Crosslinking methods for skeletal muscle extracellular matrix-derived hydrogels

Volumetric muscle loss is a pathological condition characterized by an extensive loss of muscle tissue that overwhelms the regenerative capacity of skeletal muscle, resulting in lifelong functional deficits. Current treatments for severe muscle loss are limited, including autologous muscle flaps, muscle transposition, and synthetic patches, all of which have significant drawbacks, such as donor site morbidity, limited availability of donor tissue and graft failure, resulting in minimal muscle regeneration. To avoid additional surgeries, decellularized extracellular matrix (dECM)-derived hydrogels represent a promising biological alternative for tissue replacement. Although they can promote a favorable regenerative microenvironment, they possess poor mechanical properties. To enhance these, crosslinking strategies can be employed. The aim of this thesis is to evaluate the effect of two different crosslinking methods for dECM-derived hydrogel reinforcement: the addition of genipin, a natural chemical crosslinker, and the application of mechanical stimulation using a custom-designed bioreactor to induce a physiological strain. The effect of the crosslinkers is evaluated using collagen quantification assay, enzymatic degradation analysis, scanning electron microscopy, rheometric properties, fluorescence recovery after photobleaching test and finite element analysis.