Laser Ablation-Generated Crystalline Selenium Nanoparticles Prevent Damage of DNA and Proteins Induced by Reactive Oxygen Species and Protect Mice against Injuries Caused by Radiation-Induced Oxidative Stress.

With the help of laser ablation, a technology for obtaining nanosized crystalline selenium particles (SeNPs) has been created. The SeNPs do not exhibit significant toxic properties, in contrast to molecular selenium compounds. The administration of SeNPs can significantly increase the viabilities of SH-SY5Y and PCMF cells after radiation exposure. The introduction of such nanoparticles into the animal body protects proteins and DNA from radiation-induced damage. The number of chromosomal breaks and oxidized proteins decreases in irradiated mice treated with SeNPs. Using hematological tests, it was found that a decrease in radiation-induced leukopenia and thrombocytopenia is observed when selenium nanoparticles are injected into mice before exposure to ionizing radiation. The administration of SeNPs to animals 5 h before radiation exposure in sublethal and lethal doses significantly increases their survival rate. The modification dose factor for animal survival was 1.2. It has been shown that the introduction of selenium nanoparticles significantly normalizes gene expression in the cells of the red bone marrow of mice after exposure to ionizing radiation. Thus, it has been demonstrated that SeNPs are a new gene-protective and radioprotective agent that can significantly reduce the harmful effects of ionizing radiation.

Fluoroplast Doped by Ag2O Nanoparticles as New Repairing Non-Cytotoxic Antibacterial Coating for Meat Industry.

Foodborne infections are an important global health problem due to their high prevalence and potential for severe complications. Bacterial contamination of meat during processing at the enterprise can be a source of foodborne infections. Polymeric coatings with antibacterial properties can be applied to prevent bacterial contamination. A composite coating based on fluoroplast and Ag2O NPs can serve as such a coating. In present study, we, for the first time, created a composite coating based on fluoroplast and Ag2O NPs. Using laser ablation in water, we obtained spherical Ag2O NPs with an average size of 45 nm and a ζ-potential of -32 mV. The resulting Ag2O NPs at concentrations of 0.001-0.1% were transferred into acetone and mixed with a fluoroplast-based varnish. The developed coating made it possible to completely eliminate damage to a Teflon cutting board. The fluoroplast/Ag2O NP coating was free of defects and inhomogeneities at the nano level. The fluoroplast/Ag2O NP composite increased the production of ROS (H2O2, OH radical), 8-oxogualnine in DNA in vitro, and long-lived active forms of proteins. The effect depended on the mass fraction of the added Ag2O NPs. The 0.01-0.1% fluoroplast/NP Ag2O coating exhibited excellent bacteriostatic and bactericidal properties against both Gram-positive and Gram-negative bacteria but did not affect the viability of eukaryotic cells. The developed PTFE/NP Ag2O 0.01-0.1% coating can be used to protect cutting boards from bacterial contamination in the meat processing industry.

Preparation and Investigation of Spherical Powder Made from Corrosion-Resistant 316L Steel with the Addition of 0.2% and 0.5% Ag.

The paper describes the production and study of spherical powder made from corrosion-resistant 316L steel with the addition of 0.2% and 0.5% Ag. The study of granulometric composition, morphology, fluidity and bulk density, phase composition, microhardness and impurity composition of the spherical powders was carried out. The study showed compliance of the spherical powders with the requirements for powders used for additive manufacturing. The fluidity of the powders was 17.9 s, and the bulk density was 3.76 g/cm3. The particles have a spherical shape with a minimum number of defects and an austenitic-ferritic structure. The study of the phase composition of ingots, wires and powders showed that the ingot structure of all samples consists of austenite. According to the results of studies of the phase composition of the wire, there is a decrease in γ-Fe and an increase in α-Fe and σ-NiCr in going from wire No. 1 to wire No. 3. According to the results of studies of the phase composition of the powder particles, there are three phases, γ-Fe, α-Fe, and Fe3O4. The study of microhardness showed a decrease in HV depending on the increase in silver. The hardness of the powder is lower than that of the ingot by 16-24% due to the presence of a ferritic phase in the powder. As a result of plasma spraying, an increase in residual oxygen is observed, which is associated with the oxidation of the melt during plasma dispersion. The amount of nitrogen and sulfur does not change, while the amount of carbon and hydrogen decreases, and the impurities content corresponds to the standards for corrosion-resistant steel. Qualitative and quantitative analysis of the silver content in the samples indicates that it was not affected by the stages involved in obtaining the spherical powder.

