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Objectives: to evaluate the effect of milled teeth versus beta tricalcium phosphate bone graft material on new bone formation at the cell level using transmission electron microscope Materials and methods: Forty-five adult male albino rats, were divided into three main groups (n=15); control and two experimental groups. Under general anesthesia, 1.5 cm incision was done exposing the cranial bone. Then two noncritical size bone defects were created one in each parital sides of the rat calvarial bone. In Group I (control group): the bilateral calvarial bony defects were left to heal spontaneously. In Group II the bilateral calvarial bony defects completely filled with β-TCP. In Group III: the bilateral calvarial bony defects completely filled with tooth ash. Five rats from each group were sacrificed after 7, 15, and 30 days postoperatively. Results: tooth ash group: showed more new bone formation. Different types of cells appeared more active: multiple lymphocytes, fibroblast, osteoprogenitor cells, osteoblasts. Multinucleated osteoclasts with prominent vacuolated cytoplasm and ruffled border in Howship’s lacunae were clearly seen. At day 30, disappearing of the tooth ash and infiltration of different types of cells; osteoprogenitor cells, bone lining cells, and osteoblasts while β-TCP particles were still seen in the sections. Conclusion: Bone activity, formation and maturity were ahead in tooth ash filled calvarial bony defects. Tooth ash had a role in the stimulation of osteoblastic activity in matrix formation and its mineralization.