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The aim of this study was to evaluate the antibacterial activity, quality and stability of creams (at 1 % concentration) prepared with synthesized silver(I) complexes: [Ag(Nam)2]NO3·H2O ( AgNam), [Ag2(HGly)2]n(NO3)2n (AgGly) (Nam – nicotin-amide, Gly – glycine) and silver(I) sulfadiazine (AgSD), which is commercially available. Antibacterial activity was evaluated by agar well diffusion method and in in vivo case. The pure silver(I) complexes as well as all three tested creams loaded with AgGly, AgSD and AgNam showed antibacterial potential. Moreover, the creams loaded with AgGly and AgNam showed higher antibacterial effects against S. aureus and B. subtilis than the cream loaded with AgSD. In terms of appearance, all cream samples were opaque and odourless, and no phase separation was observed. Creams were soluble in water (o/w emulsions) and they had a pseudoplastic behaviour. The pH of the creams was in the range of 4.87–5.75. No visible changes were observed in the case of commercially used AgSD cream during one month testing period at conditions –16 ± 1 °C; 6 ± 1 °C and 56 % relative humidity; 20 ± 1 °C and 58 % relative humidity and 40 ± 1 °C and 75 % relative humidity. However, creams containing AgGly and AgNam changed their colour depending on the tested conditions.

The effects of zinc sulphate on selected properties of L. plantarum CCM 7102 were tested in vitro. The resistance of lactobacilli to higher concentrations of ZnSO4 (up to 5000 mg Zn2+.l−1) in growth media was strain-dependent. Further studies were carried out on the most resistant strain of L. plantarum CCM 7102. While the addition of low concentrations of zinc sulphate into the growth media (< 100 mg Zn2+.l−1) did not influence the properties of L. plantarum CCM 7102, the concentrations of 100—500 mg Zn2+.l−1 stimulated: the growth rate, production of lactic acid, adhesion to porcine enterocytes and the inhibition of pathogens E. coli O8:K88+ent+, S. enterica and S. Typhimurium. Conversely, however, high concentrations > 500 mg Zn2+.l−1 inhibited these properties. The addition of zinc (250 mg Zn2+.l−1) did not affect the resistance to antimicrobials, low pH, and the resistance to bile salt was affected only weakly. Zinc-resistant probiotic Lactobacillus strains are suitable for use in feedstuffs with a higher content of zinc designed for the prevention of post weaning diarrhoea in pigs.

The aim of our study was to examine the effects of passive and active cell seeding techniques on in vitro chondrogenic differentiation of mesenchymal stem cells (MSC) isolated from rat bone marrow and seeded on porous biopolymer scaffolds based on polyhydroxybutyrate/chitosan (PCH) blends. This paper is focused on the distribution of the cells on and in the scaffolds, since it influences the uniformity of the created extracellular matrix (ECM), as well as the homogenity of the distribution of chondrogenic markers in vitro which ultimately affects the quality of the newly created tissue after in vivo implantation. The three types of cell-scaffold constructs were examined by: fluorescence microscopy, SEM, histology and quantitative analysis of the glycosaminoglycans after chondrogenic cultivation. The results demonstrated that the active cells seeded via the centrifugation of the cell suspension onto the scaffold guaranteed an even distribution of cells on the bulk of the scaffold and the uniform secretion of the ECM products by the differentiated cells.

This study investigated a possible relationship between exopolysaccharides (EPS) production and the resistance to bile salts and low pH in intestinal strains of Lactobacillus reuteri. The strains displayed a mucoid phenotype, when grown in the presence of 10 % sucrose. Scanning electron microscopy (SEM) revealed strands of exopolysaccharide linking neighbouring cells. The strains (except L. reuteri B1/1) produced EPS in the range from 15.80 to 650.70 mg.l−1. The strains were tested for tolerance to bile salts (0.15; 0.3 %) and low pH (1.5—2.0—2.5—3.0). The survival rate, after the treatment with artificial gastric and intestinal juices, was determined by flow cytometric analysis. The strains of L. reuteri that produced 121—650 mg.l−1 of EPS showed a significantly higher tolerance (P < 0.001) to the gastric juice at pH 3 and 2.5, throughout the entire exposure time, in comparison to the strains that produced less than 20 mg.l−1 of EPS. L. reuteri L26, with the highest production of EPS, exhibited the highest survival rate (60 %) at pH 2 after the 120 minutes of in-cubation and was able to tolerate pH 1.5 for 30 minutes. Higher production of EPS significantly (P < 0.001) increased the strains’ tolerance against the intestinal juice in the presence of 0.15 and 0.3 % bile salts and was time dependent. L. reuteri L26 showed the highest tolerance (P < 0.001) against 0.3 % bile salts. This investigation revealed a positive correlation between the EPS production and the resistance of intestinal L. reuteri to the stress conditions of the gastrointestinal tract (GIT).

