Experimental Study Of The Osteolysis On Mice Prosthetic Loosening Model Restrained By Recombinant Adenovirus Ad-TNF-α-siRNA-BMP-2

Objective: To investigate the effects of gene therapy on mice prosthetic-loosening-modelby using recombinant adenovirus carried the sequence of TNF-α-siRNA-BMP-2.Methods:240female BABL/C mice were divided randomly into five groups (n=48):blank control group (A) used prosthetic-model, positive control group (B)、BMP-2controlgroup (C)、TNF-α-siRNA control group (D) and Combination gene control group (E) usedprosthetic–loosening-model. Two weeks later, each group were injected with the followingsolution40ul(PBS+24h later PBS)、(Ti solution+24h later PBS)、(Ti solution+24h laterAd-BMP-2)、(Ti solution+24h later Ad-TNF-ɑ-siRNA) and (Ti solution+24h laterAd-TNF-ɑ-siRNA-BMP-2), Biweekly repeated injections to the14week, normal fed for16weeks. Mouse were scanned by Micro-ct at12and16week,then tissue were harvested forstaining of HE、TRAP and immunohistochemical. Other tissues of mice were harvested at4、8、12and16week to get expression levels of TNF-α and BMP-2by ELISA test.Results: At same time, BMD and BVF of B、C、D、E、A group were increased in turn,the difference was statistically significant (P <0.05); limiting membrane were decreased andthe difference was statistically significant (P <0.05) between each group except D and E;BMD, BVF were reduced and limiting membrane were increased in group B and C whentime past, the difference was statistically significant (P <0.05).The TRAP staining show that osteoclasts of group D and E compared with group B andC have reduced significantly, the difference was statistically significant (P <0.05); there is nosignificant difference between group D and E、group B and C(P>0.05); osteoclasts were increased in group B and C when time past, the difference was statistically significant (P<0.05).Immunohistochemical staining showed that the expression levels of TNF-α was nosignificant difference between group D and E (P>0.05), it was lower than the group of B andC but higher than group A, and the difference was statistically significant (P <0.05).At same time，ELISA test showed that the expression of TNF-α in group D and E haveno difference (P>0.05); it was lower than Group B and C, but higher than in group A, and thedifference was statistically significant (P <0.05);the expression of TNF-α have no differencein group A when time past (P>0.05) and it was increased varyingly in the other group, thedifference was statistically significant (P <0.05).The expression of BMP-2in group C and Ehave no difference at same time(P>0.05); it was higher than Group A、B and D, the differencewas statistically significant (P <0.05); the expression of BMP-2was increased gradually ingroup C and E when time past (P<0.05) and there is no significant difference among the othergroup(P>0.05).Conclusion: Ad-TNF-ɑ-SiRNA-BMP-2could transfect and express effectively in thenormal cell, it could improve the efficiency of bone formation while reduce osteolysis, itcould be a efficient treatment of osteolysis;Ad-BMP-2can promote bone formation and havea compensatory role in the early osteolysis, but can not reduce the inflammatory reaction;Ad-TNF-α-siRNA can reduce the inflammatory reaction and osteolysis effectively, it could bea treatment of osteolysis.