中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (21): 3855-3858.doi: 10.3969/j.issn.1673-8225.2012.21.013

• 复合支架材料 composite scaffold materials • 上一篇    下一篇

胶原海绵支架体外构建组织工程膀胱黏膜***★◆

张  明1，徐明曦1，吴稼晟2，李  伟2，孙  康2，王  忠1，周  哲1，卢慕峻1   

  1. 1上海交通大学医学院附属第九人民医院泌尿外科，上海市  200011；2上海交通大学材料科学与工程学院金属基复合材料国家重点实验室，上海市  200240
  • 收稿日期:2011-10-19 修回日期:2011-12-20 出版日期:2012-05-20 发布日期:2012-05-20
  • 通讯作者: 卢慕峻，副主任医师，硕士生导师，上海交通大学医学院附属第九人民医院泌尿外科，上海市 200011 lumujun@ 163.com 并列通讯作者：孙康，教授，博士生导师，上海交通大学材料科学与工程学院金属基复合材料国家重点实验室，上海市200240 ksun@ sjtu.edu.cn
  • 作者简介:张明★，男，1985年生，山东省胶州市人，汉族，上海交通大学医学院在读硕士，主要从事组织工程泌尿系统修复研究。 zhangm4911@ 163.com 并列第一作者：徐明曦★，男，1976年生，上海市人，汉族，上海交通大学医学院在读硕士，主治医师，主要从事泌尿外科男科工作。xmx197630@ 163.com
  • 基金资助:

    国家自然科学基金面上项目(81070605)；上海交通大学“医工(理)交叉研究基金”(YG2011MS14)；上海市教委科研创新项目(09YZ76)。

Construction of tissue engineering bladder mucosa by collagen sponge in vitro

Zhang Ming1, Xu Ming-xi1, Wu Jia-sheng2, Li Wei2, Sun Kang2, Wang Zhong1, Zhou Zhe1, Lu Mu-jun1   

  1. 1Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai  200011, China; 2State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai  200240, China
  • Received:2011-10-19 Revised:2011-12-20 Online:2012-05-20 Published:2012-05-20
  • Contact: Lu Mu-jun, Associate chief physician, Master’s supervisor, Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China lumujun@163.com Sun Kang, Professor, Doctoral supervisor, State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China ksun@sjtu.edu.cn
  • About author:Zhang Ming★, Studying for master’s degree, Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China zhangm4911@ 163.com Xu Ming-xi★, Studying for master’s degree, Attending physician, Department of Urology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China xmx197630@ 163.com
  • Supported by:

    the National Natural Science Foundation of China, No.81070605*; Biomedical Engineering Research Fund of Shanghai Jiao Tong University, No. YG2011MS14*; Research and Innovation Projects of Shanghai Municipal Education Commission, No.09YZ76*

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摘要：

背景：组织工程膀胱黏膜层的构建在组织工程膀胱修复中占有重要的地位，但目前并没有最合理的构建方法。
目的：探讨胶原海绵支架复合猪膀胱尿路上皮细胞体外构建膀胱黏膜结构的可行性。
方法：刮取猪膀胱黏膜层后用酶消化方式进行猪膀胱尿路上皮细胞原代培养，并进行尿路上皮细胞标志物免疫荧光和RT-PCR鉴定。制备疏松多孔的胶原海绵支架材料，将第3代尿路上皮细胞接种在胶原海绵支架上，体外培养4~8 d后观察尿路上皮细胞和胶原海绵材料的复合情况。
结果与结论：原代培养的猪膀胱尿路上皮细胞呈“多角形”、“铺路石”样，以克隆团形式生长。免疫荧光鉴定AE1/AE3阳性，RT-RCR检测uroplakin-ⅠA、uroplakin-Ⅱ阳性。胶原海绵复合尿路上皮细胞体外培养4~8 d后，细胞在胶原海绵支架上生长良好，覆盖胶原材料的表面并长入材料内部，保持了尿路上皮细胞的特性。体外培养6 d时，尿路上皮细胞与胶原支架复合效果最好，同时细胞数量也最多。结果初步表明了胶原海绵复合膀胱尿路上皮细胞可以构建组织工程膀胱黏膜，且体外培养6 d为最佳时间点。

关键词: 膀胱黏膜, 胶原, 尿路上皮细胞, 组织工程, 支架材料

Abstract:

BACKGROUND: Construction of tissue engineering bladder mucosa is very important in the repair of tissue engineering bladder. But there is no reasonable repair method up to now.
OBJECTIVE: To construct the bladder mucosa by collagen sponge scaffold combined with porcine bladder cells.
METHODS: The urothelial cells were scraped from the porcine urinary bladder mucosa, and digested by trypsin for primary culture. The marker of urothelial cells at passage three were detected by immunofluorescence and reverse transcription-PCR. Porous collagen sponge scaffolds were made. The urothelial cells at passage three were seeded on the collagen sponge scaffolds. After cultured for 4-8 days, the cell-collagen sponge composites were observed.
RESULTS AND CONCLUSION: Primary cultured porcine bladder urothelial cells were polygon and paving stone-like, and they could form the clone groups. Immunofluorescence identification of AE1/AE3 was positive, reverse transcription-RCR detection of uroplakin-IA and uroplakin-Ⅱ were positive. The histological study found that after cultured for 4-8 days, the cells grew well on collagen sponge scaffold, the cells covering the surface of collagen materials and growing into the scaffold maintained the characteristics of urothelial cells. After cultured for 6 days in vitro, the cell-seeded collagen scaffold grew best with the highest number of cells compared to 4 and 8 days. Urothelial cell-seeded collagen sponge is a suitable material for the repair of bladder mucosa by tissue engineering technology in vitro. The best time of culture before replantation in vitro is 6 days.

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