Nature derived biopolymers such as polysaccharides and collagen have attracted considerable attention in biomedical applications. Despite excellent biocompatibility and bioactivity, their poor mechanical properties could not meet the requirement for skin regeneration. In this study, cellulose nanocrystal (CNC) was incorporated into the calcium cross-linked sodium alginate/gelatin (SA/Ge) scaffold to reinforce its physicochemical properties. A novel sodium alginate/gelatin/cellulose nanocrystal (SA/Ge/CNC) scaffold was successfully prepared through electrostatic interaction of sodium alginate and gelatin, ionic cross-linking of calcium ions with sodium alginate, and incorporation of CNC. Afterwards, the SA/Ge and SA/Ge/CNC scaffolds were fully characterized and compared with scanning electron microscopy images, swelling behaviors, tensile strengths and contact angles. The involvement of CNC produces a hybrid SA/Ge/CNC scaffold with desired porous network, moderate swelling behavior, and superior mechanical strength (from 18 MPa to 45 MPa). Furthermore, in vitro cytotoxicity and cell growth assay using mouse embryonic fibroblast cells validated that SA/Ge/CNC scaffold was non-toxic and can prompt cell adhesion and proliferation. The in vivo skin regeneration experiments using the SA/Ge/CNC scaffold group showed an improved skin wound healing process with accelerated re-epithelialization, increased collagen deposition and faster extracellular matrix remodeling. Overall, the results suggested that the SA/Ge/CNC hybrid scaffold with enhanced mechanical performance and wound healing efficacy was a promising biomaterial for skin defect regeneration.