Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA.
BJU Int. 2012 Jul;110(2 Pt 2):E106-12. doi: 10.1111/j.1464-410X.2011.10845.x. Epub 2012 Jan 31.
Re-epithelialization demucosa stomach patch is important to prevent the patch being exposed to urine that might cause patch shrinkage and fibrosis formation due to urine-derived chemical irritation. Additionally, Botox A acts by blocking the transmission of nerve impulses to smooth muscles and so paralysing the muscles, which is commonly used to relax muscle for treatment of oesophageal achalasia due to overactive smooth muscle and sphincters of gastrointestinal tract. We fabricated in vitro tissue engineered urothelial mucosa with multi-layers of urothelium and smooth muscle layers seeded on SIS scaffold and then used this cell-scaffold construct to cover nuke gastro patch combining with Botox A for gastrocystoplasty in a canine model.
To evaluate the demucosalized stomach patch covered with tissue-engineered urothelium for gastrocystoplasty and to determine whether or not injections of Botox A into the re-epithelialized stomach patch can protect the graft from contraction in a canine bladder reconstruction model.
Gastrocystoplasty was performed in 10 adult beagles after hemi-cystectomy using five types of stomach patch (n = 2 per group): entire stomach patches (group I); demucosalized patches (group II); demucosalized patches covered with cell-free small intestinal submucosa (SIS) (group III); demucosalized patches with urothelial and smooth muscle cell-seeded SIS (group IV); and demucosalized patches with the cell-seeded SIS combined with injections of Botox A (group V). The bladder volume/pressure and the graft sizes were measured before surgery and again 10 weeks after bladder augmentation. The graft tissues were examined both histologically and using immunohistochemistry.
All dogs survived and their gastric grafts were all vital with a good blood supply. Gastric metaplasia of urothelium appeared on the top of stomach mucosa patches in two animals in group I. There was calcification formation at the centre of the graft in one animal in group II. As compared with urothelium that was partially covered over with stomach patches in groups II and III, stratified urothelium completely covered the demucosalized gastric patches in groups IV and V. There was less shrinkage of the stomach grafts in groups I and V, which shrank to half of their original size, than of the stomach grafts in groups II, III, and IV, which shrank significantly to one-quarter of their original sizes.
Botox A injections appear to protect the graft contraction in the re-epithelialized stomach flaps. The gastrocystoplasty using demucosalized patches covered with tissue-engineered urothelial mucosa combined with an injection of Botox A could have clinical potential for use in bladder reconstruction.
评估经组织工程化尿路上皮覆盖的去黏膜胃补丁用于胃囊成形术，并确定在犬膀胱重建模型中向再上皮化的胃补丁中注射肉毒毒素 A 是否可以保护移植物免受收缩。
在 10 只成年比格犬中进行了半膀胱切除术后的胃囊成形术，使用了 5 种胃补丁（每组 2 个，n = 2）：完整的胃补丁（I 组）；去黏膜补丁（II 组）；无细胞小肠黏膜下层（SIS）覆盖的去黏膜补丁（III 组）；尿路上皮和平滑肌细胞种植 SIS 的去黏膜补丁（IV 组）；以及种植细胞的 SIS 与肉毒毒素 A 注射联合的去黏膜补丁（V 组）。在手术前和膀胱增大 10 周后再次测量膀胱容积/压力和移植物大小。通过组织学和免疫组织化学检查移植组织。
所有狗均存活，其胃移植物均具有良好的血液供应且具有生命力。I 组中的 2 只动物胃黏膜补丁的顶部出现了尿路上皮化生。II 组中的 1 只动物的移植物中心有钙化形成。与 II 组和 III 组中部分被胃补丁覆盖的尿路上皮相比，IV 组和 V 组中完全去除黏膜的胃补丁上覆盖有分层尿路上皮。与 II 组、III 组和 IV 组相比，I 组和 V 组胃移植物的收缩程度较小，胃移植物收缩至原始大小的一半，而 II 组、III 组和 IV 组的胃移植物明显收缩至原始大小的四分之一。
肉毒毒素 A 注射似乎可以防止再上皮化的胃皮瓣收缩。使用组织工程化尿路上皮覆盖的去黏膜胃补丁进行胃囊成形术，并联合注射肉毒毒素 A，可能具有用于膀胱重建的临床潜力。
