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The purpose of this study was to determine the effect of calcium sulfate (CS) on cementum deposition and osseous healing after periradicular surgery. The root canals of 24 mandibular premolars in four 2-yr-old beagle dogs were endodontically treated, followed 2 weeks later by periradicular surgery. Mineral trioxide aggregate (MTA) was used as root–end-filling material. The right or left side was assigned at random to receive CS alpha-hemihydrate or no material in the osteotomy sites before closure. The animals were killed after 4 months. Hard-tissue healing was analyzed histomorphometrically. All samples displayed evidence of cementum deposition adjacent to the root-end fillings and bone regeneration in the osteotomy sites. The data was analyzed using the Mann-Whitney U test, comparing the scores for cementum and osseous healing of the two groups at significance level of alpha = 0.05. The results indicated that placement of CS in osteotomy sites after periradicular surgery does not significantly affect periradicular healing.