原位与体内构建小鼠组织工程阴道的实验研究  

Comparison between Vaginal Epithelial Cells Seeded and Unseeded Porcine Acellular Dermal Matrix for Vagina Reconstruction in Mice

作　　者：张明乐[1] 黄向华[1] 李雅钗[1] 张琳[1] 李亚楠[1] 朱志洁[1] 王振海[1] 

机构地区：[1]河北医科大学第二医院,河北石家庄050000

出　　处：《实用妇产科杂志》2013年第4期277-281,共5页Journal of Practical Obstetrics and Gynecology

基　　金：河北省医学适用技术跟踪项目(编号:GL2011-36)

摘　　要：目的:比较原位组织构建与体内构建两种方式在小鼠体内进行阴道重建的效果。方法:切除昆明小鼠阴道后采用酶消化法获得阴道上皮细胞。A组(12只)植入猪脱细胞真皮基质(PADM)行原位组织构建;B组(12只)植入自体阴道上皮细胞与纤维蛋白-PADM移植复合物行体内构建。于移植后2周、4周、8周、12周取出重建的阴道组织进行大体观察,并分别进行伊红-美蓝染色(HE)、Van Gieso(VG)染色和广谱角蛋白AE1/AE3免疫组化染色,以动态观察重建阴道组织生长情况及PADM降解情况。重建术后12周用透射电镜观察重建阴道组织超微结构。结果:①A组在移植后2周可见2～3层细胞的上皮形成;移植后4周细胞层数增加,但细胞排列无极向;移植后8周、12周上皮层结构接近正常小鼠阴道上皮层结构。B组移植后2周上皮结构即接近正常小鼠阴道上皮结构;移植后4周、8周、12周上皮层结构基本同移植后2周,但细胞形态更加成熟,极向更为明显。移植后12周A组PADM部分区域结构不完整,B组PADM仍保持基本轮廓。②AE1/AE3免疫组化染色证实了重建阴道的上皮化。③B组透射电镜观察可见上皮细胞之间存在桥粒连接,上皮细胞与基底膜之间有半桥粒连接。上皮层下见成纤维细胞、无髓神经纤维与毛细血管。结论:利用PADM进行原位组织构建与体内构建在实验动物体内均能够完成阴道重建。在技术成熟的条件下,体内构建法比原位组织构建法更适合于进行阴道重建。Objective：To compare the effect of vagina reconstruction between with vaginal epithelial cells seeded and with unseeded porcine acellular dermal matrix （PADM） in an animal model. Methods： Epithelialcells were isolated by enzymatic digestion after vagina excision in Kunming mice. Vagina reconstruction with vaginal epithelial cells unseeded and seeded PADM were performed in group A（12 mice） and group B（12 mice） ,correspondingly. At week 2,4,8 and 12 after implantation,the animals were sacrificed and the neova- ginas were prepared for hematoxylin-eosin （HE） staining,Van Gieson （VG） staining and epithelial broad spectrum of monoclonal antibodies of AE1/AE3 immunohistochemical staining to evaluate the status of vari- ous layer growth of vagina. The ultrastructure of neovagina was observed by transmission electron micro- scope at week 12 after implantation. Results： ①In group A,only 2 -3 layers of epitheliums were observed under the light microscopy at week 2 after implantation. At week 4,epithelization in mucosa of neovaginal ca- nal was intensified to 4- 5 layers, but the epithelial cells were disarranged. After 8 - 12 weeks,the differences between the neovagina and the native vagina were harldy noted. In group B,the neovagina had the same structure as the native vagina after 2 weeks. Within 4 -12 weeks the similar results could be observed, except that the cell shape became more mature and polarized. At week 12,degradation of PADM could be observed in group A,while the structure of PADM still remained complete. ②Immunohistochemical staining for cytoker- atins A1/A3 confirmed that vaginal epithelial cells was successful reconstructed. ③In group B,desmosomes, hemi desmosomes,fibroblasts, nerve fibre and capillary could be observed under transmission electron micro- scope. Conclusions..Vagina reconstruction with vaginal epithelial cells seeded and unseeded PADM are both feasible. With mature technology,vagina reconstruction with vaginal epithelial cells seeded is better than un- seeded P

关 键 词：阴道重建 组织工程 脱细胞真皮基质 纤维蛋白凝胶 阴道上皮细胞 

分 类 号：R711[医药卫生—妇产科学]