This study aimed to investigate the bone healing capacity of using chitosan-gelatin sponge as bone substitute material in rabbit calvarial bone defects and to compare with autogenous bone grafting. Six New Zealand white rabbits were included in the study. Two identical 8 mm diameter cranial bone defects were prepared in each rabbit and grafting was done with autogenous bone from defect preparation and chitosan-gelatin sponge. After 12 weeks, bone defects of the two groups were evaluated qualitatively and quantitatively by means of radiography (step-wedge calibration, imaging densitometry) and histology (histomorphometry, digital imaging analysis). Results showed that chitosan-gelatin sponge promoted bone healing in part by its osteoconductive property. Up to 40-45% of areas of new bone formation were found in the specimens from chitosan-gelatin sponge group. There was statistical difference in both radiographic optical density and histomorphometric percentage bone area when comparing between the two groups (p < .05). To conclude, the use of only chitosan-gelatin sponge could not replace autogenous bone graft with regard to bone healing capacity. However, further studies of other modifications of chitosan-gelatin sponge grafting are indicated.