Alginite is organic matter rich in humic substances and commonly found in nature, but despite that, the knowledge of its biological effects is limited. In our study we focused on monitoring the effects of alginite alone, as well as its effect as a carrier of probiotic lactobacilli on the cellular immune response in SPF mice after infection with Salmonella Typhimurium. Sixty six conventional SPF female mice of the Balb/c line were divided into 4 groups: 1. infection free negative control (NK) supplied neither alginite nor probiotic lactobacilli in the feed; 2. infection free alginite control (Alg) supplied feed with 10 % alginite; infected control supplied alginite in the feed but no lactobacilli; 3. infectious control (Alg + Sal) - animals infected with salmonella and supplied 10 % alginite in the feed but no lactobacilli;and 4. probiotic group (Lab + Alg + Sal) - animals infected with salmonella and administered 10 % alginite and Lactobacillus reuteri 2/6 in the feed. On day 21 of the experiments, the mice were bled and their mesenteric lymph nodes were taken after their death. The peripheral blood of the mice was analysed for the activity of phagocytes and the percentage of selected lymphocyte subpopulations was determined in the mesenteric lymph nodes and blood. The significantly highest phagocytic activity (FA) was noted in the infected group with alginite (Alg + Sal). The FA was significantly increased in groups Alg and Lab + Alg + Sal in comparison with the NK group. The highest engulfing ability of phagocytes (phagocytic index) was observed in the Lab + Alg + Sal group in comparison with other groups, but also in Alg group in comparison with NK. In the Lab + Alg + Sal group, we observed a significantly higher percentage of B-lymphocytes, CD4+CD8+ and natural killer T cells (NKT), but more significant impact on the numbers of subpopulations of lymphocytes was observed in the mesenteric lymph nodes, with the significantly highest proportions of CD4+CD8+lymphocytes and NK and NKT cells. Our results indicated immunomodulatory properties of alginite and L. reuteri 2/6 in salmonella infected mice, especially at the level of the innate immune system components. This activation of phagocytosis and NK cells can be used in the treatment of various infections.

The aim of this study was to identify beneficial bacteria with probiotic potential from kefir grains. The lactobacilli isolated from kefir grains were characterised as: Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus paracasei, and Lactobacillus kefiri. The strains Lb. plantarum 1Ž, Lb. paraplantarum S10, and Lb. paracasei 2Ž tolerated better the test gastric juice at pH 2 and 2.6 during 120 min of incubation in comparison with the strains Lb. kefiri. On the other hand, the strains Lb. kefiri were resistant to 0.3 % bile acid salts. The Lb. paracasei 2Ž showed the significantly highest survival (P < 0.001) at pH 2 in comparison with all other strains tested and was also able to tolerate 0.3 % concentration of the bile salts. All strains produced medium to strong biofilms on abiotic surfaces and inhibited the growth of selected potential pathogens with varying intensity. All kefir isolates were susceptible to the antibiotics tested and exhibited positive β-galactosidase activity with the exception of Lb. paracasei 2Ž which did not show any activity of undesirable enzymes, such as β-glucosidase and β-glucuronidase. Additional testing and validation of the biological properties and safety of the strain Lb. paracasei 2Ž under in vivo conditions are needed to confirm the prospective use of this strain in practice.

