犬股动脉置换壳聚糖-硫酸化丝素蛋白人工血管:内皮层及血管层的生成

Canine femoral artery displacement with the chitosan-sulfated silk fibroin artificial blood vessel: the formation of vascular endothelial cells, vascular smooth muscle cells and fibroblasts

背景:前期实验制备了壳聚糖-硫酸化丝素蛋白人工血管。目的:通过犬股动脉置换壳聚糖-硫酸化丝素蛋白人工血管,探讨组织工程血管的功效性能。方法:将8只比格犬随机分为2组,实验组于股动脉进行壳聚糖-硫酸化丝素蛋白人工血管置换;正常对照组不进行任何处理。置换后6个月,检测凝血四项后,采用苏木精-伊红染色观察血管结构,免疫组织化学观察血管结构形成,电镜观察血管内皮层及血管层生成情况。结果与结论:1苏木精-伊红染色:实验组血管形成了内皮细胞层、平滑肌层和成纤维细胞层3层主要血管组织,在组织形态上与正常对照组血管无显著差别;2免疫组织化学染色:实验组FactorⅧ染色可见组织边缘一深色带染色即为内皮层形成;á-SMA染色可见较多棕色颗粒沉淀,为血管平滑肌形成;Vimentin染色亦可见较多棕色颗粒沉淀,提示人工血管也形成了成纤维细胞层。该3层组织与正常对照组血管无明显差异;3电镜观察:实验组人工血管内表面及切面与正常对照组血管相似,切面也形成了3层组织,但排列不及正常血管整齐;4凝血四项:两组凝血酶原时间、活化部分凝血活酶时间、凝血酶时间、纤维蛋白原无差异;5结果表明:壳聚糖-硫酸化丝素蛋白人工血管具有较好的生物相容性及功效性能。

BACKGROUND： The chitosan-sulfated silk fibroin artificial blood vessel was obtained in our previous study. OBJECTIVE： To investigate the biocompatibility and function of the chitosan-sulfated silk fibroin artificial blood vessel replacement into the canine femoral artery. METHODS： Eight Beagles were randomly allotted to two groups, and chitosan-sulfated silk fibroin artificial blood vessel was implanted into the femoral artery （experimental group）, but animalsin control group received no intervention. Six months after implantation, the coagulation function was detected, the vascular formation, including endothelial, smooth muscle and fibroblast layers were observed through hematoxylin-eosin staining under electron microscope. RESULTS AND CONCLUSION： （1） Hematoxylin-eosin staining showed that vascular endothelial cells, vascular smooth muscle cells and fibroblasts were found in the experimental group, which were similar with the normal vascular structures （2） Immunohistochemistry： in the experimental group, there was a deeply stained band at the tissue edege through Factor VIII staining, suggesting the formation of the endothelium; there were abundant brown particles precipitated in a-SMA staining, suggesting the vascular smooth muscle cell formation; Vimentin staining showed various brown particle precipitations indicating the fibroblast formation, and all were close to the normal vascular structures. （3） Electron microscope observed that the inner surface and section of the artificial vessel were similar with the normal one, and the three-layer structures formed in the section arranged irregularly. （4） There were no differences in the prothrombin time, activated partial thromboplastin time, thrombin time and fibrinogen between groups. To conclude, these results suggest that the chitosan-sulfated silk fibroin artificial blood vessel holds good biocompatibility and performances.