Investigation of Antibacterial Properties of Corrosion-Resistant 316L Steel Alloyed with 0.2 wt.% and 0.5 wt.% Ag.

The article is devoted to the study of melted ingots, plates rolled from them, and the resulting spherical powder made of corrosion-resistant 316L steel with the addition of 0.2 wt.% and 0.5 wt.% Ag. The study of antibacterial properties, microstructure, and distribution of silver concentrations, as well as qualitative analysis of silver content was carried out. The optimal mode of homogenization annealing of the ingot was 1050 °C for 9 h, which leads to the formation of an austenitic structure. It is shown that the addition of a small amount of silver does not affect the formation of the austenitic structure and silver is distributed evenly throughout the volume of the ingot. The austenitic structure also prevails in the plates after rolling. Silver is distributed evenly throughout the entire volume of the plate. It is noted that the addition of 0.2 wt.% Ag does not affect the strength, elongation, and microhardness of steel, and the addition of 0.5 wt.% Ag does not significantly reduce the strength of steel, however, all samples meet the mechanical characteristics according to the ASTM A240 standard. The qualitative chemical composition of samples made of corrosion-resistant steels was confirmed by X-ray fluorescence analysis methods. By the method of energy-dispersion analysis, the presence of a uniform distribution of silver over the entire volume of the powder particle was determined. The particles have a spherical shape with a minimum number of defects. The study of the antibacterial activity of plates and powder shows the presence of a clear antibacterial effect (bacteria of the genus Xanthomonas campestris, Erwinia carotovora, Pseudomonas marginalis, Clavibacter michiganensis) in samples No. 2 and No. 3 with the addition of 0.2 wt.% and 0.5 wt.% Ag.

Obtaining a Wire of Biocompatible Superelastic Alloy Ti-28Nb-5Zr.

Using the methods of electric arc melting, intermediate heat treatments, and consecutive intensive plastic deformation, a Ti-Nb-Zr alloy wire with a diameter of 1200 µm was obtained with a homogeneous chemical and phase (ß-Ti body-centered crystal lattice) composition corresponding to the presence of superelasticity and shape memory effect, corrosion resistance and biocompatibility. Perhaps the wire structure is represented by grains with a nanoscale diameter. For the wire obtained after stabilizing annealing, the proof strength Rp0.2 is 635 MPa, tensile strength is 840 MPa and Young's modulus is 22 GPa, relative elongation is 6.76%. No toxicity was detected. The resulting wire is considered to be promising for medical use.

Development of a Biocompatible PLGA Polymers Capable to Release Thrombolytic Enzyme Prourokinase.

The novelty of the work lies in the creation and study of the physical and biological properties of biodegradable polymer coatings for stents based on poly(lactic-co-glycolic acid) (PLGA). Polymer coatings are capable of prolonged and directed release of molecules with a high molecular weight, in particular, protein molecules of prourokinase (m.w. 54 kDa). A technology has been developed to create coatings having a relative elongation of 40% to 165% and a tensile strength of 25-65 MPa. Coatings are biodegradable; the rate of degradation of the polymer in an isotonic solution varies in the range of 0.05%-1.0% per day. The created coatings are capable of controlled release of the protein of prourokinase, while about 90% of the molecules of prourokinase retain their enzymatic activity. The rate of release of prourokinase can vary from 0.01 to 0.08 mg/day/cm2. Coatings do not have a short-term toxic effect on mammalian cells. The mitotic index of cells growing on coatings is approximately 1.5%. When implanting the developed polymers in animals in the postoperative period, there are no complications. Histological examination did not reveal pathological processes. When implanting individual polymers 60 days after surgery, only traces of PLGA are detected. Thus, a biodegradable composite mechanically resistant polymer capable of prolonged release of the high molecular weight prourokinase enzyme has been developed.

Biocompatibility of new materials based on nano-structured nitinol with titanium and tantalum composite surface layers: experimental analysis in vitro and in vivo.

