Determination of the state of metabolism is an urgent problem in sports medicine. Biochemical and hematological blood parameters reflect the predominance of either anabolic or catabolic processes in metabolism, which depend on the intensity and duration of training. This study was conducted to determine the predominance of metabolic processes in 21 male rowing athletes (age, 22.55 ± 3.68 years; height, 189.62 ± 6.34 cm; and weight, 88.65 ± 8.55 kg). Monitoring of blood parameters and the level of intensity of training made it possible to establish a linear discriminant function, with which eight main blood parameters were determined for prediction of the metabolic state: creatinine (p < 0.001), uric acid (p < 0.001), urea (p < 0.001), testosterone (p < 0.001), alkaline phosphatase (p < 0.001), albumin (p < 0.05), total calcium (p < 0.05), and total protein (p < 0.05). The reliability of the developed prediction of metabolic activity of athletes using linear discriminant functions was confirmed by a strong positive correlation (r = 0.88; p < 0.001) with the results of assessing the activity of the neuroendocrine system. The accuracy of determining the metabolic state was 91.8

Purpose: To evaluate the frequency and spectrum of chromosome aberrations under X-Ray exposure at doses of 80, 250, and 1000 mGy in a human multipotent mesenchymal stromal cell (MMSC) cell line during long-term cultivation. Material and methods: MMSCs were isolated from human gingival mucosa by an enzymatic method and cultured in a serum-free medium. The presence of surface antigens was determined using the method of flow cytometry. The ability of the cell line to differentiate in the osteogenic, adipogenic, and chondrogenic directions was studied using induction media. Authentication was performed by genotyping of polymorphic STR loci, cytogenetic analysis was performed by multicolor fluorescent in situ hybridization (mFISH). Irradiation was carried out on an X-ray biological unit RUB RUST-M1 (Russia) at a dose rate of 40 mGy/min, a voltage of 100 kV, and a current of 0.8 mA. Results: At the first passage after irradiation, a statistically significant increase in the frequency of non-clonal CA compared with the control was recorded at a dose of 80, but not 250 and 1000 mGy. At the late stages of cultivation, the average frequency of breaks per chromosome in the group of non-irradiated cells did not differ from the values obtained after irradiation at doses of 80, 250, and 1000 mGy (p > 0.05). However, in MMSCs irradiated at a dose of 80 mGy, damage occurred more often in pairs of chromosomes 6 and 10, and at a dose of 1000 mGy, in a pair of chromosomes 9. A single irradiation of MMSCs in vitro did not affect the growth and progression of MMSCs characteristic of the studied primary cell line, of clonal cells with chromosome translocations and monosomy X, but led to an increase in the representation of a clone with tetrasomy 8. The total number of random clones with chromosome translocations that arose de novo increased after irradiation at a dose of 1000 mGy. Conclusion: Minor fluctuations in the proportion of cells with non-clonal CA, depending on the dose received in the early stages after irradiation (passage 1–4), disappeared at the later stages of cultivation (passage 8–14). There were no differences in mean frequencies between irradiated and non-irradiated MMSCs, but after irradiation, damage to some chromosomes could occur more frequently than others. A single X-ray irradiation of MMSCs can promote the growth and progression of primary pathological cytogenetic clones, regardless of the dose received, as well as an increase in the total number of de novo cell clones with chromosomal translocations that have arisen. A single X-ray irradiation of MMSCs can promote the growth and progression of primary pathological cytogenetic clones, regardless of the dose received, as well as an increase in the total number of de novo cell clones with chromosomal translocations that have arisen.

Purpose of the study. The aim of this work was to study the effect of conservation in various cryoprotectants on the biomechanical and morphological properties of the umbilical cord (UCA) arteries.Materials and methods. The material for this work was 12 umbilical arteries, which were divided into 4 study groups: (1) native UCA – control without freezing; (2) APC frozen in 5 % dextran with the addition of dimethyl sulfoxide (DMSO) to 10 % and 7.5 % human serum albumin (HSA); (3) APC frozen in glycerin and (4) UCA frozen in propanediol. For all groups, morphological and biomechanical studies were carried out. The statistical data were processed using the GraphPad Prism 9.2.0 program. Analysis of variance was used to find significant differences.Results. Freezing with 5 % dextran, 10 % DMSO and 7.5 % human albumin did not lead to significant changes in the morphology of vascular tissue compared to control. The conserved vascular tissue retains a moderate level of type I collagen, regardless of the method of conservation. The presence of statistically significant differences between the control and frozen UCAs was shown for Young's modulus (p < 0.05; 6.9 [6.8; 7.0] for the native artery; 2.3 [2.0; 2.6] for the artery preserved with the use of dextran, DMSO and human albumin; 2.3 [1.4; 3.2] for the artery preserved in glycerin and 0.8 [0.6; 1.0] – in propanediol, respectively).Conclusion. 1) Freezing and storage of UCA at 5 % dextran, 10 % DMSO and 7.5 % HSA does not lead to a significant change in the morphology of vascular membranes and cell nuclei; 2) The preservation of type I collagen according to the results of IHC is qualitatively reduced from a conditional indicator of +2 to +1 for frozen agroindustrial complexes compared with control ones; 3) Conservation leads to a significant decrease in the modulus of elasticity of the walls of the agroindustrial complex compared with control vessels (p < 0.05).

