The biological properties of morselised bone allograft treated with either a supercritical fluid process or low-dose (15 kGy) gamma irradiation were compared using radiological, histological and immunohistological techniques. The aims were to investigate any differences in the biological properties of supercritical fluid treated allograft and low-dose gamma irradiated allograft in-vivo. Rabbit allograft were cleaned of all soft tissue, cartilage and processed into ‘corticancellous crunch’ using a Noviomagus Bone Mill. Pooled samples were either gamma irradiated (15 kGy) or treated by NovaSterilis using super critical carbon dioxide. A well-reported tibial defect model in ten rabbits was used to examine the in vivo response of the different treatments at two and four weeks following surgery (n=5 per time point). Radiographic (x-ray, CT and micro CT), histology and immunohistochemistry was used to assess the in vivo response. Radiographic results revealed an initial response to the gamma-irradiated samples compared to SCF. Histology confirmed this reaction to be inflammatory in nature at two weeks that continued at four weeks for the gamma irradiated samples. In contrast, the SCF treated sample demonstrated new bone formation while the inflammatory reaction was muted compared to the gamma irradiated samples. Four week x-rays and histology confirmed new bone formation in both groups while the lack of significant inflammatory response in the SCF group was noted. Allograft sterilisation techniques do not result in the same initial response when evaluated in vivo. Removal of lipids and cellular debris following SCF treatment may influence the in vivo response. While both techniques can provide a sterile product, the in vivo response requires further investigation.
