Nonperforated polytetrafluoroethylene (PTFE) membranes are effectively utilized in guided bone regeneration (GBR) but may hinder cell migration due to limited interaction with the periosteum. This study compared bone regeneration using occlusive or perforated membranes combined with acellular collagen sponge (ACS) and recombinant human bone morphogenetic protein-2 (rhBMP-2) in a canine mandibular model. Male Beagle dogs (n = 3) received two mandibular defects each to compare ACS/rhBMP-2 with experimental (perforated group) and control (nonperforated group) membranes (n = 3 defects/group). Tissue healing was assessed histomorphologically, histomorphometrically, and through volumetric reconstruction using microcomputed tomography. The perforated group showed increased bone formation and reduced soft tissue formation compared to the nonperforated group. For the primary outcome, histomorphometric analysis revealed significantly greater total regenerated bone in the perforated group (67.08% ± 6.86%) than the nonperforated group (25.18% ± 22.44%) (P = .036). Perforated membranes had less soft tissue infiltration (32.91% ± 6.86%) than nonperforated membranes (74.82% ± 22.44%) (P = .036). The increased permeability of membranes in the perforated group potentially enabled periosteal precursor cells to have greater access to rhBMP-2. The availability may have accelerated their differentiation into mature bone-forming cells, contributing to the stimulation of new bone production relative to the nonperforated group.