小肠黏膜下层与内皮细胞复合构建组织工程血管

Construction of tissue engineered blood vessels by combining small intestinal submu-cosa with endothelial cells

目的探讨以小肠黏膜下层为材料制作组织工程血管支架的可行性,并对其复合内皮细胞进行研究,从而为制备组织工程血管提供初步准备。方法以猪空肠为原材料,利用机械方法制备小肠黏膜下层,去除抗原性后对小肠黏膜下层进行免疫荧光染色,以观察其所含层粘连蛋白和纤维连接蛋白。以猪大隐静脉为原料分离培养并扩增大隐静脉内皮细胞。小肠黏膜下层与内皮细胞复合培养,并通过Hoechst33258染色、FDA/PI染色观察内皮细胞黏附和生长情况,从而评估小肠黏膜下层与内皮细胞的生物相容性。结果小肠黏膜下层含有较多层粘连蛋白和纤维连接蛋白,内皮细胞成功接种于小肠黏膜下层,并且存活数量较多,生长状态良好。结论小肠黏膜下层具有较好的生物相容性,可促进内皮细胞的黏附与生长,具有构建组织工程血管的能力,可行性较高。

Objective To investigate the construction feasibility of tissue engineered intravascular stent by applying small in-testinal submucosa(SIS)as materials and then study its composite endothelial cells seeded on SIS,thus which provides preliminarily preparation for tissue engineered blood vessels.Methods The small intestinal mucosa was prepared by mechanical methods with pig jejunum as raw material,immuno fluorescence staining of SIS was performed after removing the antigenicity to observe the expression of laminin and fibronectin.The endothelial cells of porcine great saphenous vein were isolated,cultured and amplified.The epithelial cells were seeded on SIS,then the endothelial cell adhesion and growth were observed by Hoechst 33258 staining and FDA/PI stain-ing,thereby assessing the biocompatibility of small intestinal submucosal cells with endothelial cells.Results The SIS contains more laminin and fibronectin,the endothelial cells were successfully inoculated on SIS,many of which survived and grew in good condition.Conclusion The SIS showed good biocompatibility and therefore promoted endothelial cell adhesion and growth after being seeded on SIS,which provided potential to build tissue engineered blood vessels and high feasibility.