The flow cytometry method is used in many fields, not only scientific, but also clinical. In science, flow cytometry is used in immunology, molecular biology, microbiology or plant biology. In medicine, has its use, in the diagnosis of tumours, in reproductive and prenatal diagnosis, in transplants or in haematology. In our experimental work, we used this method to analyse colostrum and milk of sows. The aim of this study was to determine the number of somatic cells, the number of leukocytes and the number of T-lymphocytes. Colostrum samples were collected within 24 hours of delivery and then milk samples were collected at weekly intervals during 3 weeks. Statistical analysis revealed significant differences in the number of leukocytes (CD45+) in colostrum and milk. The most significant difference was noted between colostrum and milk in weeks 2 and 3 of the experiment (P < 0.0001), with the highest values found in colostrum. On the other hand, we found that the levels of helper T-cells (CD4+) and cytotoxic T-cells (CD8+) gradually increased over time (P > 0.05), with the highest values observed in the third week after farrowing. We can conclude that flow cytometry can be successfully used for the examination of somatic cells in sows’ milk.

American foulbrood is amongst the most dangerous diseases of the bee-family affecting many honeybee colonies worldwide. In countries of European Union, based on veterinary legislation, the bee colonies tested positive to American foulbrood are eradicated with high economic losses. It is therefore necessary to look for effective prevention, especially by the using of natural ingredients such as probiotics. In this study, we used lactobacilli isolated from digestive tracts of adult healthy honey bees and selected based on their good probiotic properties and ability to inhibit the growth of Paenibacillus larvae. These isolates were identified as Lactobacillus brevis and Lactobacillus plantarum. Night cultures of both strains were used for the preparation of probiotic suspensions and pollen was selected as an appropriate carrier for application of probiotic lacto-bacilli to the hives. Half a litre of pollen suspension was prepared for each hive. The suspension for experimental hives contained probiotic lactobacilli in concentration of 107–108 CFU.ml−1. Bees in control hives received pollen suspension without addition of probiotic lacto-bacilli. The suspensions were supplied to bees three times, once a week. Before and during administration of probiotics, samples of honey bees from each hive were taken every week and numbers of lactobacilli, P. larvae, enterobacteria and coliform bacteria were determined in their digestive tracts. Four weeks after the first administration of probiotic-pollen solution the numbers of lactobacilli were increased approximately by 0.5 log. Before starting this experiment, P. larvae (approx. 107 CFU.ml−1) were detected in digestive tracts of honey bees. In the experimental group after 3 weeks and in the control group after 4 weeks, no viable counts of P. larvae were found. The numbers of enterobacteria, coliform bacteria and Bacillus sp. decreased in both groups. During the experiment the health and condition of the hives were monitored. In addition, also monitored were: the development of the bee colony, the number of dead bees, the amount of hive debris, the aggressiveness of the bees, and the amount of honey spun; later during the autumn treatment also the fall of Varroa jacobsoni was noted. In the experimental and control groups, we recorded a decrease in the amount of hive debris. Only in the experimental group was a slightly above-average development of the bee colony, slightly above-average honey yields and a 70 % lower drop of Varroa mites. Based on these results, we assumed that the probiotic-pollen solution had a positive influence on the composition of microbiota in bee digestive tracts and it can increase resistance to P. larvae. It also had a positive effect on the health and condition of the bee colony. Probiotic-free pollen solution showed similar but weaker effects.

Alveolar echinococcosis (AE) caused by the larval stage of Echinococcus multilocularis is serious parasitic diseases associated with the host´s immunosuppression. The effects of human non-immune dialyzable leukocyte extract (DLE) on immune cells in blood and spleen and parasitic cysts weight in Balb/c mice after oral (PO), subcutaneous (SC) and intraperitoneal administration (IP) were compared. The reduction in cysts weight (p < 0.01) was recorded after PO route, whereas moderate reduction was found after SC and IP routes. The elevation of lymphoid populations in blood and spleen was found after PO administration (p < 0.01) in parallel with reduced myeloid population. Infection-elicited decline in B220+B cells was partially abolished by PO route, but DLE routes did not influence the CD3+ T cells. The proportions of CD3+CD4+Th lymphocytes were moderately upregulated, whereas CD3+CD8+Tc populations were reduced after all DLE routes (p < 0.01). PO administration increased CD11b+MHCIIhigh blood monocytes, CD11b-SigleF+ cell, but not CD11b+Si-glecF+ eosinophils in the blood, stimulated after SC and IP routes. DLE induced downregulation of NO production by LPS-stimulated adherent splenocytes ex vivo. Con A-triggered T lymphocyte proliferation was associated with the elevated IFN-γ production and transcription factor Tbet mRNA expression. The alleviation of Th2 (IL-4) and Treg (TGF-β) cytokine production by lymphocytes ex vivo paralleled with downregulation of gene transcription for cytokines, GATA and FoxP3. Reduction of myeloid cells with suppressive activity was found. The SC and IP routes affected partially the cysts weights, diminished significantly gene transcription, NO levels and Th2 and Treg cytokines production. Results showed that PO route of DLE administration was the most effective in ameliorating immunosuppression via stimulation of Th1 type, reducing Th2 and Treg type of immunity and CD3+CD8+Tc lymphocytes in the blood and spleens during E. multilocularis infection in mice.

