| Objective:The graft reconstruction of large skeletal defects after malignant bone tumors resection and trauma is an challenging problem which has been troubled orthopedic industry.Although there are many ways to repair the defects, such as arthrodesis,prosthesis implantation, autologous joints graft and allogeneic joints transplantation, but a variety of methods have advantages and disadvantages. Currently, allogeneic joints transplantation is widely used in clinical, but the clinical application is greatly limited by the immunity issue. This study attempts to further investigate body’s immune system reactions after cryopreserved allograft of joint from the immunological aspects, which provides a theoretical basis for the effective application of clinical transplantation of allogeneic joints.Methods:In this study, osteoarticular allograft were Implemented between inbred rats with different histocompatibility, refering experimental animal models which was designed by Yaremchuk et al. 30 Wistar rats were used as donors, providing 60 allogeneic knees, 90 Sprague-Dawley(SD) rats as recipients, including 80 as receptors, and the other 10 as syngeneic transplant donors.80 SD rats were in turn divided into A(fresh syngeneic transplant group), B(fresh allograft group), C(cryopreserved allograft group) and D(with cyclosporine A group of fresh allogeneic transplantation) into four groups and each group has 20 rats.Operator cutted off all donors’ s knees at knees’ s each 1cm, eliminated the soft tissue around the joints, used saline to irrigate bone canal,and then transplanted donors’ s knees to the same position abdominal subcutaneous of all recipient.Among them, the allogeneic joint which would graft to the group C rats is subject to two steps freezing treatment before transplantation,the postoperative D group rats were continuous application of cyclosporine A for two weeks.Recipient peripheral bloods were collected on week 0, 1, 2,3,4,6,8,10 and 12 post transplantation for detecting the levels of serum lymphocyte subpopulations andcytokines IL-2,IFN-r, IL-10 and IL-4 with the help of ELISA and flow cytometry.Results:1)CD4+/ CD8+and CD25+in four groups did not change significantly in the first two weeks.group B and C then gradually increased from the second week, and reached its peak in the eighth week. Group D began to increase from the fourth week,and reached maximum in the tenth week, while group A did not change significantly in the entire measurement period. Between the third week and twelfth week, group B index was significantly higher than group A(p <0.01), and higher than the C, D groups(p <0.05), CD4+/ CD8+and CD25+values changes ware statistically significant; the measured indicators in group C were higher than those in group A(p <0.05), the changes ware statistically significant. The measured indicators in group D rised from fourth week,and higher than group A(p<0.05), the difference was statistically significant,but compared with group C, the difference was not statistically significant,(p> 0.05).2)Cytokines IL-2, IFN-r, IL-10 and IL-4 did not change significantly in the first two weeks after surgery, then cytokines in group B and C began to gradually increase from the second week, and reached its peak in the eighth week.Group D began to increase from the fourth week, and reached a maximum in the tenth week, while there was no significant change in group A among the entire measured period. Between the third week and twelfth week, IL-2 and IFN-r of group B was significantly higher than group A(p <0.01),and higher than group C and D, the difference was statistically significant.The measured result of group C and group D was higher than the group A(p <0.05),but there was no statistically significant(p> 0.05) between group C and D. IL-10 and IL-4 of group C and group D was significantly higher than group A and B(p <0.01),there was a statistically significant difference;while we compared two groups of metrics between group C and group D, there was no difference(p> 0.05) and the difference was not statistically significant; group B measured by IL-10 and IL-4 of group B was higher than the group A, the difference was statistically significant,(p <0.05).Conclusion:1.Cryopreserved allogeneic joints transplantation can induce certain degree of immune rejection.2.Cryopreservation can reduce immune rejection of allogeneic joints transplantation.3.It is necessary to do histological typing before allogeneic joints transplantation.4.Immunosuppressants can delay immune rejection of allogeneic joints transplantation. |
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