Purpose: Study of chromosomal aberration yields with a three-color FISH method in the peripheral blood lymphocyte cultures of patients with breast cancer before and after local irradiation. Material and methods: Venous blood was obtained from 24 patients aged from 33 to 79 years which suffering from cancer of the left or right breast after the implementation of a sectoral or radical resection of an affected organ with subsequent local adjuvant distant radiation therapy (DRT) to the scar area. Also it could be preceded by adjuvant polychimotherapy (PCT). A linear accelerator of the Trilogy system based on the Clinac iX platform was used for local irradiation. SOD was installed at 50 Gy per 25 fractions (daily 2 Gy). venous blood sampling produced to PCT and/or DRT and after 1 and/or 3 months after irradiation. A set of DNA probes to 1, 4 and 12 pairs of chromosomes was used for three-color FISH staining. Results: Levels of translocations, dicentrics and acentrics (per 100 cells) in peripheral blood lymphocyte cultures of patients up to DRT did not differ significantly with and without PCT and were amounted to average 0.63 ± 0.10; 0.10 ± 0.04 and 0.17 ± 0.06, respectively. The frequency of these chromosome rearrangements significantly increased after radiation therapy by an average of up to 4.62 ± 0.46; 1.48 ± 0.19 and 0.95 ± 0.14. The differences between 1 and 3 months of taking material after irradiation were absent. One of the patients, unlike the rest, was cytogenetically examined 11 years after a similar local irradiation, and the frequencies of translocations and dicentrics in her lymphocyte culture were 4.56 and 0.19 (per 100 cells), respectively, while they were varied from 1.63 to 10.00 for translocations and from 0.27 (0 in one patient) to 4.06 for dicentrics for other patients. Conclusion: When using a three-color FISH method there was a significant increase in the frequencies of translocations, dicentrics and acentrics in peripheral blood lymphocyte cultures of patients after DRT of breast cancer. At the same time there were more translocations than dicentrics and, apparently, they remain at the initial level for a long time. The frequency of translocations per 1 cell with translocations in the peripheral blood lymphocyte cultures in most cases exceeds 1 after local DRT of the breast cancer. The correlation between average physical doses to the whole body and cytogenetical estimates in individual patients was statistically insignificant.

Purpose: Evaluation of the effectiveness of the use of human decellularized amniotic membrane, both as an independent covering material and as a cell-free matrix for syngeneic regenerative cells of adipose tissue (stromal-vascular fraction – SVF, adipose tissue ‒ AT) in local radiation injuries (LRI) IIIb-IV severity in laboratory animals. Material and methods: 42 laboratory animals were studied. LRI modeling was carried out on an X-ray at a dose of 110 Gy. Animals were randomized randomly and divided into 6 groups depending on the type of treatment: 1st group (K) ‒ control group animals after irradiation did not receive specific treatment; the 2nd group (Gl) ‒ after irradiation, medical glue BF-6 was applied to the ulcer surface on the 21st day; 3rd group (Am) ‒ animals after irradiation on the ulcer surface was applied decellularized amniotic membrane, fixed with interrupted sutures on the 21st day; 4th group (Am-Gl) ‒ animals after irradiation on the ulcer surface was applied decellularized amniotic membrane, fixed with medical adhesive BF-6 for 21 days; 5th group (SVF-Gl) ‒ after irradiation, the SVF AT at a dose of 0.4×106 cells was applied to the ulcer surface after irradiation, followed by fixation with BF-6 medical glue for 21 days; the 6th group (Am-SVF) ‒ after irradiation, SVF AT was applied to the ulcer surface at a dose of 0.4×106 cells under the decellularized amniotic membrane, fixed with interrupted sutures on the 21st day. Results: On the 112th day, complete healing of the ulcer was observed in 100 % of animals in the Am-Gl group, in 70 % in the Am and Gl groups. There was no complete healing of the ulcer in the SVF-CGl and Am-SVF groups. The greatest reduction in the area of the total altered skin from 21 to 112 days of the experiment was noted in the groups Gl-SVF (34.7 %), K (31.6 %), Am-SVF (30.7 %). In the Am-Gl and Am groups, a reduction in the area of the total altered skin was recorded by 24.6 % and 14.7 %, respectively. In the GL group, the reduction in the area of the total altered skin was the smallest, by only 13.5 %. Conclusion: The use of a decellularized human amniotic membrane fixed with medical adhesive BF-6 can be considered as a promising method for the conservative treatment of LRI of the skin.

