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西罗莫司处理制备新型小口径异种移植血管的研究
Experimental study of a novel bioengineered small-caliber vascular graft incorporating sirolimus

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DOI:
作者:
王学宁
Xue-Ning Wang
作者单位:
山西省心血管病医院
Department of Cardiovascular Surgery, Shanxi cardiovascular Hospital
关键词:
西罗莫司；内膜增生；小口径；肝素；脱细胞
Sirolimus, Intimal hyperplasia, Small-caliber, Heparin, Decellularization
摘要:
目的 借鉴西罗莫司药物洗脱冠状动脉支架的原理和技术，将经脱细胞和肝素处理的小口径异种移植血管进一步结合西罗莫司，观察术后移植血管内膜增生的情况，从而进一步改进小口径异种移植血管的性能。方法 取新鲜犬颈动脉，经脱细胞喝肝素结合处理后分为两组：A组（对照组，脱细胞+肝素）和B组（西罗莫司组，脱细胞+肝素+西罗莫司处理）。兔20只，将A、B两组血管同时分别植入同一只兔左、右双侧颈动脉。术后1、2、4、8周采血用HPLC法检测B组中西罗莫司的含量。术后3个月超声检测移植血管通畅情况，术后3和6个月分别随机处死10只兔，取双侧血管样本经免疫组织化学及HE染色观察血管的重塑及内膜增生情况。结果 B组血管术后1至8周均可检测到西罗莫司。术后3个月超声显示，A组部分血管斑块形成（5/20），造成管腔狭窄，B组斑块形成较少（2/20）（P<0.05），无明显管腔狭窄。HE染色显示，两组部分血管管腔内血栓形成，未形成血栓的两组血管管壁经免疫组织化学检测可见新生内皮细胞和平滑肌细胞。术后3和6个月，近、远端吻合口附近A组内膜增生比B组显著（P<0.05）。结论 进一步结合西罗莫司可以有效减轻内膜增生，提高这种脱细胞肝素处理的小口径异种移植血管的性能，有助于提高远期通畅率，使之可能作为小口径移植血管新的来源。
Objective The lessons from the principle and technology of sirolimus-eluting Coronary Stent were drawn and the effect of intimal hyperplasia were observed, The aim of this study was to develop a new small-caliber decellularized and heparinized vascular xenograft through sirolimus treatment. Methods Canine common carotid arteries were chosed as xenografts through decellularization and heparin treament. The xenografts with (n=20, group A) and without (n=20, group B) sirolimus coating were implanted in rabbits' left and right carotid artery respectively as bypass grafts. The content of sirolimus was measured by HPLC at 1, 2, 4, and 8 week in Group B, respectively. Graft patency were checked by Duplex ultrasonography at 3 months after implantation. Ten rabbits were euthanized randomly respectively at 3 and 6 months and bilateral grafts were explanted. Tissue samples were observed by HE staining, immunohischemical staining and SEM were performed to observe the vascular remodeling. HE staining and micrograph analysis system were used for valuating the intima hyperplasia (IH) of bilateral grafts. Results Sirolimus was found in vascular wall of Group B during the 8 weeks. At 3 months, checked by Duplex Doppler, there was a little plaque formation and vessel stenosis was found in some blood vessels of Group A (5/20). There was also a little plaque formation in Group B (2/20, P<0.05) with no significant vessel stenosis. HE staining showed thrombosis in some grafts in both groups. Dense α-smooth muscle actin staining of grafts without thrombosis suggested that cells populating the media were smooth muscle cells (SMCs), and Factor Ⅷ staining of cells covering the luminal surface of these grafts confirmed that they were endothelial cells (ECs) at 3 months. The intima thickness levels at both anastomatic stomas of Group A significantly was more serious than Group B（P<0.05）. Conclusion Sirolimus treatment can reduce intimal hyperplasia, better the capability, and help to improve the long-term patency of the vascular xenograft. The new xenograft can be hoped to become a new source of small-caliber vascular inplatation.

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