The purpose of this study was to analyze histomorphometrically the influence of the ratio of particulate autogenous bone (AB) graft/platelet-rich plasma (PRP) on bone healing in surgically created critical-size defects (CSD) in rat calvaria. Fifty rats were divided into five groups: Group C (control), Group AB, Group AB/PRP-50, Group AB/PRP-100, and Group AB/PRP-150. A 5-mm diameter critical-size defect was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In Group AB, the defect was filled with 0.01 mL of AB graft. In Groups AB/PRP-50, AB/PRP-100, and AB/PRP-150, the defects were filled with 0.01 mL of AB graft combined with 50, 100, and 150 mL of PRP, respectively. All animals were euthanized at 30 days postoperative. Histomorphometry, using image analysis software, and histology analyses were performed. New Bone Area (NBA) and the remaining bone graft particles area (RPA) were calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for analysis. No defect completely regenerated with bone. Group AB/PRP-50 (41.78 AE 13.48%) had a significantly greater NBA than Groups C (19.29 AE 5.11%), AB (27.43 AE 10.90%) or AB/PRP-150 (19.17 AE 8.45%) (p < 0.05). No significant differences were observed between groups AB/PRP-50 and AB/ PRP-100 or among groups AB, AB/PRP-100, and AB/PRP-150 with regard to NBA (p > 0.05). Group AB/PRP-150 (31.59 AE 3.22%) had a significantly greater RPA than Groups AB (19.09 AE 5.21%), AB/PRP-50 (17.33 AE 4.43%), and AB/PRP-100 (19.72 AE 3.62%) (p < 0.001). No significant differences were observed among groups AB, AB/PRP-50, and AB/PRP-100 with regard to RPA (p > 0.05). The ratio AB graft/PRP influences bone healing in surgically created CSD in rat calvaria. ß