Introducing multiple physical cues to control cell behaviors effectively is considered as a promising strategy in developing bioactive wound dressings. Silk nanofiber-based cryogels are developed to favor angiogenesis and tissue regeneration through tuning hydrated state, microporous structure, and mechanical property, but remained a challenge to endow with more physical cues. Here, β-sheet rich silk nanofibers are used to develop cryogels with nanopore structure. Through optimizing crosslinking time and exposing the reactive group inside the nanofibers, the crosslinking reaction is improved to induce stable cryogel formation. Besides the hydrated state and macroporous structure, the nanopore structure formed on the macroporous walls, providing hierarchical microstructures to improve cell migration. Both in vitro and in vivo results reveal quicker cell migration inside the cryogels, which then accelerates angiogenesis and wound healing. The mechanical properties can further regulate to match with skin regeneration. The wound healing study in vivo reveals lower inflammatory factor secretion in the wounds treated with softer cryogels with nanopores, which then resulted in the best angiogenesis and wound healing with less scar. Therefore, the porous cryogels with multiple physical cues can be fabricated with silk nanofibers to control cell behaviors and tissue regeneration, providing a promising approach for designing bioactive wound dressings.