以纤维蛋白凝胶为支架构建组织工程胃新膀胱的研究

目的探讨以纤维蛋白凝胶为支架，通过组织工程技术以胃．重建膀胱的可行性。方法从实验动物获取膀胱黏膜，体外培养并扩增移行上皮细胞。分别通过纤维蛋白凝胶（实验组）和keratinocyte-SFM（对照组）将单个细胞悬液移植于带血管蒂的去黏膜胃片，将胃片缝成囊状（新膀胱），固定于腹壁。于术后2、4、6、8周取胃片进行组织学及免疫组化分析。结果实验组术后随时间延长新膀胱有不同程度挛缩；HE染色提示胃囊腔面有上皮样细胞覆盖，2周时细胞覆盖率约50％，4—8周时约70％。这些细胞分布不均匀，分层不明显；上皮样细胞AE1／AE3染色和CK7染色均阳性。对照组各标本均严重挛缩，腔隙消失，无上皮覆盖。结论将体外培养扩增的尿路上皮细胞通过纤维蛋白凝胶移植至去黏膜胃片，可以使去黏膜胃片移行上皮化，从而重建膀胱。

Study on the construction of gastric bladder using fibrin glue as the tissue engineering scaffold

Objective To explore the feasibility of constructing gastric bladder using fibrin glue as the tissue engineering scaffold. Methods Urothelial cells were isolated and propagated in vitro from open bladder biopsies taken from 17 pigs. The single cells were suspended in fibrin glue(experimental group) and keratinocyte-SFM (control group) respectively, and then were transplanted onto free patch of demucosalized stomach tissue based on the right gastroepiploic artery. We then sew the patch into pockets (artificial bladder) and fixed them onto the abdomen wall, retook the artificial bladder for histologic and immunohistochemical analysis at different postoperative interval (2, 4, 6 and 8 weeks). Results In the trial group, macroscopically, the demucosalized stomach pockets shrank in different degree. Histological analysis showed that the surfaces of the artificial bladder were covered by a layer of epithelial-like cells, which covered the surfaces unevenly and stratified unclearly. Immunohistochemical analysis showed that the covering epithelial-like cells were positively stained with CK7. In the control group, the artificial bladders shrank and adhered so gravely that there were no existing lacuna. Conclusion It is available to transplant the transitional cells cultured in vitro onto the demucosalized patch of stomach so as to reconstruct functional gastric baldder.