Rapid and functional skin wound healing remains a major clinical challenge, yet current biomaterial-based therapies largely function as passive scaffolds or delivery vehicles. Despite their widespread clinical use, whether biomaterials possess intrinsic bioactivities capable of actively instructing endogenous regenerative programs remains poorly understood. Here, independent of bulk structural or mechanical cues and exogenous biomolecules, we demonstrate that silk fibroin (SF), a clinically approved biomaterial, intrinsically programs keratinocytes into a Ccl27⁺ migratory-paracrine pro-regenerative state that coordinates epidermal-dermal crosstalk to drive regenerative wound healing. Using integrated scRNA-seq, proteomics, and keratinocyte-specific genetic perturbation, we show that SF engages the integrin receptor ITGA2 on keratinocytes to activate the PI3K-Akt pathway, relieving p53-mediated repression of Ccl27. Elevated Ccl27 endows keratinocytes with enhanced migratory capacity to facilitate re-epithelialization, while Ccl27 secreted from SF-activated keratinocytes simultaneously acts in a paracrine manner to promote fibroblast proliferation and dermal remodeling, collectively enabling efficient wound closure and appendage reconstruction. Keratinocyte-specific conditional knockdown of Ccl27 largely abrogates the pro-regenerative effects of SF in vivo. SF elicits only minimal immune activation during wound healing, preserving an immunocompatible regenerative microenvironment. These findings uncover an intrinsic bioactivity of SF that actively programs a pro-regenerative keratinocyte state, establishing a paradigm in which biomaterials function as instructive cues rather than passive scaffolds to drive functional tissue repair.