The potential of a combination of decellularized tooth matrices and decellularized periodontal ligament in a collagen I hydrogel for periodontal regeneration was studied in a rat model of surgical periodontal defect. The results were evaluated using microcomputed tomography and histological and immunohistochemical methods on day 60. The bioengineered construct induced the formation of dental cement and new fibrous and bone tissues in the area of the periodontal ligament. This bioengineered construct can be considered as a possible bioscaffold for the restoration of the periodontal tissue.

The purpose of the research is to compare the accumulation of phenolic compounds of different species and varieties of mint, zoned in Central Russia against the background of plant infection by Meloidogyne incognita.Materials and methods. Plants were grown from cuttings in a growing experiment in open ground. Mentha × piperita L. (varieties: Tik-Tak, Orange, Minneola, Mojito, Mitchum, Chocolate), M. spicata L. (varieties Morocco, Crispa) and M. longifolia L. (Longifolia) were taken for the study. A month later, the rooted plants were infected at the rate of 1000 sp. infective larvae of M. incognita per plant. After 8 weeks leaves were fixed in ethanol. The total content of phenolic compounds (PC), phenylpropanoids, flavonoids and catechins was studied using a spectrophotometer. The determination of the total content of PС was carried out using the Folin-Cecolte reagent with measurement at 725 nm, phenylpropanoids – by direct measurement of optical density at 330 nm, flavonoids – by reaction with aluminum chloride at 415 nm, the total content of flavans (catechins - flavan-3-ols), their oligomeric forms – proanthocyanidins, as well as leukoanthocyanidins were assessed by reaction with vanillin at 500 nm.Results and discussion. It has been shown that the accumulation of phenols is related to the species of plants. The varieties Mentha × piperita L. in most cases contained more phenols than M. spicata L. and M. longifolia L. A significant number of PC was noted in the violet-colored varieties Mitchum, Chocolate and Orange. The total content of PC almost completely correlates with the content of their precursors – phenylpropanoids. In terms of the content of flavonoids, the Mitchum variety stands out noticeably, and in terms of the content of catechins, the Orange variety stands out. Nematode infection in most varieties causes a noticeable increase in the total accumulation of soluble PC, phenylpropanoids and flavans, but leads to a decrease in the content of flavonoids.

The purpose of the research is to study the effect of insect compost obtained as a result of the vital activity of insects of the Coleoptera order Ulomoides dermestoides on the quantitative and qualitative composition of soil nematodes of various ecological and trophic groups, as well as on the morphological and physiological state of plants and infection of tomato plants with root-knot nematodes.Materials and methods. Under laboratory conditions, soil containing a diverse fauna of nematodes was treated with 1% dry and 0.5; 0.75 and 1% aqueous solutions of biocompost. Insect compost was obtained by keeping the U. dermestoides on a dry nutrient mixture. Then a mixture of lawn grasses was sown in the ground. After 30 days, the composition of nematodes was analyzed. The ability of insect compost to suppress parasitic nematode species was studied using the tomato-knot nematode model system. Tomatoes were infected with Meloidogyne incognita at a rate of 500 larvae (J2) per plant and simultaneously treated with a 0.5% aqueous biocompost solution.Results and discussion. The insect compost U. dermestoides has an effect on quantitative and qualitative indicators in the community of soil nematodes, increasing the number of predatory and saprobiotic nematodes and displacing parasitic ones. And due to the content of various biologically active compounds, it affects the development of rootknot nematodes in tomato roots. When tomatoes are treated with an aqueous solution of insect compost, the infection score and the number of nematodes that penetrate the roots are reduced. The introduction of compost when growing a mixture of lawn grasses and tomatoes can improve the condition of the plants.

