The aim of this study was to evaluate the bone repair using autogenous periosteum-derived cells (PDC) and bovine anorganic apatite and collagen (hA-COl). PDC from Wistar rats (n=10) were seeded on hA-COl discs and subjected to osteoinduction during 6 days. Critical-size defects in rat calvarias were treated with blood clot (G1), autogenous bone (G2), hA-COl (G3) and hA-COl combined with PDC (G4) (n=40), and then analyzed 1 and 3 months after surgeries. radiographic analysis exhibited no significant temporal change. G1 and G2 had discrete new marginal bone, but the radiopacity of graft materials in G2, G3 and G4 impaired the detection of osteogenesis. At 3 months, histopathological analysis showed the presence of ossification islets in G1, which was more evident in G2, homogeneous new bone around hA-COl in G3 and heterogeneous new bone around hA-COl in G4 in addition to moderate presence of foreign body cells in G3 and G4. histomorphometric analysis showed no change in the volume density of xenograft (p>0.05) and bone volume density in G2 was twice greater than in G1 and G4 after 3 months (p<0.05), but similar to G3. The PDC did not increase bone formation in vivo, although the biomaterial alone showed biocompatibility and osteoconduction capacity.Key Words: critical size defects, periosteum-derived cells, xenograft, hydroxyapatite, collagen.to obtain the osteogenic cells. Also, PDC show greater proliferation, which decreases the time of cell culture, contamination risks and experiment costs. It could be a favorable alternative for dentists as a regenerative therapy for alveolar bone to treat periodontitis and dental implant dehicenses, due to the easy handling, expansion and re-implantation of cells (2).Cell culture is done upon scaffolds to simulate the 3D structure of bone tissue. Demineralized and acelullar bovine xenograft matrices are predominantly composed