Vol 16, No 1 (2021)

Obtaining of a primary cell culture of breast carcinoma is necessary both for the study of molecular and cellular mechanisms of tumor growth and for the selection of personalized therapy. However, when obtaining such culture, technical difficulties arise: poor adhesion to the substrate, increased growth of fibroblasts in culture, early aging, and others. The review describes the main options for culturing breast carcinoma cells - two-dimensional cultures, three-dimensional cultures, tissue sections, and also discusses methods for their preparation. The results of studies on changes in the receptor apparatus during cultivation and assessment of the effect of anticancer drugs on breast carcinoma cells in vitro are presented.

The use of modern antibacterial agents and antiseptics in modern orthopedics does not always prevent infectious complications. Currently, two-stage surgical treatment with the implantation of an antimicrobial spacer is common. This method increases the duration of treatment, causing additional surgical trauma. The use of bioresorbable material with additional antibiotic impregnation simultaneously with the rehabilitation of the infectious focus can be considered a promising direction for improving the effectiveness of treatment of chronic osteomyelitis and reducing the treatment time of this group of patients. The main group consisted of rabbits with an experimental model of osteomyelitis followed by rehabilitation and one-stage replacement of the bone defect with bioresorbable material impregnated with vancomycin (n=12), and in animals of the comparison group (n=12) - two-stage surgical intervention with a similar material. Morphological studies were performed on animals of both groups on the 45th and 90th days after operations with replacement of a bone defect with a bioresorbable material with vancomycin. On the 45th day after the operation, the intensity of the processes of formation of newly formed bone tissue and restructuring of the osteoarthritis replacement material was more pronounced in the main group (28,2 vs 23,5%). Two-stage treatment is characterized by a more pronounced formation of fibrous tissue, the area of which in dynamics increased by 1,4 times (from 27,6 to 39,3%), with a single-stage method of treatment, this indicator increased only by 2% (from 22,9 to 24,9%). The infectious process was stopped in all experimental animals. The effectiveness of one-stage surgical treatment seems to be determined by a faster onset of osteohistogenesis in the area of a local osteomyelic defect when biocomposite is administered immediately after the purulent focus is sanitized. In addition, the absence of repeated surgical trauma with the loss of additional bone volume when removing the cement spacer during two-stage treatment is likely to play a significant role.

To achieve greater clinical relevance of the newly discovered compounds, modern drug discovery requires disease-targeted assays based on human cells. The specific aim of this study was to design and develop a new cell-based assay for screening of compounds with IL-17A inhibitory activity. Human foreskin fibroblasts (HFF) were treated with IL-17A alone (experimental conditions I) or a mixture of IL-17A inhibitor netakimab and IL-17A (experimental conditions II). IL-17A - dependent production of inflammatory mediators IL-6, IL-8, MCP-1 was evaluated by ELISA (enzyme-linked immunosorbent assay). The study demonstrated the ability of HFF subcultured in vitro for a long time (>20 passages) to respond to IL-17A treatment by increased production of inflammatory cytokines IL-6, IL-8, MCP-1. Neutralization of IL-17A by netakimab (IL-17A inhibitor) resulted in a dose-dependent decrease of inflammatory cytokines production into cell growth medium. Thus, a new cell-based assay to evaluate the biological activity of Il-17A inhibitors has been developed and tested. The assay is based on the analysis of IL-17A-dependent production of inflammatory cytokines synthesized by human dermal fibroblasts. Netakimab has been shown to be a highly potent inhibitor of IL-17A.

