适用于犬输尿管脱细胞基质制备的灌注系统

目的探讨应用灌注系统制备输尿管脱细胞基质的可行性方案。方法利用输尿管本身存在的管腔结构，通过灌注系统制 备输尿管脱细胞基质。根据化学洗涤剂的不同特性设置3 个不同的灌注组，单纯SDS 组、单纯TritonX-100 组以及联合组 （TritonX-100+SDS）。利用HE染色，DAPI染色，DNA定量，琼脂糖凝胶电泳评价输尿管脱细胞基质中细胞核的残留情况。采 用Masson’s 3色染色、Alcian Blue染色、胶原含量测定、GAG含量测定、扫描电镜和毒性检测等手段评价制备输尿管脱细胞基质 的三维结构和生物活性成分。结果HE染色和DAPI染色显示联合组制备的输尿管脱细胞基质中没有明显的核物质残留， DNA定量和凝胶电泳证实了这一结果。Masson’s 3色染色、Alcian Blue染色、胶原含量测定和GAG含量测定显示联合组保留 了输尿管脱细胞基质三维胶原结构和生物活性成分。扫描电镜观察联合组制备的输尿管脱细胞基质表面存在大量的孔隙结 构。毒性测定证实联合组制备的输尿管脱细胞基质无毒性。结论本研究灌注系统可用于输尿管脱细胞基质的制备，筛选出一 套比较理想的脱细胞方案，制备的输尿管脱细胞基质可用于输尿管组织工程再造。

A perfusion system for preparing canine ureteral acellular matrix

Objective To explore the feasibility of preparing ureteral acellular matrix (UAM) using perfusion systems. Methods Using the luminal structure of the ureter, the UAM was prepared by perfusing canine ureter with SDS, TritonX-100, or both. The residual nuclei in the UAM were evaluated using HE staining, DAPI staining, DNA quantification, and agarose gel electrophoresis. The three-dimensional ultrastructure and the bioactive components were evaluated by Masson’s trichrome staining, Alcian Blue staining, collagen quantification, GAG quantification, scanning electron microscopy (SEM), and toxicity detection. Results HE staining and DAPI staining showed the absence of obvious nuclear materials in the combined group, which was further confirmed by DNA quantification and agarose gel electrophoresis. Masson’s trichrome staining, Alcian Blue staining, collagen quantification and GAG quantification all verified that the ultrastructure and the bioactive components were well preserved in the combined group. SEM showed a large amount of porous structure on the surface of the UAM prepared by combined perfusion, and toxicity assay confirmed that the prepared UAM was nontoxic. Conclusion Perfusion of canine ureter with SDS and TritonX-100 is feasible to prepare UAM for ureteral reconstruction.