Decellularization and sterilization of porcine urinary bladder matrix for tissue engineering in the lower urinary tract
Decellularization and sterilization of porcine urinary bladder matrix for tissue engineering in the lower urinary tract
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DOI：
10.2217/17460751.3.2.145
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发表时间：
2008-03-01
影响因子：
2.7
通讯作者：
MacNeil, Sheila
中科院分区：
文献类型：
--
作者：
Rosario, Derek J.;Reilly, Gwendolen C.;MacNeil, Sheila
背景资料：已经描述了几种合成和天然基质用于膀胱的组织工程，但是关于处理对其随后的机械性能或与人细胞的相互作用的影响的信息很少。目的：我们的目的是评估脱层，脱细胞和灭菌对猪膀胱基质（UBM）的机械性能的影响，然后评估UBM作为人膀胱细胞重建支架的能力。研究方法：在机械分层前后共评估了20个猪膀胱，并通过比较几种脱细胞和终端灭菌方法检查组织学和机械特性，在每个阶段跟踪了4个猪膀胱。将无菌UBM与正常人尿路上皮细胞和膀胱基质细胞同时共培养，或与基质细胞随后与尿路上皮细胞共培养。结果如下：在低渗缓冲液、0.1%十二烷基硫酸钠溶液和0.1%过氧乙酸中对所得膀胱基质进行机械分层、物理冲洗，得到具有可接受的机械性能的UBM，其能够支持尿路上皮和膀胱基质细胞。用环氧乙烷进行的最终灭菌导致基质的相当大的硬化，同时与基质细胞共培养和分层接种支架，然后是上皮细胞，得到了类似的结果，具有良好的初始尿路上皮附着（随后细胞丢失）和缓慢的基质细胞渗透。结论：我们描述了一种脱细胞灭菌的猪UBM，其具有可接受的机械性能，显示出作为用于生产下尿路重建的体外组织工程膀胱补片材料的支架的前景。未来的工作需要关注实现安全上皮附着的条件。
Background: Several synthetic and natural matrices have been described for tissue engineering of bladder but there is little information on the effects of processing on their subsequent mechanical performance or interaction with human cells. Aim: Our aim was to assess the effects of delamination, decellularization and sterilization on the mechanical properties of porcine urinary bladder matrix (UBM) and to then assess the ability of the UBM to act as a scaffold for reconstruction with human bladder cells. Methods: A total of 20 porcine bladders were assessed before and after mechanical delamination and four porcine bladders were followed at every stage through a comparison of several decellularization and terminal sterilization methodologies examining histological and mechanical characteristics. The sterile UBM was seeded with normal human urothelial and bladder stromal cells either as a simultaneous coculture, or with stromal cells followed by urothelial cells. Results: Mechanical delamination, physical rinsing of the resulting bladder stroma in hypotonic buffer, 0.1% sodium dodecyl sulfate solution and 0.1% peracetic acid resulted in an UBM with acceptable mechanical properties capable of supporting urothelial and bladder stromal cells. Terminal sterilization with ethylene oxide resulted in considerable stiffening of the matrix simultaneous coculture and layered seeding of scaffolds with stromal cells followed by epithelial cells gave similar results with good initial urothelial attachment (followed by loss of cells later) and slow stromal cell penetration. Conclusion: We describe a decellularized sterilized porcine UBM with acceptable mechanical properties that shows promise as a scaffold for producing an in vitro tissue-engineered bladder patch material for lower urinary tract reconstruction. Future work now needs to focus on the conditions for achieving secure epithelial attachment.