The ability of collagen-based resorbable barrier membranes to stabilize microenvironments is a current clinical concern for guided bone regeneration (GBR) technology. Decellularized dermal membranes are a research hotspot now, which can retain the natural three-dimensional extracellular matrix (ECM) structure and provide a positive microenvironment for cell adhesion and proliferation, thereby promoting tissue healing and regeneration. However, imperfect decellularization methods often result in weaker performance than collagen membranes in clinical practice (e.g., Bio-Gide membrane). Hence, we propose a composite decellularization technique combining chemical and biological enzymatic methods, supplemented by physical techniques, to obtain a high-performance decellularized dermal membrane for GBR. The prepared decellularized dermal membrane effectively maintained the original 3D microstructure and natural components (collagen and elastin) of the ECM, exhibiting good physical properties and biocompatibility. Decellularized dermal membranes can promote fibroblast adhesion and growth on the surface while effectively preventing fibroblasts from invading the injured site. Moreover, the decellularized dermal membrane showed the potential to induce osteogenesis and inhibit osteoclast formation. In vivo cranial defect experiments demonstrated the superior GBR performance of the decellularized dermal membrane compared to commercial Bio-Gide membrane.