摘要
目的将通过分离培养的表皮细胞与脱细胞膀胱黏膜下筋膜复合,修复缺损的尿道,观察表皮细胞表型在尿道环境中的转归。方法采用同种异体(家兔)的膀胱,经显微分离和脱细胞液处理,制成无细胞的生物支架(并经随机切片证实脱细胞效果),选择兔的小块包皮组织,消化收集分离出的表皮细胞,经过增殖、传代培养植入生物支架中,并加入Brdu标记物,将其卷成管状,回植入自体人工造成的尿道缺损,术后1、2、6及12个月分别测定治疗效果(每组各处死3只家兔/批),观察指标:尿道造影、大体外形、修复段尿道黏膜的HE染色、免疫组化和荧光标记等。结果术后实验对象伤口愈合正常,修复尿道的大体形态和尿道造影显示排尿通畅,无尿瘘和狭窄发生;HE染色和免疫组化显示,术后1月—6月,修复段尿道黏膜层次由单一向复层结构转变;而术后12月修复段尿道黏膜已基本显示为类似移行上皮细胞结构;术后角蛋白染色阳性;Brdu标记在术后1月清晰显示植入上皮细胞层存在,随后逐渐稀少。术后6及12月尿道黏膜结构中未见显影。结论作为种子细胞,表皮细胞可与胶原筋膜结合运用于尿道修复,修复效果良好,并随着时间延长向尿道原有细胞结构转变。
Objective To investigate the transformation of epidermal cells from foreskin to reconstruct engineered anterior urethra with acellular collagen matrix. Methods Acellular collagen matrices were generated from allogeneic rabbit bladder submucosa. In 12 rabbits, autologous foreskin epidermal cells were isolated, in vitro expanded and seeded （inoculation） onto the tubular acellular collagen matrix. In 12 male rabbits, urethral mucous defect was created and urethroplasty was performed with tubular acellular collagen matrix seeded with epidermal cells （n=12）. Urethrography was performed at 1, 2, 6 and 12 months postoperatively. In addition, Urethras grafts were harvested and analyzed grossly and histologically. Results Urethrography and gross views showed a patency of repaired urethral defect with the maintenance of a wide urethral caliber and without sign of strictures at any time points. Histology showed the graft seeded with epidermal cell had formed single layers structure by 1 month, which demonstrated the characteristic presence of a epidermal cells layer with a normal submucosa structure. At 2, 6 month postoperatively, histology showed multi-layers epidermal cells with vessel abundance in submucosa structure. Evidence margin between graft epidermal cells and epithelium cell lays of hostage were at 6 month postoperatively. Histology showed transitional cell lays with graft urethra and without evidence margin between graft and hostage urethra at 12 month postoperatively. Conclusion As seed cells, epidermal cells seeded to acellular collagen matrix can reconstruct the urethra defect and transform to transitional cells structure finally.
出处
《中国男科学杂志》
CAS
CSCD
2008年第3期1-5,共5页
Chinese Journal of Andrology
基金
2005年度上海市教委科研课题基金<表皮细胞作为种子细胞构建尿道的实验研究>(05BZ19)
2003年度上海市卫生局科研课题基金<植入上皮细胞的胶原筋膜在尿道修复中的运用>(034082)
关键词
尿道疾病
组织工程
膀胱
urethral diseases
tissue engineering
