Development of composite bubaline cancellous bone xenografts by seeding guinea pig fetal osteoblasts

: Xenografts are considered as an alternative strategy to restore a critical sized bone defect. The present study was conducted to standardize the technique for seeding and cryopreservation of decellularized bubaline cancellous bone with Guinea pig fetal osteoblast for the development of a composite bone xenograft. The composite bone grafts were prepared by seeding the expanded osteoblasts on the acellular bubaline cancellous bone matrix. The foetal osteoblast seeded scaffold, acellular bone scaffold, and native bovine bone were analyzed using scanning electron microscopy (SEM) and histological examination. Composite bone xenograft was cryopreserved in 10% glycerol at - 80°C for three months and evaluated for post-thaw viability. Findings from the in vitro study suggested that the seeding of acellular scaffold was adequate, and osteoblasts had good adhesion and proliferation inside pores of acellular bone matrix. Histologically the integrity of collagen matrix was best preserved in the acellular group as compared to the freshly seeded and cryopreserved scaffold. The osteoblast seeded decellularized bubaline cancellous bone xenografts can be preserved for three months with adequate cell viability in the post-thaw evaluation at – 80°C in sterile tubes containing 10% glycerol as the cryoprotectant.