The efficacy of a commercial insecticidal preparation based on Paecilomyces fumosorosea, Pecilomycin PM116, against root-knot nematode (Meloidogyne arenaria) was evaluated on tomatoes. A comparison was made with biological (Nematophagin) and chemical (Vydate 5G) nematicides. Pecilomycin and Nematophagin did not suppress the development of plants, their weight and size were at the level of healthy controls. The analysis of photosynthetic pigments showed that the chlorophyll b level, the sum of chlorophylls and carotenoids when the root system was treated with Pecilomycin was comparable to healthy controls. Despite the fact that the plants treated with Pecilomycin were completely infected, their infection rate was 1.27 times lower than the controls, and single galls were only present on the roots. The treatment of the roots with Pecilomycin did not significantly affect the number and size of galls; the females in them were at the control level; however, the number of eggs in oothecae was 1.5 times less. Thus, Pecilomycin has an inhibitory effect on the root-knot nematode and a beneficial effect on the tomato plant. Further research and search for new strains of P. fumosorosea with high virulence against one or more pests opens up opportunities for the creation of new bionematicides.

The efficiency of bone tissue regeneration by decellularized tooth matrix, demineralized tooth matrix, and commercial xenograft Bio-Oss Spongiosa was compared on the model of a critical-size circular defect in the alveolar bone of the upper jaw of adult Wistar rats. The defect healing dynamics was assessed using histological, histomorphometrical, and immunohistochemical methods on days 30 and 60. In contrast to demineralized matrix and commercial xenograft, decellularized matrix induces the formation of the new bone tissue by day 60. Decellularized matrix can be considered as a biomaterial for cell-free tissue engineering for alveolar bone restoration in dentistry and maxillofacial surgery.

Extracellular vesicles (EVs) and, particularly, exosomes are becoming a promising material for “cell-free therapy” of many pathologic conditions. In the recent years therapeutic effects of mesenchymal stromal cells (MSCs) have been attributed to their secretory factors, including EVs. In this study we aim to investigate how EVs produced by MSCs of different tissue origin can influence myogenesis and fibrosis–the main processes that accompany skeletal muscle regeneration. Bone marrow, adipose tissue, intact muscle, and injured muscle-derived rat MSCs were obtained and cultured according to standard protocols. EVs were obtained by ultracentrifugation from the MSC-conditioned medium of cell passages 2 and 3. The effects of EVs were estimated on the in vitro models of myogenesis and fibrosis. Samples of isolated EVs contained nano-sized vesicles that carried some exosomal markers. Promyogenic microRNA were found in EVs from bone marrow and muscle MSCs. We found that MSC-derived EVs from all sources significantly increased the number of newly formed myotubes in myoblast culture in vitro and also reduced the number and size of fibrotic nodules in muscle fibroblast culture in vitro. Our results suggest that MSC-derived EVs indeed possess antifibrotic and promyogenic potentials. However, the role of microRNA in these processes is yet to be determined, and the effect of EVs on skeletal muscle regeneration is yet to be tested in vivo.

We studied the possibility of using decellularized tooth matrix as a scaffold to restore bone tissue. It was found that mesenchymal stem cells underwent spontaneous osteogenic differentiation on the decellularized tooth matrix, which makes it possible to use it as a natural allograft in the treatment of resorption of alveolar bone tissue.

The participation of mesenchymal stromal cells (MSCs) in the regeneration of muscle tissue was analyzed. It is shown that rat bone marrow MSCs do not show potency for myogenic differentiation in vitro under the influence of appropriate inducers and rarely fuse with myoblasts when co-cultured. However, the factors they release stimulate differentiation of myogenic cells. It is noted that in vivo, when combined with minced muscle tissue, MSCs enhance myogenesis and angiogenesis, and when injected into injured muscle, they improve the course of the recovery process, as well as the medium conditioned by them. The paracrine mechanism of the influence of MSCs on skeletal muscle regeneration has been established.

