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Autoimmune diseases are characterized by dysregulation of immune responses and damage to healthy body tissues. Their complete cure remains elusive, and existing therapies are often accompanied by side effects. Recent studies have shown a signifi cant role of disturbances in the composition of the microbiome in the development of autoimmune reactions. Moreover, modulation of the microbiome through various therapeutic interventions represents a promising direction in the framework of complex therapy of the underlying disease. Extracellular vesicles, in particular exosomes, transport biologically active substances between cells, and a number of studies have shown their therapeutic effect in autoimmune diseases. However, the role of extracellular vesicles in modulating the microbiome remains poorly understood, and further research is needed to better understand their impact on the pathogenesis of autoimmune diseases and associated microbiome changes, as well as to develop new treatment strategies. The presented literature review, based on a study of English-language sources, examines the importance of the microbiota of different loci of the human body (intestines, skin, oral cavity) in the development of autoimmune diseases such as multiple sclerosis, psoriasis and Sjögren’s disease. The role of extracellular vesicles in modulating the microbiome during autoimmune diseases therapy is discussed.

Aim. To develop technology to create 3D-spheroid multipotent mesenchymal stem cells (MMSC) of limbal cadaveric human eyes, capable of safe and long-term secretion of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF).Materials and methods. MMSC were obtained by cultivation of limbal fragments, released from cadaveric donor human eye. Cultivation was carried out in DMEM/F12 medium, supplemented with L-glutamine, penicillin, streptomycin, amphotericin B, HEPES, insulin, dexamethasone and 10 vol.% FBS under standard conditions (5% СО2, 37 °C), medium change was performed every 3 days. To determine the phenotype of the received cell culture the method of immunophenotyping by marker proteins to MMSC (CD73, CD105, CD19, CD90, CD133) was used. Stimulation of neurotrophic factor secretion was performed via a twostep procedure. 3D-cell spheroids were created with the help of agarous plates for three groups of comparison, where group I was control group, spheroids of intact 2D-culture MMSK; group II – spheroids of previously induced 2D-culture MMSC; group III – spheroids of 2D-MMSC induced on the 1st day of cultivation. Cell cultures supernatants were selected in different periods for NGF and BDNF follow-up study by ELISA procedure.Results. Induction of 3D-spheroids of limbal MMSC promotes short-term increase of the level of BDNF and NGF, but further, the secretion of these factors significantly decreases. Induction leads to a change in the morphology of spheroids: loss of compactness and emergence of «fringed» (debris). Such changes indicate of frailty of received constructions. Spheroids from previously induced MMSC are capable of stable NTF secretion, but the level of secretion is much less as compared to the control group.Conclusion. 3D-cell culture of intact 2Dculture of limbal MMSC can be considered as cellular medication for a safe and long-term neuroprotection in optic neuropathy treatment.

Modern techniques of laser microsurgery of cell spheroids were used to develop a new simple reproducible model for studying repair and regeneration in vitro. Nanosecond laser pulses (wavelength 355 nm, frequency 100 Hz, pulse duration 2 ns) were applied to perform a microdissection of the outer and the inner zones of human bone marrow multipotent mesenchymal stromal cells (BM MMSC) spheroids. To achieve effective dissection and preservation of spheroid viability, the energy of laser pulses was optimized and adjusted in the range 7-9 μJ. After microdissection, the edges of the wound surface opened and the angular opening reached a value of more than 180°. The destruction of the initial spheroid structure was observed in the wound area, with surviving cells changing their shape into a round one. Partial restoration of a spheroid form took place in the first six hours. The complete structure restoration accompanying the reparative processes occurred gradually over seven days due to remodelling of surviving cells.

