Cyclosporine-impregnated allograft bone sterilized with low-temperature plasma

Abstract

Deep-freezing, freeze-drying and gamma (γ)-irradiation have deleterious effects on bone healing and mechanical properties of allograft bones. We tried preparing bone allografts using cyclosporine plus low-temperature-plasma sterilization. To explore the feasibility of this method of preparation, segmental defects in the right radii of rabbits were repaired with cyclosporine-impregnated allograft bones (CABs) sterilized with low-temperature-plasma (in the study group) and deep-frozen/freeze-dried irradiated allograft bones (D/FIABs) (in the control group). X-ray and quantitative histological analysis, peripheral blood T lymphocyte subset analysis and CD₂₅ molecule immunohistochemistry stain, the four-point bending test and safety evaluations were respectively conducted to compare bone-healing, immunosuppression, mechanical properties and safety between the two groups. X-ray scores were higher in the study group than those in the control (p = 0.032). There were significant differences in new bone areas at most repairs in both groups (p < 0.05). There were no significant differences in the percentages of CD₄(+) T, CD₈(+) T, ratios of CD₄(+) T:CD₈(+) T or serum concentrations of GPT/Cr in both groups (p > 0.05). At 16 weeks postoperatively, the density of CD₂₅ molecules in the control group was higher than that in the study group. The ultimate loading in the study group was significantly higher than that in the control (p = 0.048). Bone marrow stromal cells (BMSCs) grew thickly around and on the surface of a cyclosporine-impregnated allograft. Livers and kidneys in the study and control groups remained histologically normal at 7 days postoperatively. These results indicate that the CAB might be a better material than the D/FIAB in terms of bone healing, preservation of mechanical properties and immunosuppression without severe side-effects.