Objectives Different forms of allogenic dentine have been studied for their potential use as bone substitutes. We report a new method for processing bovine dentine that results in a sterile bioactive material for repair and regeneration of bone. Methods Extracted bovine dentine was processed mechanically and chemically with inorganic and organic solvents, and sterilised. The in vitro biocompatibility on human gingival fibroblasts was assessed by the Alamar Blue assay and the in vivo biocompatibility evaluated by implantation of the processed dentine into rats' femurs. Results The dentine showed excellent biocompatibility in vitro, stimulated formation of new bone and was completely incorporated into the new bone in vivo. Significance Processed bovine dentine has the potential to be used as a suitable substitute in bone repair and regeneration.