The aim of this study was to evaluate the healing of class III furcation defects following transplantation of autogenous periosteal cells combined with beta-tricalcium phosphate (beta-TCP). Periosteal cells obtained from Beagle dogs' periosteum explant cultures, were inoculated onto the surface of beta-TCP. Class III furcation defects were created in the mandibular premolars. Three experimental groups were used to test the defects' healing: group A, beta-TCP seeded with periosteal cells were transplanted into the defects; group B, beta-TCP alone was used for defect filling; and group C, the defect was without filling materials. Twelve weeks post surgery, the tissue samples were collected for histology, immunohistology and X-ray examination. It was found that both the length of newly formed periodontal ligament and the area of newly formed alveolar bone in group A, were significantly increased compared with both group B and C. Furthermore, both the proportion of newly formed periodontal ligament and newly formed alveolar bone in group A were much higher than those of group B and C. The quantity of cementum and its percentage in the defects (group A) were also significantly higher than those of group C. These results indicate that autogenous periosteal cells combined with beta-TCP application can improve periodontal tissue regeneration in class III furcation defects.