A novel micro-CT analysis for evaluating the regenerative potential of bone augmentation xenografts in rabbit calvarias

Guided Bone Regeneration is a common procedure, yet, as new grafting materials are being introduced into the market, a reliable evaluation method is required. Critical size defect in animal models provides an accurate simulation, followed by histological sections to evaluate the new bone formation. However, histology is destructive, two-dimensional and technique-sensitive. In this study we developed a novel volumetric Micro-CT analysis to quantify new bone formation characteristics. Eight adult female New Zealand white rabbits were subjected to calvarial critical-size defects. Four 8 mm in diameter circular defects were preformed in each animal, to allow random allocation of four treatment modalities. All calvarias were scanned using Micro-CT. Each defect was segmented into four equal parts: pristine bone, outer, middle, and inner. Amira software (v. 6.3, www.fei.com ) was used to calculate the new bone volume in each region and compare it to that of the pristine bone. All grafting materials demonstrated that new bone formation decreased as it moved inward. Only the inner region differed across grafting materials (p = 0.001). The new Micro-CT analysis allowed us to divide each defect into 3D regions providing better understanding of the bone formation process. Amongst the various advantages of the Micro-CT, it enables us to quantify the graft materials and the newly formed bone independently, and to describe the defect morphology in 3D (bi- vs. uni-cortical defects). Providing an insight into the inner region of the defect can better predict the regenerative potential of the bone augmentation graft material. Therefore, the suggested Micro-CT analysis is beneficial for further developing of clinical approaches.