The purpose of this study was to evaluate new bone formation and healing process in rat calvarial bone defects using BioMesh(R) membrane and DFDB. Forty eight rats divided equally into 4 groups of 1 control group and 3 experimental groups. Standardized transosseous circular calvarial defects (8 mm in diameter) were made midparietally. In the control group, the defect was only covered with the soft tissue flap. In the experimental group 1, it was filled with DFDB only, in the experimental group 2, it was covered BioMesh(R) membrane only, and in the experimental group 3, it was filled DFDB and covered with membrane. At the postoperative 1, 2, 4, 8 weeks, rats were sacrificed and histologic and histomorphometric analysis were performed. These results were as follows. In histomorphometric analysis, It showed the greatest amount of new bone formation through experimental in the experimental group 3 (P < 0.001). The amount of new bone formation at the central portion of the defect was greater in the experimental group 3 than experimental group 2. BioMesh(R) membrane began to resorb at 1 week and resorbed almost completely at 8 weeks after operation. The collapse of membrane into the defect was observed through the experimental periods in the experimental group 2. In the area of collapsed membrane, new bone formation was restricted. These results suggest that maintenance of some space for new bone to grow is required in the use of BioMesh(R) membrane alone in the defect. It is also thought that use of the membrane may promote new bone growth in DFDB graft.