天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (6): 584-587.doi: 10.11958/20161525

• 实验研究 • 上一篇    下一篇

兔口腔黏膜上皮细胞与 PCL 电纺纤维的复合尿道支架的 制备及可行性分析

关勇，孟庆娅，徐国栋，张富义，舒剑波   

  1. 天津市儿童医院泌尿外科（邮编 300134）
  • 收稿日期:2016-12-13 修回日期:2017-05-01 出版日期:2017-06-15 发布日期:2017-07-05
  • 作者简介:关勇（1971），男，医学博士，主任医师，主要从事儿童泌尿外科疾病治疗研究
  • 基金资助:
    天津市卫生局科技基金重点项目（2013KR08）

The preparation and feasibility study of composite urethral stent-rabbit oral mucosal epithelial cells and PCL electrospun fiber

GUAN Yong, MENG Qing-ya, XU Guo-dong, ZHANG Fu-yi, SHU Jian-bo   

  1. Department of Urology, Tianjin Children’s Hospital, Tianjin 300134, China
  • Received:2016-12-13 Revised:2017-05-01 Published:2017-06-15 Online:2017-07-05

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摘要： 目的 探讨兔口腔黏膜上皮细胞与聚己内酯（PCL）电纺纤维的复合支架用于尿道修复的可行性。方法 用三氯甲烷和无水甲醇体积比 5∶1 的混合液配制 25%PCL 混合液，通过静电纺丝得到 PCL 纤维管状支架，在其管腔 内种植 1.5×105个兔口腔黏膜上皮细胞，然后埋入裸鼠皮下，2 周后取出，通过 HE 和细胞角蛋白抗体免疫荧光染色 对该复合尿道支架进行生物学评价。未种植兔口腔黏膜上皮细胞的 PCL 管为对照组，其评价方法同上。结果 与 对照组比较，种植兔口腔黏膜上皮细胞的 PCL 管展现出良好的细胞化，并且兔口腔黏膜上皮细胞在 PCL 管腔表面形 成了一层致密的细胞层，兔口腔黏膜上皮细胞能够在 PCL 管腔表面存活增殖。结论 兔口腔黏膜上皮细胞可以作 为组织工程化尿道支架的种子细胞来源之一，其与 PCL 电纺纤维相结合，可以构建适合于尿道修复需要的组织工程 替代材料。

关键词: 口腔黏膜, 上皮细胞, 组织工程, 聚己内酯, 尿道支架, 静电纺丝

Abstract: Objective To explore the feasibility of using composite scaffolds of rabbit oral epithelial cells and polycaprolactone (PCL) electrospun fibers for urethral repair. Methods The 25% PCL was prepared using a 5∶1 by volume mixture of trichloromethane and anhydrous methanol, and PCL fiber tubular scaffolds were obtained by electrospinning. Rabbit oral mucosa epithelial cells (1.5×105) were implanted on the PCL scaffold. Subsequently, they were embedded in nude mice subcutaneous, explanted in 2 weeks. PCL fiber tubular scaffolds without rabbit oral mucosa epithelial cells were used as control. The complex urethral scaffolds were evaluated by immunofluorescence staining with cytokeratin antibody and HE staining. Results Compared with blank PCL group, the rabbit oral mucosa epithelial cell group showed a good cellularization. Rabbit oral mucosa epithelial cells formed a dense cell layer on the surface of PCL lumen, which suggested that rabbit oral mucosa epithelial cells can proliferate on the surface of PCL lumen. Conclusion Rabbit oral epithelial cells can be used as one of the seed cells for tissue engineered urethral scaffolds, and it is possible to construct tissue engineering substitute materials for urethral repair by rabbit oral epithelial cells combined with PCL.

Key words: mouth mucosa, epithelial cells, tissue engineering, polycaprolactone, urethral stent, electrospinning