The aim of this study was to determine the effects of sodium selenite on the level of oxidative stress and viability of spermatozoa in fresh bull ejaculate in in vitro conditions at different temperatures. Samples of the bull's ejaculates with a concentration of 7 × 105 spermatozoa per ml were placed into the commercial semen extender containing 0 (control), 1 (1Se), 3 (3Se) and 5 (5Se) µg.ml–1 of sodium selenite. The following analyses were performed by flow cytometry after 1, 3, 6, 8, 24, 48 and 72 hours of incubation at 4 °C and 37 °C. All analyses were carried out in triplicate. The level of oxidative stress at both temperatures were significantly lower in the experimental groups in comparison to the control group. However, a significant decline of live sperm concentration and a rise of damaged sperm concentration were recorded, especially in groups 1Se and 3Se in comparison to the control group. Only in group 5Se was there observed a positive effect on the damaged spermatozoa level in comparison with groups C, 1Se and 3Se at 4 °C. In conclusion, the applied concentrations of sodium selenite had a positive effect on the level of oxidative stress in all experimental groups, but mainly at concentration of 5 µg.ml–1 of sodium selenite, especially at 4 °C. However, the effect of selenium was not sufficient for improving the sperm viability.

Medicinal plants have been successfully used in the ethno medicine for a wide range of diseases since ancient times. The research on natural products has allowed the discovery of biologically relevant compounds inspired by plant secondary metabolites, what contributed to the development of many chemotherapeutic drugs. Flavonoids represent a group of therapeutically very effective plant secondary metabolites and selected molecules were shown to exert also antiparasitic activity. This work summarizes the recent knowledge generated within past three decades about potential parasitocidal activities of several flavonoids with different chemical structures, particularly on medically important flatworms such as Schistosoma spp., Fasciola spp., Echinococcus spp., Raillietina spp., and model cestode Mesocestoides vogae. Here we focus on curcumin, genistein, quercetin and silymarin complex of flavonolignans. All of them possess a whole spectrum of biological activities on eukaryotic cells which have multi-therapeutic effects in various diseases. In vitro they can induce profound alterations in the tegumental architecture and its functions as well as their activity can significantly modulate or damage worm´s metabolism directly by interaction with enzymes or signaling molecules in dose-dependent manner. Moreover, they seem to differentially regulate the RNA activity in numbers of worm´s genes. This review suggests that examined flavonoids and their derivates are promising molecules for antiparasitic drug research. Due to lack of toxicity, isoflavons could be used directly for therapy, or as adjuvant therapy for diseases caused by medically important cestodes and trematodes.

The metacestode stage of the tapeworm Mesocestoides vogae (M. vogae) has the ability of asexual growth in the peritoneal cavity of rodents and other intermediate hosts without restriction. Early immunological events have decisive role in the establishment of infection. In the present study we investigated the kinetic of myeloid and lymphoid cell populations and the proportions of cells undergoing apoptosis in peritoneal cavities of mice within the first month after oral infection with M. vogae larvae. Proportions of cell phenotypes and apoptotic cells were examined by flow cytometry and by microscopical analysis of cells following May/Grünwald staining and fluorescent stain Hoechst 33234, respectively. Total numbers of peritoneal cells increased and their distribution changed towards accumulation of myelo-monocytic cell lineage in the account of reduced proportions of lymphoid cells. CD4+ T cell subpopulations were more abundant than CD8+ and their proportions elevated within two weeks post infection (p.i.) which was followed by a significant decline. Expression level of CD11c marker on myelo-monocytic cells revealed phenotype heterogeneity and proportions of cells with low and medium expression elevated from day 14 p.i. along with concurrent very low presence of CD11chigh phenotype. Lymphoid cell population was highly resistant to apoptosis but elevated proportions of myeloid cells were in early/late stage of apoptosis. Apoptosis was detected in a higher number of adherent cells from day 14 p.i. onwards as evidenced by nuclear fluorescent staining. By contrast, cells adherent to larvae, mostly macrophages and eosinophils, did not have fragmented nuclei. Our data demonstrated that apoptosis did not account for diminished population of peritoneal lymphoid cells and substantial proportions of myeloid cells seem to be more susceptible to apoptotic turnover in peritoneal cavity of mice with ongoing M. vogae infection, suggesting their important role in the host-parasite interactions.

