Rosario Derek J, Reilly Gwendolen C, Ali Salah Emadaldeen, Glover Maggie, Bullock Anthony J, Macneil Sheila
Academic Urology Unit, University of Sheffield, Clinical Sciences (South), K Floor, Royal Hallamshire Hospital, Sheffield S10 2JF, UK.
Regen Med. 2008 Mar;3(2):145-56. doi: 10.2217/17460751.3.2.145.
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.
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.
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.
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.
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.
已经描述了几种用于膀胱组织工程的合成和天然基质，但关于加工对其后续机械性能或与人类细胞相互作用的影响的信息很少。
我们的目的是评估分层、脱细胞和灭菌对猪膀胱基质（UBM）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。
对20个猪膀胱在机械分层前后进行评估，并通过比较几种脱细胞和最终灭菌方法，观察四个猪膀胱在每个阶段的组织学和机械特征。将无菌的UBM接种正常人尿道上皮细胞和膀胱基质细胞，要么同时共培养，要么先接种基质细胞，然后接种尿道上皮细胞。
机械分层、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物理冲洗，得到了具有可接受机械性能的UBM，能够支持尿道上皮细胞和膀胱基质细胞。环氧乙烷最终灭菌导致基质显著变硬。支架同时共培养以及先接种基质细胞后接种上皮细胞的分层接种产生了相似的结果，初始尿道上皮附着良好（随后细胞丢失），基质细胞渗透缓慢。
我们描述了一种具有可接受机械性能的脱细胞灭菌猪UBM，它有望作为一种支架，用于生产用于下尿路重建的体外组织工程膀胱贴片材料。现在未来的工作需要集中在实现牢固上皮附着的条件上。