Objective. Isolation of stem cells (SCs) from different parts of the placenta and description of their biological characteristics. Materials and methods. The placenta was obtained from parturient women aged 22 to 39 years (mean age was 29.0 ± 3.7 years) with a normal singleton pregnancy resulting in normal delivery and with written informed consent obtained at 37–41 weeks of gestation after non-invasive (n = 32) or operative (n = 2) delivery. Wharton’s jelly from the umbilical cord and the fetal side of the placenta, the chorionic plate with chorion frondosum, were isolated and cultured to obtain mesenchymal stem cells (MSCs). Morphological evaluation of cell culture, differentiation, immunological characterization (CD34, CD45, CD90, CD105, CD73, HLA-DR (BD Biosciences and Becman Coulter, USA)), assessment of viability using 7-ADD dye on the FACSCanto II flow cytometry facility (Becton Dickinson CA, USA), cell growth, sterility control, analysis of STR (Short tandem repeat) loci polymorphism for identification, karyotyping, and cryopreservation of cell cultures were carried out. Results. Primary cell cultures were isolated from the chorionic plate with chorion frondosum and Wharton’s jelly of the umbilical cord in 100% of cases. MSCs had high proliferative activity and viability (98.0 ± 1.2% 7-ADD) throughout the whole period of cultivation. The growth rate of MSCs from the chorionic plate at P3, P4, and P5 was higher compared to that of MSCs from the Wharton’s jelly during cultivation, p ≤ 0.05. Immunophenotype corresponded to the requirements of International Society for Cellular Therapy: high expression of MSC markers (CD73, CD90, CD105) was detected; hematopoietic and lymphocytic markers were absent (CD34, CD45, HLA-DR). The potential of cells to differentiate in the mesenchymal direction (chondrocytes, osteocytes, adipocytes) was confirmed in all obtained MSCs. In 95% of cases, the cells were of fetal origin. The karyotype of the examined MSC cell lines was normal: 46, XX (female) or 46, XY (male). All cell cultures were tested, cryopreserved, and placed in a biobank. Conclusion. Isolation and evaluation of biological characteristics of human placental stem cells is of great interest and is a promising direction in regenerative medicine due to the simplicity of placenta sampling, the absence of ethical problems, the ability to quickly obtain and accumulate the necessary amount of cell material with a stable genotype and given biological characteristics. Key words: mesenchymal stem cells, placenta, immunophenotype, STR loci polymorphisms, karyotype

Introduction. Urethral stricture is a complex and urgent problem in operative urology. The main problem in the treatment of extended structures of the posterior urethra is the inability to form an adequate urethral site for augmentation urethroplasty with the common buccal graft, which has a priority in the treatment of penile strictures. The use of tissue acellular matrices may be promising in the development of reconstructive urology, which in the future will solve a number of problems associated with augmentation urethroplasty. The purpose of this article is to study the possibility of using a cell-free matrix of a donor artery as a free flat flap for stricture replacement urethroplasty on a model of laboratory animals (rabbits). Materials and methods. Donor blood vessels were Used, which were subjected to detergent-enzymatic perfusion decellularization. To assess the quality of the cell-free matrix, a histological study and an immunohistochemical study were performed. The cell-free flap of the donor artery was fixed to the protein envelope from the side of the simulated defect and posterior on-lay urethroplasty was performed.Results and discussion. The resulting matrix was characterized by the absence of detectable cell nuclei, preserved type I collagen, and a DNA content of no more than 50 ng / mg of tissue. In the postoperative period, normal motor activity of animals, normal urination, weight loss was not observed. The levels of C- reactive protein, creatinine, and urea in peripheral blood 5 months after surgery were within the normal range: 0.285±0.04839 mg / l, 93.5±8.057 mm / l, and 8.35±1.355 mm/l, respectively. If cystourethrography with the help of computer tomography data for stricture of the urethra is not revealed. During magnetic resonance imaging in the axial and sagittal projections, the patency of the urethra was indirectly confirmed. Conclusion. In a laboratory animal model, it was shown that the resulting cell-free flap has in vivo biocompatibility and can be used for replacement urethroplasty of posterior urethral strictures.

