The morphogenetic properties of a collagen gel prepared by acetic acid extraction from the tendon sheaths (peritenons) of the paravertebral tendons of Wistar rats were studied. The gel was used as a substrate during in vitro cultivation together with mesenchymal stromal cells for 14 days in growth and osteogenic incubation media. It has been established that the collagen framework of the peritenon substrate is strengthened by increasing the connectivity of fibrillar nodes and is structured with the formation of lamellar and tangle formations. Sesamoid globules, penetrating into the substrate from the initial peritenon gel, remain inert during cultivation in the growth medium, but exhibit an increased ability to structure calcium phosphates in the osteogenic medium. The formation of cell-mediated structures occurs by directions of fibro, tendo-, ligament-, and osteogenic differentiation. The fibrogenic direction provides a structuring framework; the tenogenic direction provides the formation of embryonic tendons according to the mechanism of lateral assembly of collagen subfibrils on cell surfaces and their autonomization in the form of tendon filament primordia; the ligamentogenic direction provides structuring of collagen ribbons associated with tangles and elastic fibers; and the osteogenic direction provides the formation of lamellar, trabecular and nodular osteoid structures through intramembranous ossification, accompanied by activation of alkaline phosphatase and mineralization. The formation of enthesis predictors is the organization of commissures between mechanically different-phase components of osteoid structures and frame. A classification of taxonomic forms has been developed and a hypothesis has been proposed about the role of evolutionary tools in the structuring of the collagen framework in tissue cultures in vitro. A classification of taxonomic forms has been developed and a hypothesis has been proposed about the role of evolutionary tools in the structuring of the collagen framework in tissue cultures in vitro.

An important task of topical application of medicines in the treatment of eyes is to achieve a compromise between their effectiveness and safety. The development of new multifunctional local ophthalmic drug delivery systems and in vitro screening of potential medicinal eye products are key areas in solving this problem. In this study, primary in vitro screening of the effect of echinochrome (Ech), the carrageenan complex of echinochrome (CRG/Ech) and its liposomal form (CRG/Ech-Lip) was performed on cultured epithelial cells of the outer shell of the eyeball: conjunctival epithelial cells (Chang Conjunctiva, Clone 1-5c-4) and corneal epithelium human (HCE). The cell viability was assessed by their morphology and metabolic activity using light microscopy and MTT test methods. The direct dependence of the intensity of the cytotoxic effect of Ech on its concentration in the nutrient medium, the form of use, the cellular test system and the incubation time of cells was revealed. Ech in the form of an alcoholic solution in its final concentration of 0.1 mg/ml of the nutrient medium exhibits pronounced cytoxicity against both cellular test systems. The same final concentration of Ech in the nutrient medium, but already as part of the carrageenan complex of echinochrome (CRG/Ech), turned out to be critical only for the viability of corneal epithelial cells, the survival rate of conjunctival cells under these conditions was about 50 %. A high biocompatibility of the liposomal form of the carrageenan complex of echinochrome (CRG/Ech-Lip) with cells of both test systems and a stimulating cytoprotective effect against the cells of the conjunctiva epithelium was revealed.

— The effect of the Biosit-Sr Elkor glass-crystalline material on the early stages of morphogenesis of osteoid structures in cultures of mesenchymal stromal cells of bone marrow was studied. The maximum cultivation time was 28 days. Morphological investigations were performed with scanning electron microscopy. It was found that silicoaluminophosphate facilitated the generation of osteoid structures; enhanced the synthesis, compaction, and mineralization of fibrillar collagen; and activated matrix vesicles by hyperplasia, hypertrophy, and excretion. During cultivation, glass-crystalline material partially dissolved and disintegrated. Chemical substances with cross-linking, surface-active, and mineralizing properties came out into the nutrient medium. By means of surface-active agents, compacted collagen fibers were straightened and perforated, forming diversified lamellar structures, the precursors of bone plates. The mineralizing effect is mediated by increased homo- and heterogeneous nucleation of calcium phosphates and deposition of mineral concretions in differentiating cells and fibers of the collagen substrate, which accelerates the involution of cell culture. It was concluded that Biosit-Sr Elkor glass-crystalline material has a stimulating effect on the early stages of osteogenesis in vitro.

The secretome of human mesenchymal stem cells (MSCs) after osteogenic differentiation of FetMSC cells in vitro was obtained to develop tissue engineering technologies for bone tissue. For standard secretome samples a large number of FetMSC cells (7 × 108 cells) were cultured in serum-free conditioned medium (SFCM). All steps of cell processing were performed in the CompacT SelecT automatic system (Sartorius, United Kingdom). The SFCM was concentrated by ultrafiltration, then subjected to dialysis and dried in a vacuum rotary evaporator. The osteoinductive properties of SFCM concentrate (SFCMC) were tested on FetMSC cells by means of adding it to the growth medium at different concentrations. There were no changes in cell morphology during cultivation in the presence of SFCMC. Analysis of the transcription factors Runx2 and YAP1 (markers of osteogenic differentiation) expression using immunofluorescence staining and real-time polymerase chain reaction (RT-PCR) showed an increase in their level of expression in the presence of SFCMC. These findings demonstrate that it is possible to use SFCMC obtained from osteogenic MSCs to induce differentiation of other (undifferentiated) MSCs. The results are useful for the development of cell therapy products for bone restoration based on MSCs secretome.

The ability of the bone marrow multipotent mesenchymal stromal cells (MMSCs) of a patient with osteoarthritis to differentiate into the osteogenic and chondrogenic lineages under the influence of an electret-generated electric field created on the surface of anodic tantalum oxide was investigated. It has been shown that the synthesis of protein-differentiation markers in MMSCs is intensified in the presence of samples with anodic oxide in the electret state. The effect of electrets with uniform charge distribution over the surface was compared with linear charge distribution, MMSCs osteogenic differentiation was accompanied with enhanced expression of osteocalcin and type I collagen. These features were exhibited much stronger in cells exposed to electret specimens with a charge distribution on the surface close to linear.

The effect of the electric field of a medical electret on the basis of tantalum pentoxide (Ta2O5) on the viability of human fibroblasts, multipotent mesenchymal stromal bone marrow cells, osteocytes, and chondrocytes cultivated on the surface of tantalum orthopedic implants with an electret coating was studied in vitro. Using the methods of photocolorimetic analysis and scanning electron microscopy, difference in the metabolic activity and cell morphology, in the character of the cell attachment to the implants and distribution by their surface, associated with the value and character of the electric charge distribution on the surface of these implants and the type of cellular test systems, were detected. Unclear mechanisms of the effect of the electric field of medical electrets on the functional activity of different types of the organism cell and tissues dictate the need for further experimental developments and fundamental studies in this field.

Жизнеспособность клеток различных типов, культивируемых на поверхности медицинского электрета