Study of mechanical stimulation as a conditioning strategy for dECM hydrogel patches in muscle defect therapy

This study proposes the development of engineered patches based on hydrogels derived from decellularized extracellular matrix (dECM) of porcine origin, aimed at the treatment of muscle defects, to overcome the limited regenerative capacity of skeletal muscle tissue. The main objective of the thesis was to investigate whether the application of mechanical stimuli during the patch culture phase would lead to the formation of tissue with characteristics similar to the physiological one. To evaluate the regenerative potential, a mixture of human muscle cells (85%) and fibroblasts (15%) was embedded in a dECM-derived hydrogel. Mechanical stimulation was then applied both during the hydrogel crosslinking phase and during long-term culture, using a customizer bioreactor to mimic the biomechanical conditions of skeletal muscle. Cultures were analyzed for up to 10 days under both static and mechanically stimulated conditions. Samples were analyzed using 2D and 3D immunofluorescence analyses, scanning electron microscopy, DNA amount quantification, and enzymatic degradation assays.