Several techniques have been introduced to improve the pulp revascularization outcomes. The use of the tissue graft can create more practical tissue regeneration, provide vascular supply, and enhance tissue healing. The aim of the present study was to investigate the histologic and molecular outcomes of pulp revascularization with buccal fat autotransplantation. Fifty-six open apex roots from four dogs aged 4-6 months were randomly allocated to five groups of endodontic regeneration models: Group 1 (negative control, n = 4); Group 2 (control and without intervention, n = 4); Group 3 (blood clot, n = 16); Group 4 (buccal fat autotransplantation, n = 16); and Group 5 (blood clot plus buccal fat autotransplantation, n = 16). After 3 months, the extracted dog teeth were analyzed by histological and immunohistochemical techniques. Furthermore, real-time quantitative polymerase chain reactions were implemented to assess the gene expression profiles of dentin sialophosphoprotein (DSPP), dentin matrix protein (DMP), collagen I (COL1), and alkaline phosphatase (ALP) on regenerated tissue in the root canals. There were no significant differences in the severity of inflammation and necrosis between intervention groups. Immunohistochemical analysis showed significant differences among the study groups in expression level of extracellular glycoproteins such as fibronectin, laminin, and tenascin C. Group 5 showed an increase in the expression of DMP1 and COL1 genes. The expression of DSPP gene increased significantly in Group 4. The expression of ALP gene increased significantly in Group 3. Using this procedure may open new fields of research for regenerative endodontic procedure in which tissue autotransplant, particularly adipose tissue, may improve the outcomes of pulp revascularization.