Background: To examine the effects of the addition of autologous platelet-rich plasma (PRP) into bilayer poly(lactide-co-glycolide) (PLGA) scaffolds on the reconstruction of osteochondral defects in a rabbit model. Material/Methods: Porous PLGA scaffolds were prepared in a bilayered manner to reflect the structure of chondral and subchondral bone. Bone defects, measuring 4 mm in diameter and 4 mm in thickness, were created in both knee joints in 18 healthy New Zealand white rabbits, aged between 120-180 days old. Rabbits were randomly divided into three groups: rabbits with bone defects implanted with bilayer PLGA scaffolds (PLGA group) (N=6); or with bilayer PLGA and autologous PRP (PLGA/PRP group) (N=6); and the untreated group (control group) (N=6). The gross morphology, histology, and immunohistochemistry for the expression of collagen type II and aggrecan were observed at 12 weeks after surgery and compared using a scoring system. Micro-computed tomography (CT) imaging and relative expression of specific genes were also assessed. Results: The platelet concentrations in the PRP samples were found to be 4.9 times greater than that of whole blood samples. The total score on gross appearance and histology was greatest in the PLGA/PRP group, as was the expression of collagen II and aggrecan of the neo-tissue. Micro-CT imaging showed that more subchondral bone was formed in the PLGA/PRP group. Conclusions: Bilayer PLGA scaffolds loaded with autologous PRP improve the reconstruction of osteochondral defects in the rabbit model.