nanoparticles were added as a catalyst, and again these were highest for nanoprisms. Furthermore, when comparing gold and bimetallic nanorods, it could be shown that through the addition of the catalytically active second metal platinum or palladium, the dye turnover was accelerated and degradation rate constants were higher compared to those of pure gold nanorods. The results explore the catalytic activity of nanoparticles, and assist in exploring further catalytic applications.