Background: Different treatment modalities have been developed to arrest the periodontal disease and to rebuild the lost periodontal tissues. Applied to periodontal biology, tissue engineering has the potential to improve the regeneration of lost tooth-supporting structures in a more predictable manner than conventional periodontal treatment.Aim of the study: The aim of the present study was to evaluate the potential of bone marrow mesenchymal stem cells (BMMSCs) expanded by tissue culture on periodontal regeneration of periodontal defects in rabbits. Materials and methods: BMMSCs were isolated from New-zealand rabbits and expanded in vitro. BMMSCs were evaluated for in vitro osteogenic differentiation and mineralized nodule formation, adipogenic differentiation, and Chondrogenic differentiation. Intra bony periodontal defects in the rabbits received a sterile collagen sponge alone and BMMSCs seeded on collagen sponge. Defects covered with gingival flaps only served as a control. All Animals were sacrificed 4 weeks postoperatively and specimens were evaluated by histological and morphometric analysis.Results: BMMSCs show significant periodontal regeneration potential in terms of bone, periodontal ligament (PDL), and cementum while collagen sponge alone was associated with limited formation of new bone, new cementum like tissue and new PDL in induced periodontal defects in rabbits. In addition, the control exhibited limited if any bone regeneration.Conclusion: The implantation of BMMSCs seeded on collagen sponge generated new bone, new cementum like tissue and new periodontal ligament while collagen sponge alone was associated with limited formation in induced periodontal defects in rabbits.