The ultimate goal of periodontal treatment is the regeneration of periodontal tissues which have been lost due to periodontal disease. Recently, many natural medicines have been studied for their potential of anti-bacterial, anti-inflammatory and regenerative effects in periodontal tissues. Safflower seeds have been traditionally used as a drug for treatment of fracture and blood stasis in oriental medicine. The objective of the present study is to examine the biologic effects of safflower seeds extract on bone formation and regeneration of rat calvarial defects. The calvarial defects were made with 8mm trephine bur and extract of safflower seeds were placed directly at these defects. 24 rats were divided into control and experimental groups, and each group was sacrificed at 1 week, 4 weeks and 8 weeks. To study a histopathology related to bone regeneration, Goldner's Masson Trichrome stain and histomorphologic measuring was done at each weeks. In the early phase of bone healing, less inflammatory infiltration and capillary proliferation was found in experimental group compared to control. Dense bony tissues and matured bone structures in defect areas were found in experimental groups. And area of new bone formation was significantly increased at 8 weeks in experimental group. These results indicate that direct local application of safflower seeds extract reduces the early inflammatory response and promotes the regeneration of new bone in calvarial defects of rats.