Background. Bone healing is a complex and dynamic process that represents a well-orchestrated series of biological events of cellular recruitment, proliferation, and differentiation. The use of medicinal plants in bone healing has attracted increasing interest because of their lower side effects. Punica granatum seed oil (PSO) contains high levels of phenolic compounds, promotes osteoblast function, and plays an important role in bone remodeling. A gelatin sponge (Spongostan) is a hemostatic agent that is extensively applied as scaffolds in engineering and as drug carriers in the medical field. This study aimed to evaluate the effectiveness of PSO for bone healing enhancement. Twenty adult male New Zealand rabbits, weighing an average of 1.5–2 kg, were used in this study. Three intrabony holes were created in the tibiae of each animal, which were filled with a gelatin sponge (GS group) and combined gelatin sponge and PSO (GS/PSO group). Holes without material application were designated as the control group (C group). The animals were sacrificed at the healing duration (2–4 weeks) to prepare bone specimens for histological and histomorphometric analyses. Results. Histological findings indicated that the bone defects in the GS/PSO group showed more bone formation, mineralization, and maturation compared with the C and GS groups. Multiple group differences for bone cells showed a highly significant difference among all groups in the 2- and 4-week healing periods except for the C/GS and GS/GS/PSO groups at 4-weeks duration. Furthermore, highly significant results were obtained between both durations regarding the trabecular area, trabecular number, and bone marrow area. Conclusion. The study revealed that the combined application of GS and PSO was more effective in enhancing bone regeneration and accelerating bone healing compared with the other groups.