This study aimed to evaluate the immunomodulatory effect of the probiotic Limosilactobacillus reuteri L26 Biocenol TM (L26) and its purified exopolysaccharide (EPS) with respect to antiviral innate immune response. In our experiment, we used porcine epithelial IPEC-J2 cells as a model of the intestinal barrier in a homologous infection by porcine Rotavirus   A  strain OSU6 (RVA). The production of selected molecules of non-specific humoral immunity was evaluated at the mRNA level. The EPS alone significantly increased the level of interferon λ3 (IFN-λ3) mRNA in the non-infected IPEC-J2 cells ( P  < 0.001). We also tested whether the treatment of IPEC-J2 cells by L26 or EPS influences the replication of RVA by virus titration and real-time PCR. We found that a pre-treatment in combination with subsequent continuous stimulation has no influence on the RVA replication. However, both treatments significantly decreased the RVA-induced production of IFN-λ3 ( P  < 0.05) and the "SOS" cytokine interleukin 6 (IL-6; P  < 0.01), already at the transcription level. In addition, the EPS treatment resulted in significantly increased IL-10 mRNA in the RVA-infected cells. In summary, we assume an immunoregulatory potential of  L. reuteri L26 Biocenol TM and its EPS in the local intestinal antiviral immune response., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2024 Schusterova et al.)

Transplantation of faecal microbiota (FMT) is generally considered a safe therapeutic procedure with few adverse effects. The main factors that limit the spread of the use of FMT therapy for idiopathic inflammatory bowel disease (IBD) are the necessity of minimising the risk of infection and transfer of another disease. Obtaining the animal model of UC (ulcerative colitis) by exposure to DSS (dextran sodium sulphate) depends on many factors that significantly affect the result. Per os intake of DSS with water is individual for each animal and results in the development of a range of various forms of induced UC. For this reason, the aim of our study was to evaluate the modulation and regenerative effects of FMT on the clinical and histopathological responses and the changes in the bowel microenvironment in pseudo germ-free (PGF) mice of the BALB/c line subjected to chemical induction of mild, moderate and serious forms of UC. The goal was to obtain new data related to the safety and effectiveness of FMT that can contribute to its improved and optimised use. The animals with mild and moderate forms of UC subjected to FMT treatment exhibited lower severity of the disease and markedly lower damage to the colon, including reduced clinical and histological disease index and decreased inflammatory response of colon mucosa. However, FMT treatment failed to achieve the expected therapeutic effect in animals with the serious form of UC activity. The results of our study indicated a potential safety risk involving development of bacteraemia and also translocation of non-pathogenic representatives of bowel microbiota associated with FMT treatment of animals with a diagnosed serious form of UC.

Introduction: With growing significance in nervous system repair, mesenchymal stem cell-derived conditioned media (MSCCM) have been used in cell-free therapies in regenerative medicine. However, the immunomodulatory and neuroregenerative effects of MSCCM and the influence of priming on these effects are still poorly understood., Methods: In this study, by various methods focused on cell viability, proliferation, neuron-like differentiation, neurite outgrowth, cell migration and regrowth, we demonstrated that MSCCM derived from adipose tissue (AT-MSCCM) and amniotic membrane (AM-MSCCM) had different effects on SH-SY5Y cells., Results and Discussion: AT-MSCCM was found to have a higher proliferative capacity and the ability to impact neurite outgrowth during differentiation, while AM-MSCCM showed more pronounced immunomodulatory activity, migration, and re-growth of SH-SY5Y cells in the scratch model. Furthermore, priming of MSC with pro-inflammatory cytokine (IFN-γ) resulted in different proteomic profiles of conditioned media from both sources, which had the highest effect on SH-SY5Y proliferation and neurite outgrowth in terms of the length of neurites (pAT-MSCCM) compared to the control group (DMEM). Altogether, our results highlight the potential of primed and non-primed MSCCM as a therapeutic tool for neurodegenerative diseases, although some differences must be considered., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hudakova, Mudronova, Marcincakova, Slovinska, Majerova, Maloveska, Petrouskova, Humenik and Cizkova.)