Background: The beginning of a new era of direct acting antivirals sets up its own rules, that is, to achieve the highest efficacy with the shortest duration of treatment. It is assumed that the use of the first generation of direct acting antivirals, similarly to interferon-free regimens, would allow for personalization of approaches to their prescriptions.Aim: To identify the most important parameters that can predict the greatest efficacy of triple antiviral therapy of 12 week duration in patients with chronic hepatitis C genotype 1.Materials and methods: The study included 204 patients with chronic hepatitis C virus (HCV) genotype 1 at an early stage of liver disease (METAVIR score F0-F2), who were either treatment-naive or had a history of relapse after standard of care antiviral therapy. In addition to routine work-up, all patients were screened for IL28B polymorphism; in the course of the treatment viral kinetics was assessed by an ultrasensitive polymerase chain reaction (PCR) (with lower limit of quantification of 12 IU/ml). Duration of the triple therapy (pegylated interferon-α2a, ribavirin and telaprevir) was reduced to 12 weeks if a rapid virological response was achieved; otherwise the patients continued their treatment in according with guidelines. Results: A complete rapid virological response was achieved in 174 patients (81.6%), in whom the duration of triple therapy was 12 weeks. According to the protocol, 25 patients with a partial rapid virological response continued their standard antiviral therapy for 12 weeks more. In those who achieved a rapid virological response, there was an association between IL28B-CC genotype at rs12979860 and maintenance of zero viremia at 12 weeks after termination of antiviral therapy (r = 0.38, p < 0.001). In all such patients there was a stable virological response at 12 weeks of the follow-up. Monitoring of viral load after 14 days of antiviral treatment was not predictive of its success. The preliminary results of a shortened (12 week) course of triple telaprevir-based viral therapy allowed to identify the most significant parameters of 100% efficacy, i.e., absence of the virus in blood at 12 weeks after termination of antiviral therapy. Conclusion: A 12 week course of triple telaprevir-based combination therapy is an optimal regimen for achievement of a stable virological response after 12 weeks of the follow-up in treatment-naïve patients with HCV genotype 1 or with a relapse after previous conventional antiviral treatment, who have IL28B – CC polymorphism, are at an early stage of liver disease and who achieve a rapid complete virological response confirmed by a highly sensitive PCR.

Purpose. To study prerequisites for a development of artificial cornea bioengineered design based on recombinant spidroin tissue matrix by behavior evaluation of 2D (planar) and 3D cell (threedimensional) cultures on its surface.Material and methods. We studied epithelioid and stromal primary cell cultures (MSC-L) received from the limbal zone of post-mortem donor eyes. Cells were seeded on Petri dishes and on cavities of cultural trays (Corning, USA). To get spheroid structures the cells after the second passage underwent the centrifuge and were seeded on agarous trays then were cultivated in thermostatic chamber (Cell-IQ, Chip Man Technologies, Finland) under standard conditions (37° C, 5% CO2). Control over cell growth and morphology in trays was conducted under inverted microscope CKX41 (Olympus, Japan). To count the cell quantity and their viability the automatic cell counter Countess (Invitrogen, USA) was used, to analyze the surface proteins expression the flow cytofluorimetry was applied. For matrices colonization we used the 3rd passage MSC-L and 7-day spheroids of MSC-L origin. To evaluate 2D and 3D cell cultures growth on the surface of membranous matrices of recombinant spidroin, to estimate its non-toxicity and adhesiveness the immunohistochemistry, light time-lapse microscopy (Cell-IQ, Chip Man Technologies, Finland), laser scanning confocal microscopy (FluoView FV10i, Olympus, Japan) and raster electronic microscopy (CamScan, Japan) were incorporated.Results. Few hours after cell seeding there was active cells’ attachment to the substrate. Attached cells were characterized by rounded, oval or polygonal ordonnance. 24 hours later bipolar elongated cells and islets of migrating epithelioid cells appearance were observed. In the incubation process under gravity force the spheroids were accumulated predominantly in the central zone of the matrix, 2 hours later an active migration of spheroids surface zone epithelioid cells was registered on the membrane. After 24 hours of incubation all seeded on the surface of membranous matrix cells possessed a mesenchyme-like phenotype. Spheroids had an ability to merge limitlessly, later we observed a new microtissue formation with epithelioid cells on the surface and mesenchyme-like cells in the central area. Both solitary spheroids and merger-derived microtissue contained epithelial and mesenchymal components as well as regularly organized fibrils of extracellular matrix.Conclusions. According to aforementioned data the development of artificial cornea bioengineered cell-tissue constructions based on the technology of 3D cell spheroids cultivation derived from multipotent stem cells of the limbus and spidroin matrix presents a promising prospect requiring a further profound investigation.