A technology for obtaining materials from nanostructured nitinol with titanium- or tantalum-enriched surface layers was developed. Surface layers enriched with titanium or tantalum were shown to provide a decrease in the formation of reactive oxygen species and long-lived protein radicals in comparison to untreated nitinol. It was determined that human peripheral vessel myofibroblasts and human bone marrow mesenchymal stromal cells grown on nitinol bases coated with titanium or tantalum-enriched surface layers exhibit a nearly two times higher mitotic index. Response to implantation of pure nitinol, as well as nano-structure nitinol with titanium or tantalum-enriched surface layers, was expressed though formation of a mature uniform fibrous capsule peripherally to the fragment. The thickness of this capsule in the group of animals subjected to implantation of pure nitinol was 1.5 and 3.0-fold greater than that of the capsule in the groups implanted with nitinol fragments with titanium- or tantalum-enriched layers. No signs of calcinosis in the tissues surrounding implants with coatings were observed. The nature and structure of the formed capsules testify bioinertia of the implanted samples. It was shown that the morphology and composition of the surface of metal samples does not alter following biological tests. The obtained results indicate that nano-structure nitinol with titanium or tantalum enriched surface layers is a biocompatible material potentially suitable for medical applications.

Synthesis and Investigation of Photophysical and Biological Properties of Novel S-Containing Bacteriopurpurinimides.

Novel hybrid molecule containing 2-mercaptoethylamine was synthesized starting from O-propyloxime-N-propoxy bacteriopurpurinimide (dipropoxy-BPI), which was readily oxidized in oxygen atmosphere yielding the corresponding disulfide analogue (disulfide-BPI). Spectral, photophysical, photodynamic, and biological properties of compound were properly evaluated. Compounds bearing disulfide moiety can directly interact with glutathione (GSH), thereby reducing its intracellular concentration. Indeed, mice sarcoma S37 cell line was treated in vitro with disulfide-BPI, yielding a CC50 value of 0.05 ± 0.005 µM. A relatively high level of singlet oxygen was detected. It was demonstrated (by fluorescence) that the PS was rapidly accumulated in a cancer nest (S37) at a relatively high level after 2 h upon intravenous administration. After 24 h, no traces of the molecule were detected in the tumor mass. Moreover, high photodynamic efficiency was demonstrated at doses of 150-300 J/cm2 against two different in vivo tumor models, achieving 100% regression of cancer growth.

Photodynamic activity of the boronated chlorin e6 amide in artificial and cellular membranes.

Photodynamic tumor-destroying activity of the boronated chlorin e6 derivative BACE (chlorin e6 13(1)-N-{2-[N-(1-carba-closo-dodecaboran-1-yl)methyl]aminoethyl}amide-15(2), 17(3)-dimethyl ester), previously described in Moisenovich et al. (2010) PLoS ONE 5(9) e12717, was shown here to be enormously higher than that of unsubstituted chlorin e6, being supported by the data on much higher photocytotoxicity of BACE in M-1 sarcoma cell culture. To validate membrane damaging effect as the basis of the enhanced tumoricidal activity, BACE was compared with unsubstituted chlorin e6 in the potency to photosensitize dye leakage from liposomes, transbilayer lipid flip-flop, inactivation of gramicidin A ionic channels in planar lipid membranes and erythrocyte hemolysis. In all the models comprising artificial and cellular membranes, the photodynamic effect of BACE exceeded that of chlorin e6. BACE substantially differed from chlorin e6 in the affinity to liposomes and erythrocytes, as monitored by fluorescence spectroscopy, flow cytometry and centrifugation. The results support the key role of membrane binding in the photodynamic effect of the boronated chlorin e6 amide.

Novel photosensitizers trigger rapid death of malignant human cells and rodent tumor transplants via lipid photodamage and membrane permeabilization.

