SUMMARY:Recent studies revealed multipotent properties of fat tissue isolated mesenchymal stem cells. These cells are successfully used as therapeutic factor for many locomotive disorders, being even more effective than stem cells from bone marrow. Isolated and cultured, AD-MSCs were observed, photographed and measured to compare cells from two different species.

A horse suffering from an undetected hoof bone fracture was diagnosed three weeks after injury. The formation of callus tissue was detected at the fracture site. Standard orthopaedic screw application was augmented by a novel method, a combination of stem cells and plasma components. For experimental therapy, fat tissue and blood samples were collected from the patient to isolate stem cells and plasma proteins. The obtained and characterised mesenchymal stem cell population was applied to the wound area, together with an implant prepared from plasma, wrapped over the orthopaedic screw. Additionally, cells with implant were differentiated in vitro into bone tissue, to evaluate if cells could successfully produce extracellular matrix in such material. Three weeks after application, the hoof was significantly regenerated, and after three months -the bone was completely rebuilt. The in vitro experiment also gave positive results, with completely differentiated cells after three weeks. Our data show that enriching the standard orthopaedic material with mesenchymal stem cells adds therapeutic value to the treatment of refractory bone fractures.

Background and Objectives: AD-MSCs (adipose-derived mesenchymal stem cells), as an easy obtainable population of multipotent cells, have been successfully applied in many diseases in animals. Having very similar properties and morphology to these collected from bone marrow, are very attractive object for regenerative medicine. Methods and Results: Ovine stem cells applications were considered to serve as suitable animal model for large animals locomotive system disorders therapy. In this research, sheep fat was collected from four different regions of the body. AD-MSC were isolated and cultured for morphological, morphometrical and immunophenotypical analysis. On the basis of conducted study, cells isolated from topographically different part of the body of given animal were almost identical. Morphologically and immunologically, they are very similar to other species. Moreover, application of light and electron microscopy, as well as immunolabeling and measurement assays allowed for detail morphological and biofunctional analysis of ovine adipose-derived mesenchymal stem cells in the culture. Conclusions: Ovine subcutaneous fat from many regions of the body is rich source of stem cells, which could be used in locomotive system disorders experimental therapy on sheep models.

Enteropathogenic E. coli (EPEC) is a human pathogen using type III secretion system for delivery of proteins directly into the human host. The system contains a single ATPase, EscN, which is essential for uncoupling of proteins from their complexes with chaperones before the delivery. The structure of EscN ATPase (PDB code: 2obm) was used to screen computationally for small molecule inhibitors blocking its active site. Two lead candidates were examined but only one, Compound 54, was selected for further optimization. After extended QSAR optimization, two derivatives were found to be competitive inhibitors of EscN capable of blocking ATPase activity with a K below 50 µM. One candidate, WEN05-03, with a K=16±2 µM, was also minimally toxic to mammalian cells as determined by other assays. In the cell infection model of HeLa cells with EPEC, Compound WEN05-03 completely blocked actin cluster formation at 100 µM concentration, when analyzed by confocal microscopy. The second best inhibitor of EscN ATPase activity was WEN04-34 with a K=46±2 µM. However, the compound was highly toxic to the BALB/3T3 cell line. In summary, the work identifies a compound blocking bacterial ATPase in its active site without causing cellular toxicity to the host cells. It is the first report showing feasibility of using bacterial virulence system ATPase as a target for safe, non-toxic compounds and offering a proof-of-concept for non-antibiotic alternatives.

BACKGROUND: Great potential of cellular therapies has generated extensive research in the field of cells harvesting and culturing. Transplantation of cell cultures has been used in a variety of therapeutic programs but in many cases it appeared that biomaterial scaffold or sheath would enhance cells regenerative potential. OBJECTIVE: Hydrogels composed of different proportions sodium and calcium alginates, were undertaken to evaluate their influence on mesenchymal stem cells and olfactory bulb-derived glial cells cultures. Additionally, these biomaterials were also enriched with fibrin protein. METHODS:The structure of materials was visualized by means of scanning electron microscopy. After seeding with cellshydrogels were observed with inverted and fluorescence microscope. Cell's morphology, behavior and phenotype were analyzed in investigated materials by means of light, fluorescence and scanning electron microscopes. Also, viability assay was performed with Alamar Blue cytotoxic test. RESULTS: Our observations showed that basic alginate hydrogels had significant influence on both cell types. Materials maintained cells alive, which is desired attribute, however none of them kept cells in normal, flat form. Alginates with significant calcium component kept cells alive for longer period of culture. CONCLUSIONS: Addition of fibrin protein resulted in material's biocompatibility properties improvement, by creation of adhesion surface, which helps cells to keep proper morphology and behavior. Our findings suggest that addition of fibrin protein to alginate hydrogels improves them as cell carriers for regenerative medicine applications.