In this study, we provide comprehensive analyses of mesenchymal stem cells (MSCs) isolated from three types of canine tissues: bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and amniotic tissue (AM-MSCs). We compare their morphology, phenotype, multilineage potential and proliferation activity. The BM-MSCs and AM-MSCs showed fibroblast-like shapes against the spindle shape of the AT-MSCs. All populations showed strong osteogenic and chondrogenic potential. However, we observed phenotypic differences. The BM-MSCs and AT-MSCs revealed high expression of CD29, CD44, CD90 and CD105 positivity compared to the AM-MSCs, which showed reduced expression of all the analysed CD markers. Similarly, the isolation yield and proliferation varied depending on the source. The highest isolation yield and proliferation were detected in the population of AT-MSCs, while the AM-MSCs showed a high yield of cells, but the lowest proliferation activity, in contrast to the BM-MSCs which had the lowest isolation yield. Thus, the present data provide assumptions for obtaining a homogeneous MSC derived from all three canine tissues for possible applications in veterinary regenerative medicine, while the origin of isolated MSCs must always be taken into account.

Due to the physiological complexity of the tumour, a single drug therapeutic strategy may not be sufficient for effective treatment. Emerging evidence suggests that combination strategies may be important to achieve more efficient tumour responses. Different immunomodulators are frequently tested to reverse the situation for the purpose of improving immune response and minimizing chemotherapy side effects. Immodin (IM) represents an attractive alternative to complement chemotherapy, which can be used to enhance the immune system after disturbances resulting from the side effects of chemotherapy. In the presented study, a model of CT26 tumor-bearing mice was used to investigate the effect of single IM or its combination with 5-fluorouracil (5-FU) on colon cancer cells. Our results highlight that the beneficial role of IM claimed in previous studies cannot be generalised to all chemotherapeutic drugs, as 5-FU toxicity was not increased. On the contrary, the chemotherapeutic anti-cancer efficacy of 5-FU was greatly compromised when combined with IM. Indeed, the combined treatment was significantly less effective regarding the tumour growth and animal survival, most probably due to the increased number of tumour-associated macrophages, and increased 5-FU cytotoxic effect related to kidneys and the liver.

Neurodegenerative diseases are common problem for companion animals. Due to the limited ability of injured axons to regenerate, innovative therapies combined with rehabilitation have been applied and evaluated. Among them, stem cells and their conditioned media implantation, which can ameliorate damaged tissue has been suggested as a promising treatment strategy. The main goal of our study was to characterize mesenchymal stem cells (MSC) derived from canine adipose tissue (AT-MSC) and umbilical cord (UC-MSC) and analyse effect of their conditioned media (CM) on neurite outgrowth of neural progenitor cells isolated from the brain cortex of neonatal rats. MSC from both sources showed high osteogenic and chondrogenic potential and expression of CD90 and CD29. Furthermore, both UC-MSCCM and AT-MSCCM stimulated neurite growth. Interestingly, this effect was more pronounced with UC-MSCCM when compared to AT-MSCCM in vitro, which may be related to the different content of neurotrophic factors included in the CM.

The aim of this study was to investigate the use of a standardized animal model subjected to antibiotic treatment, and the effects of this treatment on the course of dextran sodium sulphate (DSS)-induced colitis in mice. By decontamination with selective antibiotics and observation of pathogenesis of ulcerative colitis (UC) induced chemically by exposure of mice to various concentrations of DSS, we obtained an optimum animal PGF model of acute UC manifested by mucin depletion, epithelial degeneration and necrosis, leading to the disappearance of epithelial cells, infiltration of lamina propria and submucosa with neutrophils, cryptitis, and accompanied by decreased viability of intestinal microbiota, loss of body weight, dehydration, moderate rectal bleeding, and a decrease in the selected markers of cellular proliferation and apoptosis. The obtained PGF model did not exhibit changes that could contribute to inflammation by means of alteration of the metabolic status and the induced dysbiosis did not serve as a bearer of pathogenic microorganisms participating in development of ulcerative colitis. The inflammatory process was induced particularly by exposure to DSS and its toxic action on compactness and integrity of mucosal barrier in the large intestine. This offers new possibilities of the use of this animal model in studies with or without participation of pathogenic microbiota in IBD pathogenesis.