Allergic inflammation is accompanied by stimulation of neutrophils with an increase in the formation of reactive oxygen species. The antioxidant effectiveness of some antihistamines is known, which reduces the risk of damage to surrounding tissues with the participation of these cells. Objective of the study: to determine the degree of various generations antihistamines influence on the death and enzymatic activity of neutrophils. The effect of the first antihistamines (diphenhydramine, clemastine) and second (lorata-dine, desloratadine) generations and the hormonal drug dexameth-asone on cell viability, the formation of active oxygen metabolites, enzyme activity, the amount of cationic proteins, and cytokine production by neutrophils was studied using the in vitro model. It was found that after exposure to loratadine at a dose of 2.5 |jg / ml, the number of viable cells was comparable (p = 0.001) with that in an intact culture. Found a stimulating effect of second generation antihistamines (loratadine, desloratadine) in low doses on the activity of NADPH-dependent oxide reductase. The form of neutrophil death depended on the type and dose of the drug; apoptosis was predominantly observed after cell contact with loratadine and desloratadine. Against the background of an increase in the activity of ATPase and myeloperoxidase after contact with diphenhydramine and clemastine (2.5 jg / ml), the largest number of neutrophils producing reactive oxygen species was revealed. Under the influence of desloratodine and clemastine, exocytosis of cationic proteins into the extracellular space and the lowest production of cytokines after contact with the latter were established. Thus, exposure to Hl-antihistamines, active both extra- and intracellular (diphenhydramine, loratadine), probably disrupted the metabolism of neutrophils, which led to an increase in their killer potential. Clemastine, acting mainly extracellularly, minimized the toxic effects of extracellular radicals, without affecting the production of intracellular oxidants involved in the regulation of neutrophil functions.

The problem of the restoration of the epithelial layer after various modifications of keratoplasty is of great fundamental interest. In this regard, new methods of induction of regeneration are being developed; one of the promising approaches in this area is the use of autologous blood derivatives with a high regenerative potential. Objective: to compare the effect of 3 blood derivatives serum, platelet-rich plasma and plasma rich growth factors on the culture of corneal epithelial cells. The study was carried out on cells of the epithelium of the human cornea of passage 3. To confirm corneal affiliation, cells were typed for characteristic cytokeratins. The dynamics of migration was assessed in the test for wound healing of the monolayer. Proliferation was assessed by the results of the formazan test. Plasma rich growth factors had the greatest stimulating effect on cell proliferation. There were no significant differences between groups in the rate of wound healing of the monolayer. It was found that, in comparison with the control, all stimulants shift the morphological phenotype of cells to a more mature side. As a result of the study, it was shown that all 3 types of tested blood derivatives are promoters of corneal re-epithelialization. The use of drugs obtained from blood can positively influence the processes of epithelialization in persistent epithelial corneal defects, which requires further study.

Schizophrenia is one of the most severe chronic relapsing mental diseases that significantly affect the level of social adaptation and quality of life of patients, often leads to their disability. Despite the success of modern psychopharmacology, achieving sustainable remission in schizophrenia remains a difficult task. The purpose of the study were the assessments of the safety and tolerability of intravenous administration of allogeneic AB0/ Rh-compatible mononuclear cord blood cells, as well as to study changes in cognitive performance in patients with schizophrenia in remission after treatment with umbilical cord blood cells. The study involved 30 patients with schizophrenia (men; average age 32,4 ± 9,7 years) in a state of hypochondria remission with a predominance of cognitive disorders against the background of prominent negative changes (F20.01-F20.04 according to ICD-1 0). Design is a prospective, placebo-controlled trial of efficiency and safety. The study consisted of 2 phases. In the pilot phase (3 months), the tolerability of a single cryopreserved concentrate of human cord blood injection containing mononuclear cells in a dose of 260± 20 million cells was estimated. The duration and severity of the effect was compared with placebo. In the clinical phase (48 months), patients received 4 injections of cord blood cell suspension in the same dose with intervals of 14 ± 3 days. The efficacy and safety of exposure were assessed using psychopathological, psychometric (scale of positive and negative symptoms of schizophrenia - PANSS) and psychological (The MATRIX Consensus Cognitive Battery) methods. The obtained results allow to conclude that the influence of human cord blood mononuclear cells on cognitive functions is realized due to the expressed metabolic (nootropic) and psychostimulating effects and restoration of normal neurotransmitters ratio. The effects are manifested in the form of activation of intellectual activity, acceleration of information processing, correction of memory functions, increase in the level of attention and vigilance, as well as a noticeable increase in "social intelligence” and, as a result, improvement in the quality of life. The effect of applying cord blood cells to enhance cognitive functions is characterized by resistance and duration of at least 4 years.