The Therapeutic Effect of Autogenic Adipose Derived Stem Cells Combined with Autogenic Platelet Rich Plasma in Tendons Disorders in Horses in vitro and in vivo Research

Naturally, occurring tendons injuries including superficial digital flexor tendinopathy are the most frequent musculoskeletal disorders in. performance horses. Conventional methods of treatment with non steroidal and steroidal anti-inflammatory medicaments in majority of cases lead to scar formation, reducing the quality and efficiency of tissue regeneration. Novel approach is aimed to use cells naturally present in an organism as regeneration enhancing factor. In conducted research, the intralesional injections of autologous adipose derived stem cells combined with autologous platelet concentrate therapeutic potential was investigated in horses with 8-12 weeks duration superficial digital flexor injury with severe scaring. Collected by clinical examinations data showed positive effects of autologous, adipose-derived mesenchymal stem cells combined with autologous platelet rich plasma injections on regeneration processes in the course of superficial flexor tendon injures in horses. On the basis of ultrasound examination, it was proved that the quality of healed tissue was significantly higher in experimental group, comparing to control group. Obtained results confirmed the beneficial pro-regeneration effects of stem cells/platelet concentrate combined injections. The obtained data may also serve as valuable source of information about morphology and behaviour of fat stem cells in culture or platelets appearance

Design of small molecule inhibitors of type III secretion system ATPase EscN from enteropathogenic Escherichia coli

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 Ki below 50 µM. One candidate, WEN05-03, with a Ki=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 Ki=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