Spinal cord injury (SCI) involves nerve damage and often leads to motor, sensory and autonomic dysfunctions. In the present study, we have designed a clinical protocol to assess the feasibility of systemic delivery of allogenic canine bone marrow tissue-derived mesenchymal stem cell conditioned medium (BMMSC CM) to dogs with SCI. Four client-owned dogs with chronic SCI lasting more than six months underwent neurological and clinical evaluation, MRI imaging and blood tests before being enrolled in this study. All dogs received four intravenous infusions with canine allogenic BMMSC CM within one month. Between the infusions the dogs received comprehensive physiotherapy, which continued for three additional months. No adverse effects or complications were observed during the one, three and six months follow-up periods. Neither blood chemistry panel nor hematology profile showed any significant changes. All dogs were clinically improved as assessed using Olby locomotor scales after one, three and six months of BMMSC CM treatment. Furthermore, goniometric measurements revealed partial improvement in the range of joint motion. Bladder function improved in two disabled dogs. We conclude that multiple delivery of allogenic cell-derived conditioned medium to dogs with chronic SCI is feasible, and it might be clinically beneficial in combination with physiotherapy.

The articular cartilage is an avascular and aneural tissue and its injuries result mostly in osteoarthritic changes and formation of fibrous tissue. Efforts of scientists worldwide are focused on restoration of cartilage with increase in life quality of patients. Novel polymeric polyhydroxybutyrate/chitosan (PCH) porous 3D scaffolds were developed and characterized. The rat mesenchymal stem cells (MSCs) were seeded in vitro on PCH scaffolds by a simple filtration of MSCs suspension over scaffolds using syringe. The chondrogenesis of cell-scaffold constructs was carried out in supplemented chondrogenic cultivation medium. After 2 and 4 weeks of in vitro culturing cell-scaffold constructs in chondrogenic differentiation medium, the cartilage extracellular matrix components like glycosaminoglycans and collagens were identified in scaffolds by biochemical assays and histological and immunohistochemical staining. Preliminary in vivo experiments with acellular scaffolds, which filled the artificially created cartilage defect in sheep knee were done and evaluated. Cells released from the bone marrow cavity have penetrated into acellular PCH scaffold in cartilage defect and induced tissue formation similar to hyaline cartilage. The results demonstrated that PCH scaffolds supported chondrogenic differentiation of MSCs in vitro. Acellular PCH scaffolds were successfully utilized in vivo for reparation of artificially created knee cartilage defects in sheep and supported wound healing and formation of hyaline cartilage-like tissue.

Adult stem cells have become prominent candidates for treating various diseases in veterinary practice. The main goal of our study was therefore to provide a comprehensive study of canine bone marrow-derived mesenchymal stem cells (BMMSC) and conditioned media, isolated from healthy adult dogs of different breeds. Under well-defined standardized isolation protocols, the multipotent differentiation and specific surface markers of BMMSC were supplemented with their gene expression, proteomic profile, and their biological function. The presented data confirm that canine BMMSC express important genes for differentiation toward osteo-, chondro-, and tendo-genic directions, but also genes associated with angiogenic, neurotrophic, and immunomodulatory properties. Furthermore, using proteome profiling, we identify for the first time the dynamic release of various bioactive molecules, such as transcription and translation factors and osteogenic, growth, angiogenic, and neurotrophic factors from canine BMMSC conditioned medium. Importantly, the relevant genes were linked to their proteins as detected in the conditioned medium and further associated with angiogenic activity in chorioallantoic membrane (CAM) assay. In this way, we show that the canine BMMSC release a variety of bioactive molecules, revealing a strong paracrine component that may possess therapeutic potential in various pathologies. However, extensive experimental or preclinical trials testing canine sources need to be performed in order to better understand their paracrine action, which may lead to novel therapeutic strategies in veterinary medicine., (© 2019 Humenik et al.)