膀胱粘膜下层无细胞基质的研制

Study on preparation of bladder submucosa accellular matrix

目的探讨膀胱粘膜下层无细胞基质的制作与评价。方法自膀胱解剖出膀胱粘膜下层,置于PBS、0.5％SDS、双蒸水中反复洗涤去除细胞,必要时加用胰蛋白酶消化。获得的细胞外基质采用HE、免疫组化、扫描电镜评价去细胞效果,细胞植入无细胞基质观察细胞能否粘附于该材料。结果HE、免疫组化、扫描电镜均未见制备的材料中存在细胞及细胞碎片,体外细胞植入试验见细胞可粘附于材料并增殖。结论采用本制备方法可以得到膀胱粘膜下层无细胞基质,有望为临床提供尿道下裂生物替代材料。

Objective In recent years the application of extracellular matrix as a replacement of intact tissue for reconstitution has been the subject of intense study. Much has been learnt about the important role of bioscaffold in tissue and organ morphogenesis and for the repair of several tissues. The aim of this study was to explore the preparation of bladder submucosa acellular matrix and to evaluate its properties, efforts that might lead to the effective use as a template for urinary tract repair. Methods The bladder submucosa graft was derived from porcine bladder by microdissection. Muscular tissues should be removed so as to avoid remnants and at the same time the intactness of submucoas should be maintained. The isolated graft was then thoroughly washed in PBS, 0. 5% SDS, dH2O; trypsin digestion(0.1% trypsin:0. 02% EDTA = 1 : 1)was used at 37 ℃ for 2 hours before dH2O wash if there were any muscular remnants left. HE staining, immunochemical staining, scanning electronic microscope and cell seeding test in-vitro were carried out to evaluate the extracellular matrix. Results The cells can adhere to bladder submucosa acellular matrix and proliferate in it. Cells or remaining cellular elements were not seen at HE staining, immunochemical staining and scanning electronic microscope.Conclusions The results showed that bladder submucosa acellular matrix can be obtained by this methods and the acellular matrix may provide alternative biomaterial for hy-pospadias.