BACKGROUND: Apoptotic cascades may frequently be impaired in tumor cells; therefore, the approaches to circumvent these obstacles emerge as important therapeutic modalities. METHODOLOGY/PRINCIPAL FINDINGS: Our novel derivatives of chlorin e(6), that is, its amide (compound 2) and boronated amide (compound 5) evoked no dark toxicity and demonstrated a significantly higher photosensitizing efficacy than chlorin e(6) against transplanted aggressive tumors such as B16 melanoma and M-1 sarcoma. Compound 5 showed superior therapeutic potency. Illumination with red light of mammalian tumor cells loaded with 0.1 µM of 5 caused rapid (within the initial minutes) necrosis as determined by propidium iodide staining. The laser confocal microscopy-assisted analysis of cell death revealed the following order of events: prior to illumination, 5 accumulated in Golgi cysternae, endoplasmic reticulum and in some (but not all) lysosomes. In response to light, the reactive oxygen species burst was concomitant with the drop of mitochondrial transmembrane electric potential, the dramatic changes of mitochondrial shape and the loss of integrity of mitochondria and lysosomes. Within 3-4 min post illumination, the plasma membrane became permeable for propidium iodide. Compounds 2 and 5 were one order of magnitude more potent than chlorin e(6) in photodamage of artificial liposomes monitored in a dye release assay. The latter effect depended on the content of non-saturated lipids; in liposomes consisting of saturated lipids no photodamage was detectable. The increased therapeutic efficacy of 5 compared with 2 was attributed to a striking difference in the ability of these photosensitizers to permeate through hydrophobic membrane interior as evidenced by measurements of voltage jump-induced relaxation of transmembrane current on planar lipid bilayers. CONCLUSIONS/SIGNIFICANCE: The multimembrane photodestruction and cell necrosis induced by photoactivation of 2 and 5 are directly associated with membrane permeabilization caused by lipid photodamage.

Novel boronated chlorin e6-based photosensitizers: synthesis, binding to albumin and antitumour efficacy.

Chlorins, a class of plant porphyrins, are perspective as photosensitizing agents due to light absorption in the long wavelength spectral region and deeper photodamage of tissues. Aiming at optimization of antitumour properties of chlorins, we synthesized a series of boronated derivatives of chlorin e(6) and their complexes containing Zn(II), Pd(II) or Sn(IV). The compounds were synthesized by alkylation of amino or hydroxy derivatives of chlorin e(6) with 1-trifluoromethanesulfonylmethyl-o-carborane. Chlorin e(6) 13(1)-N-{2-[N-(o-carboran-1-yl)methyl]aminoethyl}amide-15(2), 17(3)-dimethyl ester (compound 5) formed complexes with serum albumin, a major porphyrin carrier. The binding constant of these complexes was approximately 4 times bigger than the respective value for the complexes of albumin with boron-free aminochlorin e(6). Compound 5 potently sensitized rat fibroblasts to illumination with monochromatic red light: >98% of cells were necrotic by 24h post-illumination with 1 microM of 5. This compound demonstrated high efficacy in photodynamic therapy of rat M-1 sarcoma. After PDT with 25mg/kg of 5 the residual tumours were significantly smaller than in animals subjected to PDT with equal concentration of boron-free aminochlorin e(6). No signs of general toxicity were detectable after PDT with 5. Thus, boronation can enhance the potency of chlorins in PDT, in particular, due to an increased binding to albumin. Our data expand the therapeutic applicability of boronated chlorins beyond boron neutron capture therapy; these agents emerge as dual efficacy photoradiosensitizers.

Boronated protohaemins: synthesis and in vivo antitumour efficacy.

The conjugates of porphyrin macrocycles with boron-containing polyhedra are under investigation as agents for binary treatment strategies of cancer. Aiming at the design of photoactive compounds with low-to-zero dark toxicity, we synthesized a series of carboranyl and monocarbon-carboranyl derivatives of protohaemin IX using the activation of porphyrin carboxylic groups with di-tert-butyl pyrocarbonate or pivaloyl chloride. The water-soluble 1,3,5,8-tetramethyl-2,4-divinyl-6(7)-[2'-(closo-monocarbon-carborane-1''-yl)methoxycarbonylethyl]-7(6)-(2'-carboxyethyl)porphyrin Fe(III) (compound 9) exerted no discernible cytotoxicity for cultured mammalian cells, nor did it cause general toxicity in rats. Importantly, 9 demonstrated dose-dependent activity as a phototoxin in photodynamic therapy of M-1 sarcoma-bearing rats. In animals injected with 20 mg kg(-1) of 9, the tumours shrank by day 3 after one single irradiation of the tumour with red laser light. Between 7 and 14 days post-irradiation, 88.9% of rats were tumour-free; no recurrence of the disease was detectable within at least 90 days. Protohaemin IX alone was without effect, indicating that boronation is important for the phototoxic activity of 9. This is the first study that presents the synthesis and preclinical in vivo efficacy of boronated derivatives of protohaemin as phototoxins. The applicability in photodynamic treatment broadens the therapeutic potential of boronated porphyrins beyond their conventional role as radiosensitizers in boron neutron capture therapy.