静电纺丝素蛋白材料复合尿路上皮细胞修复尿道的实验研究  

Experimental study of electrospun silk fibroin matrices seeded with urothelial cells for urethral reconstruction

作　　者：谢敏凯[1] 宋鲁杰[1] 汪继洪[1] 范苏娜[2] 张耀鹏[2] 徐月敏[1] 

机构地区：[1]上海交通大学附属第六人民医院泌尿外科,200233 [2]东华大学纤维材料改性国家重点实验室

出　　处：《中华泌尿外科杂志》2014年第8期629-634,共6页Chinese Journal of Urology

基　　金：国家自然科学基金（81170641）

摘　　要：目的 评估拉伸处理后的静电纺丝素蛋白材料与尿路上皮细胞复合在尿道重建中的应用效果.方法 2012年1-9月,制备新型的经拉伸处理的静电纺丝素蛋白材料.使用扫描电镜检测和孔隙率检测方法评估丝素材料结构.使用9只比格犬作为实验动物,其中实验组6只、对照组3只.从实验组动物中获得、培养、扩增其尿路上皮细胞,并种植于材料上,培养1周,获得组织工程移植物,并经过HE染色剂扫描电镜检测.实验动物被剥离尿道黏膜3 cm×1 cm,实验组应用组织工程移植物进行修复,对照组未进行修复.术后1、2及6个月后,进行尿道造影及组织学检测.结果 扫描电镜及孔隙率检测显示经拉伸处理的静电纺丝素蛋白材料为三维多孔结构.尿路上皮细胞在丝素材料上生长良好,在材料表层多层生长,且细胞能渗透入材料内部.在动物实验中,实验组动物未表现排尿困难,尿道造影示尿道未见明显狭窄,组织学检测显示术后1个月1～2层上皮细胞覆盖修复部位,上皮细胞逐渐生长,在术后6个月形成典型的尿路上皮细胞结构.对照组动物出现排尿困难,尿道造影示尿道管腔狭窄,组织学检查显示上皮细胞生长缓慢,术后1个月未形成或仅见断续上皮层,并可见较严重的炎性反应,术后2个月形成2～3层细胞结构,术后6个月尚未形成完整的尿路上皮层.结论 拉伸处理后的静电纺丝素蛋白材料有良好的生物相容性.复合尿路上皮细胞的静电纺丝素蛋白材料或可成为潜在的尿道重建材料.Objective To investigate the feasibility of urethral reconstruction by using stretched electrospun silk fibroin matrices.Methods Stretched electrospun silk fibroin matrix was prepared,and the structure of the material was assessed by electron microscopy.Canine urothelial cells were isolated,expanded and seeded onto the material for 1 week to obtain a tissue-engineered graft.The tissue-engineered graft was assessed using HE staining and electron microscopy scanning.A dorsal urethral mucosa defect was created in 9 female beagle dogs.In the experimental group,tissue-engineered mucosa was used to repair urethral mucosa defects in 6 dogs.No substitute was used in the 3 dogs of the control group.Retrograde urethrography was performed at 1,2 and 6 months after grafting.The urethral grafts were analyzed grossly and histologically.Results Electron microscopy scanning revealed that the material had a 3 dimensional porous structure.Urothelial cells grew on the material and showed good biocompatibility with the stretched silk fibroin matrices.Canines implanted with tissue-engineered mucosa voided without difficulty.Retrograde urethrography revealed no signs of stricture,and histological staining showed gradual epithelial cell development and stratified epithelial layers at 1,2 and 6 months.The canines in the control group showed difficulty in voiding.Retrograde urethrography showed urethral stricture,and histological staining showed that no or only one layer of epithelial cells developed.A severe inflammatory reaction was also observed in the control group.Conclusion Stretched electrospun silk fibroin matrices have good biocompatibility with urothelial cells,and could be a potential material for urethral reconstruction.

关 键 词：尿道 组织工程 丝素蛋白 生物材料 移行上皮细胞 

分 类 号：R318.08[医药卫生—生物医学工程]