玻璃化深低温法与去细胞光氧化技术制备同种异体小动脉体内性能比较

Comparison of in Vivo Performance of Small-diameterallogenic Arterioles Treated with Decellularization Associated with Photo-oxidation and Cryopreservation

目的:通过兔异体颈动脉旁路移植模型,对玻璃化深低温及去细胞光氧化交联处理的兔颈动脉进行体内性能比较,评价两种血管作为临床异体小血管替代物的可行性。方法:将36只SPF级纯种兔随机分成两组,分别予以玻璃化深低温处理的兔颈动脉及去细胞光氧化处理的兔颈动脉行异体颈动脉移植。实验动物分别饲养观察1周、2周和4周后,观察移植血管的通畅情况及内膜增生情况。结果:超声检查发现术后4周去细胞光氧化组的累计通畅率显著高于玻璃化深低温组(P<0.05)。HE染色显示玻璃化深低温组术后免疫反应表现比较严重。去细胞光氧化组术后内皮细胞则生长良好,炎性细胞浸润少。Ⅷ因子染色显示,术后4周玻璃化深低温处理组内皮细胞有不同程度的破坏;而去细胞结合光氧化组内皮生长良好。通过图像分析发现,移植后1周两组血管内/中膜比差异无统计秀而意义(P>0.05),术后2周及4周内膜两组血管内/中膜比差异有统计学意义(P<0.05)。结论:体内性能方面,去细胞光氧化技术制备同种异体小动脉生物稳定性、抗血栓性及内膜增生情况均要优于玻璃化深低温处理的异体兔颈动脉,但同样也存在瘤样扩张,内膜增生等不足。

Objective:To evaluate the feasibility of an alternative source of allogeneic arterioles in coronary artery bypass grafting,we build a model of rabbit allogeneic carotid bypass grafting and compared the in vivo performance of rabbit carotid arteries treated with decellularization associated with photooxidation and cryopreservation.Method:A total of 36 SPF purebred rabbits were randomly divided into two groups and separately given carotid bypass surgery with small-diameter allogenic arterioles treated with decellularization associated with photo-oxidation and cryopreservation.The experimental animals were reared for 1 week,2 weeks and 4 weeks.the hemodynamics and patency of graft vessels were observed through transvascular ultrasound and the intimal hyperplasia of the grafts were observed.Result:Vascular ultrasound examinations revealed that the cumulative patency rate of decellularization associated with photo-oxidation group was significantly higher than that of cryopreservation group(P<0.05).HE staining showed that the endothelial cells in the cryopreservation group had different degrees of shedding and hyperplasia,thrombosis in the lumen,and the immune response was much more severe.While the endothelial cells grow more well in the decellularization associated with photo-oxidation group with less thrombus formation and less inflammatory cell infiltration at the 4th week after surgery.Ⅷfactor staining showed that endothelial cells were disrupted and disorganized in the cryopreservation group while endothelial growth was much better in the ecellularization associated with photo-oxidation group 4 weeks after surgery.The image analysis system showed that there was no significant difference in intima/media ratio after the first week of bypass surgery(P>0.05),but significant difference began to occur between the two groups at 2 and 4 weeks postoperatively(P<0.05).Conclusion:In terms of in vivo performances,the biological stability,antithromboticity,and intimal hyperplasia of rabbit allograft in the decellularization associated with photooxidation group were all superior to those of cryopreservation group,but there are also tumor-like expansion,intimal hyperplasia and other deficiencies existing for us to modify.