Wound healing is a dynamic and tightly orchestrated process in which mechanical cues play essential roles in regulating cellular behavior, matrix remodeling, and tissue regeneration. Photocrosslinkable biomaterials are versatile platforms to study these effects because their physical properties, such as stiffness, viscoelasticity, and porosity, can be precisely tuned in situ within the geometrically complex wound environment. Here, we outline methods to investigate mechanobiological regulation of wound healing using photocrosslinkable biomaterials, with a focus on gelatin and polyethylene glycol as representative natural and synthetic biomaterials. We highlight the integration of these hydrogels with the murine excisional wound model to study wound healing dynamics and describe techniques to analyze healing, including histology, immunohistochemistry, functional assays, flow cytometry, and single-cell profiling. By linking biomaterial design to biological readouts, we provide a framework for dissecting how biomaterial mechanical cues guide tissue repair and regeneration and support the development of new strategies to enhance healing outcomes.
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