Wallis M Chad, Yeger Herman, Cartwright Lisa, Shou Zhiping, Radisic Milica, Haig Jennifer, Suoub Mohammed, Antoon Roula, Farhat Walid A
Department of Surgery, Division of Urology, University of Toronto, Hospital for Sick Children, Ontario, Canada.
Tissue Eng Part A. 2008 Mar;14(3):339-48. doi: 10.1089/tea.2006.0398.
We have devised a bioreactor to simulate normal urinary bladder dynamics. The design permits a cell-seeded scaffold made from a modified porcine acellular matrix to be placed between 2 closed chambers filled with culture medium and be mechanically stimulated in a physiologically relevant manner. Specifically designed software increased hydrostatic pressure from 0 to 10 cm of water in a linear fashion in 1 chamber, resulting in mechanical stretch and strain on the scaffold. Pressure was increased over 55 min (filling) and then decreased to 0 over 10 s (voiding). Commercially available small intestinal submucosa scaffolds were used to test the mechanical capabilities of the bioreactor, and pressure waveforms were generated for up to 18 h. Scaffolds were seeded with bladder smooth muscle or urothelial cells and incubated in the bioreactor, which generated pressure waveforms for 6 h. Scaffold integrity was preserved as seen through Masson's trichrome staining. No obvious contamination of the system was noted. Hematoxylin and eosin staining showed presence of cells after incubation in the bioreactor, and immunohistochemistry and real-time reverse transcriptase polymerase chain reaction suggested continued cellular activity. Cellular orientation tended to be perpendicular to the applied pressure. Preliminary results suggest that our bioreactor is a suitable model for simulating normal physiological conditions of bladder cycling in an ex vivo system.
我们设计了一种生物反应器来模拟正常膀胱动力学。该设计允许将由改良的猪脱细胞基质制成的接种细胞的支架放置在两个充满培养基的封闭腔室之间,并以生理相关的方式进行机械刺激。专门设计的软件使一个腔室中的静水压力以线性方式从0增加到10厘米水柱,从而对支架产生机械拉伸和应变。压力在55分钟内增加(充盈),然后在10秒内降至0(排空)。使用市售的小肠黏膜下层支架来测试生物反应器的机械性能,并生成长达18小时的压力波形。将膀胱平滑肌或尿路上皮细胞接种到支架上,并在生物反应器中孵育,生物反应器会产生6小时的压力波形。通过Masson三色染色可见支架完整性得以保留。未发现系统有明显污染。苏木精-伊红染色显示在生物反应器中孵育后有细胞存在,免疫组织化学和实时逆转录聚合酶链反应表明细胞仍有活性。细胞取向倾向于与施加的压力垂直。初步结果表明我们的生物反应器是在体外系统中模拟膀胱循环正常生理条件的合适模型。