CAJ | 학술논문

目的探讨将骨髓间充质干细胞(BMSCs)和膀胱平滑肌细胞(SMCs)种植于膀胱脱细胞基质(BAM)上构建组织工程片状移植物(TESG),修复尿道缺损.方法制作兔BAM,体外培养扩增兔BMSCs和SMCs,将BMSCs和SMCs种植到兔BAM的两面,培养7d后制作成3.0×1.5cm2的TESG作为实验组,将未种植细胞的BAM构建TESG作为对照组.新西兰兔25只,其中实验组20只,分4组,每组5只,对照组5只,修复尿道缺损后分别于手术后2周、4周、8周取标本做HE染色,16周后对TESG进行免疫组化检测.结果脱细胞基质HE染色及扫描电镜未见细胞生长.实验组HE染色发现实验组尿路上皮层再生呈连续性;术后16周免疫组化可见TESG覆盖多层尿路上皮结构及规则的平滑肌束.对照组5只兔术后拔除尿管4周内均死亡,尸检见尿道明显狭窄,双侧肾脏重度积水,双测输尿管扩张,膀胱充盈明显.结论将BMSCs和SMCs种植于BAM上构建TESG,修复尿道缺损是可行的,BMSCs能促进尿路上皮再生.
목적 탐토장골수간충질간세포(BMSCs)화방광평활기세포(SMCs)충식우방광탈세포기질(BAM)상구건조직공정편상이식물(TESG),수복뇨도결손.방법 제작토BAM,체외배양확증토BMSCs화SMCs,장BMSCs화SMCs충식도토BAM적량면,배양7d후제작성3.0×1.5cm2적TESG작위실험조,장미충식세포적BAM구건TESG작위대조조.신서란토25지,기중실험조20지,분4조,매조5지,대조조5지,수복뇨도결손후분별우수술후2주、4주、8주취표본주HE염색,16주후대TESG진행면역조화검측.결과 탈세포기질HE염색급소묘전경미견세포생장.실험조HE염색발현실험조뇨로상피층재생정련속성;술후16주면역조화가견TESG복개다층뇨로상피결구급규칙적평활기속.대조조5지토술후발제뇨관4주내균사망,시검견뇨도명현협착,쌍측신장중도적수,쌍측수뇨관확장,방광충영명현.결론 장BMSCs화SMCs충식우BAM상구건TESG,수복뇨도결손시가행적,BMSCs능촉진뇨로상피재생.
Objectives To study the feasibility? of a tissue-engineered sheet graft for urethral reconstruction using bone marrow mesenchymal stem cells (BMSCs) and smooth muscle cells (SMCs) seeded bladder acellular matrix (BAM).Methods BAM of rabbits were made in vitro.Rabbit BMSCs and SMCs were isolated and expanded in vitro before seeding into BAM as the experimental group,whereas BAM-only was the control group.In experimental group,20 New Zealand rabbits,divided into 4 groups and each of five,and five other were the control group.Specimens were obtained at 2 week,4 weeks,8 weeks after surgery respectively and were stained with haematoxylin and eosin.Epithelium and smooth muscle of TESG was detected with Immunohistochemistry 16 weeks postoperatively.Results HE staining and scanning electron microscopy revealed no residual cells adhering to BAM.Histologic examination revealed the regeneration of the urothelium on the inner lumen of TESG was consistent.Immunohistochemistry showed anti-cytokeratin AE1/AF3 antibody and anti-uroplakin Ⅲa antibody positive,confirming that there were mature and functional epithelial cells on the lumen of TESG,and organized smooth muscle bundles were present.Five rabbits in the control group died within 4 weeks postoperatively.Autopsy showed their urethras to be almost completely blocked or urethral strictures.Conclusions It is feasible to construct TESG using BMSCs and BAM for urinary diversion in rabbits and BMSCs could promote the regeneration of urothelium.