摘要
背景:由于创伤、先天畸形、肿瘤或手术继发的尿道缺损、狭窄、憩室等病理状况下,需要采用自体组织修复其缺损。目的:验证构建组织工程化尿道的可行性。设计、时间及地点:观察性实验,于2005-01/2006-01在中山大学林百欣医学研究中心完成。材料:雄性新西兰兔10只,体质量3.0～3.5kg,用于制备脱细胞尿道;4～6周龄BALB/cA-nude裸鼠20只,体质量18～20g,与细胞外基质材料复合体进行体内培育。方法:取新西兰兔完整尿道,脱细胞后形成细胞外基质材料,切取成体新西兰兔阴茎头约1/2大小,组织块法培养扩增兔平滑肌细胞;之后将尿道海绵体平滑肌细胞种植到脱细胞尿道基质上,将其植入裸鼠背部皮下进行培育。实验分为2组,实验组埋置细胞材料复合体,对照组埋置未接种细胞的单纯脱细胞尿道基质。主要观察指标:植入后1,2,4,6,8周取材,行大体和组织学及电镜观察细胞材料复合体的生长情况,免疫组织化学鉴定平滑肌细胞的表达。结果:①大体观察裸鼠背部切口愈合良好,饮食活动正常。植入后1,2周实验组脱细胞基质表面已有少量的组织膜形成;4周组织膜逐渐增厚,有小血管长入;6,8周脱细胞基质材料逐渐被新的细胞替代。②苏木精-伊红染色可见,随着时间延长,脱细胞材料逐渐被吸收,被各种细胞代替。其中平滑肌细胞最多,并可见大小不等的血管长入基质,部分有形成血窦的倾向;提示复合材料在裸鼠体内逐渐形成类似正常尿道的组织结构。③电镜观察从第4周有血管长入基质,可见较多的平滑肌细胞和内皮细胞及少量的成纤维细胞和巨噬细胞,并有部分平滑肌细胞凋亡。④α-平滑肌肌动蛋白抗体进行免疫组织化学染色后可见有平滑肌细胞生长。结论:采用组织工程技术可再造出类似于正常尿道的海绵体组织。
BACKGROUND： Under the pathological conditions of urethral defect, stricture and diverticulum induce by trauma, congenital malformation, tumor and operation, autogenous tissue are used for defect repair.
OBJECTIVE： To verify the feasibility of urethra constructed by tissue engineering. DESIGN, TIME AND SETTING： An observational experiment was performed at Linbaixin Medical Research Center, Sun Yat-sen University, from January 2005 to Janual-y 2006. MATERIALS： Ten New Zealand rabbits weighing 3.0-3.5 kg were used to prepare acellular urethras. Twenty BALB/cA-nude mice weighing 18-20 g, 4-6 weeks old, were cultured with extracellular matrix materials in vivo. METHODS： The whole urethra harvested from New Zealand rabbit prepared the extracelhilar matrix materials by acellular treatment. Half a glans harvested from adult New Zealand rabbit was used for smooth muscle cells proliferation by tissue explant. The corpus spongiosum smooth muscle cells were implanted in acellular matrix materials. Then, they were cultured by subcutaneous embedding in nude mice. There were two groups in the experiment： experimental and control. The complexs of corpus spongiosum smooth muscle ceils implanted in acellular matrix materials were embedded in the experimental group, while the simple acelhilar matrix materials were embedded in the control group.
MAIN OUTCOME MEASURES： After 1, 2, 4, 6, 8 weeks of embedment, the growth of the complexs were examined by gross, histologic and electron microscopic observation. The expression of smooth muscle cells were identified by immunohistochemistry method.
RESULTS： （1）Under the gross observation, incisions on the back of the nude mice healed well, the diets and activities of the nude mice were normal. In the experimental group, there were thin tissue membranes on the surface of the acellular matrix materials at 1-2 weeks after the embedment. The thickness of tissue membrane increased gradually and there were small blood vessels grown at 4 weeks. The new cells were instead o
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第50期9806-9810,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
