Bioresorbable polymeric membranes for guided bone regeneration (GBR) were fabricated using the three-dimensional printing technique. Membranes made of polylactic-co-glycolic acid (PLGA), which consist of lactic acid (LA) and glycolic acid in ratios of 10:90 (group A) and 70:30 (group B), were compared. Their physical characteristics including architecture, surface wettability, mechanical properties, and degradability were compared in vitro, and their biocompatibilities were compared in vitro and in vivo. The results demonstrated that the membranes of group B had mechanical strength and could support the proliferation of fibroblasts and osteoblasts significantly better than those of group A (p< 0.05). The degradation rate in Group B was significantly lower than that in Group A, but they significantly produced less acidic environment (p< 0.05). In vivo, the membranes of group B were compared with the commercially available collagen membranes (group C). The amount of newly formed bone of rat’s calvarial defects covered with the membranes of group C was stable after week 2, whereas that of group B increased over time. At week 8, the new bone volumes in group B were greater than those in group C (p >0.05). In conclusion, the physical and biological properties of the PLGA membrane (LA:GA, 70:30) were suitable for GBR.