We analyzed the suitability of various collagen-based scaffolds for culturing and osteogenic differentiation of mesenchymal stromal cells (MSC). Decellularized and lyophilized swine intestinal submucosa (SIS) and porous collagen sponge made from reconstructed bovine derma (PCS) were the most effective in promoting MSC adhesion, survival, and growth. MSC from rat and mouse bone marrow and rat adipose tissue successfully adhered to the scaffold surface and penetrated into its deep layers. These scaffolds were also the most effective in inducing osteogenesis. These results indicate that microarchitectonics of PCS and SIS is optimal for support of MSC growth and osteogenic differentiation.

To clarify the organizing effect of Semax and HLDF-6 peptides on the kinetics of protein synthesis in hepatocytes, in addition to an in vitro study (Brodsky et al., 2019), the effects of these peptides in vivo have been studied. The circahoralian (ultradian) rhythm of protein synthesis, that is, a marker of the direct cell-to-cell communication, was investigated in rats of different ages. Peptides were injected intraperitoneally into young (2–3-month-old) or old (1.5–2-year-old) rats at a 50–100-µg/kg dose. Hepatocytes were isolated and sparse or dense cultures were established. In sparse cultures from young rats that received one or another peptide, the rhythm of protein synthesis was observed; in the cultures from rats of the same age that were injected with saline, no rhythm was found. In dense cultures of old rats after the action of the peptide, the amplitudes of the rhythm of protein synthesis did not differ from the rhythms observed in young rats; after saline injection, the rhythm amplitudes were twice as low. Injection of the peptide into adult rats that had previously received dopamine caused a protein synthesis rhythm characteristic for rats of this age; the administration of dopamine abolished the rhythm. The synchronizing effect of the peptides was maintained for at least 2 days after their single administration to the rat. The use of Semax or HLDF-6 is recommended to compensate for the disturbances of the kinetics of protein synthesis in humans in aging and pathology.

In the search for stable factors regulating direct cell–cell interactions and effects on the properties of cells in aging organisms, the regulatory peptides Semax and HLDF-6 were studied. The circahoralian rhythm of protein synthesis in cells in vitro served as a marker of cell–cell interactions. The peptides normalized the cell–cell interactions, which are greatly weakened during aging. It is shown that the peptides organize the protein-synthesis rhythm in primary rat hepatocyte cultures. The effect of the HLDF-6 peptide was realized via metabotropic glutamate receptors; the blockade of these receptors by the antagonist MCPG abolished the effect of the peptide. The protein kinase inhibitor H7 prevented the effect of the peptides on the protein-synthesis kinetics. Just as for other signaling factors, the activation of protein kinases in the case of the peptides regulates the key process of direct cell–cell interactions. The effect of a single signal of each of the peptides was retained for at least 1 day. Our data allow the peptides to be recommended for improving elderly people’s condition and block the factors that disorganize the protein-synthesis kinetics.

We studied the effect of bovine brain gangliosides, individual ganglioside GM1, and melatonin on the rate of wound closure under in vitro conditions and the effect of melatonin on the rate of wound healing under in vivo conditions. It was shown that bovine brain gangliosides and melatonin reliably increased cell migration in the experimental wound model. This effect was detected when the cell cultures were treated with the test preparations after wound infliction and when the cultures of human keratinocytes were pretreated before wounding. Analysis of the effect of melatonin on the rate of wound healing in vivo showed that melatonin accelerated this process, especially at the middle stages corresponding to the proliferation phase (days 3-6 after surgery). Histological analysis revealed intensification of epidermal cell proliferation at the edges of the wound starting from day 4 after surgery.

We studied the effect of mesenchymal stromal cells and conditioned media on healing of full-thickness skin wound in rats. Cell transplantation to the wound bed did not accelerate wound closing and had no effect on the severity of inflammation, but increased vasculariza - tion of the granulation tissue in 14 days after injury. After injection of conditioned medium to the wound, less pronounced inflammation or enhanced epithelialization was observed. The angiogenic effect was observed only after repeated administration of conditioned medium and was associated with slower regeneration, probably due to skin traumatization by repeated injections. At the same time, fetal skin fibroblasts stimulated angiogenesis only after transplantation in high doses and the medium conditioned by these fibroblasts had no effect on wound healing.