A cytogenetic study of cell lines of mesenchymal multipotent stromal cells (MMSCs) isolated from human bone marrow, adipose tissue, placenta, and gingival mucosa was carried out. Karyological analysis using two chromosome-staining methods (GTG and mFISH) showed a normal diploid karyotype of most MMSC lines up to the fifth passage of cultivation. Clonal chromosomal translocations were detected in 3 of the 22 investigated MMSCs, which is 14%. Analysis of 22 MMSC lines revealed a rather low frequency of cells with chromosomal aberrations (CAs) (within 5%). Among cells with chromosomal damage (3–5%) identified by mFISH analysis of 1440 metaphase plates, unstable chromosomal aberrations (fragments, dicentric chromosomes) account for about 65% of all damage, and stable (translocations) account for 35%. These results confirm the importance of cytogenetic studies of MMSC cell lines intended for medical purposes. Gathering data on chromosomal and karyotypic variability at different periods of cultivation will make it possible to determine the acceptable limits for the selection of genetically stable diploid MMSC cell lines in order to prevent undesirable consequences of their use for therapeutic purposes.

Purpose: to create a protocol for complex cryoconservation of blood vessels in polydimethylsiloxane to create tissue-specific matrices. Material and Methods. The material ofthe study was the iliac arteries from postmortem donors, seized in the multi-organ donation, rejected for unclaimed and disposed of later. Polydimethylsiloxane was used as a coolant (PDMS) with a viscosity of 5-25 centistokes (Spectroplast, Russia). Bench testing of cooling and heating ofthe blood vessel in PDMS, testing of heating and cooling of PDMS up to operating temperatures (-80°C) in different ways and variants of its thermal insulation, assessment of different cooling rates ofthe vessel during cryoconservation, modeling of 3 types of working chambers for cryoconservation, sterilization and defrosting of up to 10 vessels, modeling of cryoconservation and thawing of vessels in PDMS under different conditions, selection of substrates from silicone and plastic for uniform fixation of vessels which are of size or size in the working chambers or without them. Results. The developed protocol allows cryopreserved from 1 to 10 or more vessels of all basic sizes both immediately after removal and after transportation to the laboratory, to vary the cooling rate from 1.5 to 300°C/min and above, to be used with different levels of laboratory equipment, as with liquid nitrogen, dry ice, and only with a medical freezer. Conclusion. The developed protocol allows to apply it in cases of complex cryoconservation of blood vessels to create tissue-specific matrices and can be recommended for preclinical testing.

Aim: to investigate potential cryo- and radioprotective properties of a polydimethylsiloxane of low viscosity to provide safe and reliable cryopreservation and radiation sterilization of human cadaveric vascular allografts. Material and methods. A consecutive experimental set-up was carried out to establish polydimethylsiloxane (further — PDMS) protective endocellular properties. For this purpose a high-densitymesenchymal stem cells suspension was cryopreserved with 10% DMSO solution; PDMS exocellular protective properties were assessed by cyclic cryoconservation of ^femoral artery segments, while PDMS potential radioprotectiveproperties were assessed by gamma irradiation — induced sterilization of 3 cryopreserved femoral artery segments with gross length about 3cm, and for control grafts a slightly changed protocol with glycerin was used. In this study the dynamic viscosity of all cryopreserved vessels was analyzed with using specially designed experimental device, also electron scanning microscopy with lanthanide cells staining and routine hematoxylin — eosin cell staining were applied. Results. We have demonstrated a prominentexocellular protective effect of PDMS confirmed by a SEM and histological results, and also its mediated radioprotective effect, in particular because of its safer preliminary cryopreservation procedure. Conclusion. Use of PDMS for a cryopreservation and gamma sterilization showed its rationality, however requires further modification of protocols and additional researches.

Aim: to verify new techniques for human cadaveric vascular allografts cryopreservation, thawing and sterilization for the tissue engineering purposes. We use polydimethylsiloxane (PDMS) as a well-known, promising coolant. This allowed us to completely omit any cryoprotective or vitrifying solutions. Using of PDMS also makes possible an applying these allografts directly after freezing and decellularization and also it will also provide an opportunity to develop secure protocols of tissue— engineered vascular conduits cryopreservation. Matherial and methods. After mathematical modeling of cooling process and its validation the experiment for sealed (isolated) freezing at low temperature conditions of 30 femoral arterial segments has been conducted. The segments were at least 10 cm in length and taken from 15 cadaveric donors in the age of 65-85 years. The freezing process was carried out using the abovementioned coolant— PDMS, and then physico-mechanical properties of these allografts were evaluated with the special Instron machine. According to the results obtained, a modeling of their sterilization conditions was conducted (the grafts were freezed). Results. By physico-mechanical properties validation and restricted histological analysis it was shown that there was an accordance between freezed/thawed allografts properties and native vessels. Conclusion. The abovementioned approach for allografts cryopreservation and thawing was efficient enough for further work in this direction.

Data on application of decellularized scaffolds and tissue-engineered vascular conduits in the field of cardiovascular surgery have been analyzed, and also techniques for their procurement have been studied. For finding, selection and synthesis resources of research data from the systems Pubmed, ScienceDirect were used. The preference was given to the most informative, comprehensive and contemporary publications.