ABSTRACT
Background: Bone grafting is a preferred treatment option for the healing of large diaphyseal bone defects and is useful in the management of nonunion, delayed union, and tumor resection. Aims: To investigate a decellularization protocol of bovine cancellous bone for xenogenic implantation in radial bone defects in rabbits. Methods: Bovine bone scaffolds fabricated with various decellularization protocols viz phosphate buffer saline (PBS), 1% sodium dodecyl sulfate (SDS), and rapid freeze and thaw technique. The manufactured scaffolds were characterized by biomechanical testing, histological staining, and scanning electron microscopy. A 10 mm rabbit radius bone defect was repaired with autograft and SDS treated and rapid freeze and thaw in groups A, B, and C respectively. Healing was evaluated by radiography and histopathology at 0, 30, 60, and 90 days. The grafts were also checked for host tissue reaction and incorporation into the defect. Results: The freeze and thaw group showed complete elimination of all cellular nuclei, regular arrangement of collagen fiber, and no significant difference in tensile strength compared to 1% SDS treated and native groups. The in vivo radiographic and histopathological study showed that the rapid freeze and thaw group had complete bridging of the bone gap defect with new bone formation and they were immunologically less reactive compared to group B. Conclusion: The in vitro and in vivo evaluation of the grafts suggested that freeze and thaw technique was most superior to all other techniques for effective decellularization and augmentation of bone healing with better integration of the graft into the host.
ABSTRACT
AIM: The aim of this study was to generate composite bone graft and investigate the rabbit fetal osteoblasts adhesion, proliferation and penetration on acellular matrices of cancellous bone. MATERIALS AND METHODS: Acellular cancellous bone was prepared and developed as in the previous study with little modification. These matrices were decellularized by rapid freeze and thaw cycle. To remove the cell debris, they were then treated with hydrogen peroxide (3%) and ethanol to remove antigenic cellular and nuclear materials from the scaffold. Primary osteoblast cells were harvested from 20 to 22 days old rabbit fetal long and calvarial bone. These cells were cultured and characterized using a specific marker. The third passaged fetal osteoblast cells were then seeded on the scaffold and incubated for 14 days. The growth pattern of the cells was observed. Scanning electron microscope and hematoxylin and eosin staining were used to investigate cells proliferation. RESULTS: The cells were found to be growing well on the surface of the scaffold and were also present in good numbers with the matrix filopodial extensions upto inside of the core of the tissue. CONCLUSION: Thus, a viable composite scaffold of bone could be developed which has a great potential in the field of bone tissue engineering.