The kinetics of protein synthesis was investigated in primary cultures of hepatocytes from old rats in serum–free medium. The rats were fed mixed fodder supplemented with glutamic acid and then transferred to a regular mixed fodder. The amplitude of protein synthesis rhythm in hepatocytes isolated from these rats increased on average 2–fold in comparison with the rats not receiving glutamic acid supplement. Based on this indicator reflecting the degree of cell–cell interactions, the cells from old rats were not different from those of young rats. The effect was preserved for 3–4 days. These results are discussed in connection with our previous data on preservation of the effect of single administration of gangliosides, noradrenaline, serotonin, and other synchronizers on various cell populations. In contrast to the other investigated factors, glutamic acid is capable of penetrating the blood–brain barrier, which makes its effect possible not only in the case of hepatocytes and other non–brain cells, but also in neurons.

The aim of this study was to analyze the changes that occur in the population of bone marrow mesenchymal stromal cells (MSCs) during the individual development of an organism. For this purpose, the basic characteristics of MSCs (the content of clonogenic cells, immunophenotype, and potencies to differentiate in vitro and in vivo) in the prenatal, early postnatal, and late postnatal ontogeny of the rat were compared. It is shown that the cloning efficiency of bone marrow MSCs in 10-day-old and adult rats is comparable and hundreds of times smaller than that of bone cells of 20-day-old fetuses with a bone marrow rudiment. The activity of alkaline phosphatase, a marker of osteogenic cells, was found in the majority of colonies formed by MSCs of postnatal bone marrow but not by the fetal bone. By the CD90 expression and potencies for in vitro adipogenesis, the stromal cells from the fetal bone and bone marrow of 9- to 10-day-old rats were comparable with those of the mature bone marrow MSCs but differed from them by the small number of CD73-bearing cells and a weaker ability to osteogenesis in an inductive environment. The analysis of the fate of MSCs from the studied sources after their transplantation to adult rats showed that their ectopic transplantation as part of tissue fragments into the kidney results in the formation of bone tissues and hematopoietic stroma. In diffusion chambers with MSCs that were precultured in vitro, transplantation into the peritoneal cavity led to osteogenesis and chondrogenesis. However, no significant differences in the potencies of bone marrow MSCs for differentiation in vivo depending on the developmental stage have been found. Thus, during ontogeny, bone marrow MSCs enhance the expression of CD73 and the ability to osteogenesis in vitro, whereas the expression of CD90 and the potencies for adipogenesis in induction medium and differentiation in different directions in vivo do not change significantly.

Dense cultures of hepatocytes from old rats (~2 years old, body weight 530-610 g) are different from similar cultures of hepatocytes from young rats by the low amplitude of protein synthesis rhythm. Addition of glutamic acid (0.2, 0.4, or 0.6 mg/ml) into the culture medium with hepatocytes of old rats resulted in increase in the oscillation amplitudes of the protein synthesis rhythm to the level of young rats. A similar action of glutamic acid on the protein synthesis kinetics was observed in vivo after feeding old rats with glutamic acid. Inhibition of metabotropic receptors of glutamic acid with α-methyl-4-carboxyphenylglycine (0.01 mg/ml) abolished the effect of glutamic acid. The amplitude of oscillation of the protein synthesis rhythm in a cell population characterizes synchronization of individual oscillations caused by direct cell–cell communications. Hence, glutamic acid, acting as a receptor-dependent transmitter, enhanced direct cell–cell communications of hepatocytes that were decreased with aging. As differentiated from other known membrane signaling factors (gangliosides, norepinephrine, serotonin, dopamine), glutamic acid can penetrate into the brain and thus influence the communications and protein synthesis kinetics that are disturbed with aging not only in hepatocytes, but also in neurons.