Objective To investigate the effect of local administration of recombinant adenovirus of human adiponectin (hAPN) Ad-hAPN-EGFP on tibial defect repair of SD rats. Methods Tibial defect (2 mm) models of 36 SD rats (72 sides) were randomly divided into three groups (A, B, and C; n=24). The three groups were injected with Ad-hAPN-EGFP, Ad-EGFP, and normal saline, respectively, during operation and the day after operation. A week after the operation, the expression of hAPN and osteogenesis-related factors were detected by real-time polymerase chain reaction. Three weeks after operation, the tibias were examined by micro-computed tomography, hematoxylin-eosin staining, and Masson staining to evaluate the restoration of bone defects. Results 1) The hAPN expression was detected in group A but not in groups B and C. Osteogenesis-related factors expression of group A was significantly higher than that of the other groups (P<0.05). 2) Osteogenesis (including bone mineral density, relative bone volume, trabecular number and trabecular thickness) in group A were more evident than those in groups B and C (P<0.05). No significant differences were found between groups B and C (P>0.05). Conclusion Local administration of recombinant adenovirus Ad-hAPN-EGFP may be an effective strategy to improve the restoration of bone defects in vivo.