Projects

The role of tissue non-specific alkaline phosphatase (TNAP) in osteoarthritis (OA) has not been fully clarified. High expression of TNAP as ectonucleotidase is associated with ATP degradation and production of adenosine, which can again trigger and support inflammatory calcifying conditions, but also decrease mineralization of the subchondral bone in OA. Thus, we anticipate that TNAP beyond its well-known contribution to mineralization, participates in profiling of cellular metabolism. To verify the assumption that TNAP controls metabolism of cells from OA patients, we attempt to apply genome editing (GE) techniques to silence and knockout Alpl gene in OA patient-derived mesenchymal stromal cells. We will perform Alpl gene silencing (commercially available siRNA) and CRISPR/Cas9 technique to establish OA patient-derived GE cell lines (clones) with Alpl gene silencing/knockout for investigation of metabolic roles of TNAP in OA patients.

Large bone defects present a common clinical problem where autologous bone grafting is the current gold standard. Autologous bone grafts harvested by the Reamer-Irrigator-Aspirator® (RIA) technique are considered advantageous in terms of larger quantities of graft material and yielding two fractions – a solid graft usually used for the transplantation and a liquid fraction often referred to as a waste material. This highly translational project aims to address the regenerative potential of RIA liquid fraction through in vitro studies of its secretory and cellular content as well as by testing pre-conditioning strategies to improve RIA grafts regenerative potential. Also, the project will provide evidence for the new application possibility of bone substitute material PerOssal® (Osartis GmbH) in combination with RIA-graft material. In vitro experiments will be translated into preclinical and clinical trials in Serbia to optimize currently available therapies. In parallel, proof-of-concept experiments with 3D printing materials will be accomplished with the industrial partner from Serbia 3D Republika d.o.o.