Eumelanin pigment release from photo-crosslinkable methacrylated gelatin-based cryogels: Exploring the physicochemical properties and antioxidant efficacy in wound healing

Managing wounds in certain phases of the healing process still represents a big challenge. The oxidative stress, caused by reactive oxygen species (ROS), is one of the hallmarks controlling the wound healing-related process. Multifunctional biomaterials with excellent biocompatibility, tuneable properties, and easy functionalization, may allow realizing suitable three-dimensional (3D) and extracellular matrix (ECM)-mimicking structures, to efficiently control ROS levels. This might be a promising strategy for healing severe wounds. Herein, photo-crosslinkable methacrylated gelatin (GelMA)-based spongy-like cryogels (from 5 to 20 % w/v) incorporating Eumelanin from Black Soldier Flies (BSF-Eumel, 0.5 and 1.0 mg/mL), a pigment endowed with marked antioxidant properties, were developed. GelMA-based cryogels were fabricated by an easily handled and scalable cryogelation process followed by ultraviolet (UV) photo-crosslinking. BSF-Eumel sub-micrometer particles were embedded into GelMA-based cryogels by passive permeation of the solution within the polymeric network. BSF-Eumel addition resulted in more hydrophilic and porous structures, exhibiting a good stability and a prolonged release within 14 days. Furthermore, GelMA/BSF-Eumel cryogels exhibited good antioxidant activity, confirmed by a powerful quenching effect on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (59 % at 1.0 mg/mL of BSF-Eumel). Moreover, GelMA/BSF-Eumel cryogels at the highest GelMA concentrations (10 and 20 % w/v) accelerated human dermal fibroblasts-adult (HDF-a) migration, promoting wound closure within 24 h. They also proved to mitigate oxidative stress, modulating the production of ROS levels and preventing superoxide dismutase (SOD) activity inhibition in HDFs stimulated by lipopolysaccharide (LPS), owing to the release of BSF-Eumel. Such remarkable outcomes make GelMA/BSF-Eumel cryogels a promising antioxidant platform for wound healing.