Fabrication of Tissue-Engineered Bionic Urethra Using Cell Sheet Technology and Labeling By Ultrasmall Superparamagnetic Iron Oxide for Full-Thickness Urethral Reconstruction.
Fabrication of Tissue-Engineered Bionic Urethra Using Cell Sheet Technology and Labeling By Ultrasmall Superparamagnetic Iron Oxide for Full-Thickness Urethral Reconstruction.
复制标题
利用细胞片技术和超小型超顺磁性氧化铁标记制造组织工程仿生尿道，用于全层尿道重建
作者：
Zhou S;Yang R;Zou Q;Zhang K;Yin T;Zhao W;Shapter JG;Gao G;Fu Q
尿道狭窄仍然是一个重建的挑战，由于不满意的结果和狭窄复发率高。一个理想的尿道重建应该建立与原来尿道壁相似的结构和功能。我们利用细胞片层技术构建了一种新型的组织工程化仿生骨，并报告了其在犬模型中的存活能力。收集少量口腔和脂肪组织，分离脂肪源性干细胞、口腔粘膜上皮细胞和口腔粘膜成纤维细胞并用于制备细胞片。将细胞片层分层管状化以形成3层组织工程支架，并用超小超顺磁性氧化铁（USPIO）标记。将构建的组织工程化猪尿道皮下移植3周，以促进血管再生和生物力学强度。然后，用组织工程化仿生尿道替代2cm长的管状阴茎尿道。尿道置换术后3个月，MRI可有效检测到USPIO标记的组织工程仿生尿道移植部位。组织学上，回收的仿生肌仍然显示3层，包括上皮层、纤维层和成肌细胞层。皮下移植3周后，免疫荧光分析显示，仿生尿道血管密度在构建初期明显增加，在尿道置换术后3个月进一步上调，接近尿道组织正常水平。我们的研究是第一个实验证明，3层组织工程尿道可以建立使用细胞片技术，并可以促进再生的结构和功能的尿道类似于正常尿道。
Urethral strictures remain a reconstructive challenge, due to less than satisfactory outcomes and high incidence of stricture recurrence. An “ideal” urethral reconstruction should establish similar architecture and function as the original urethral wall. We fabricated a novel tissue-engineered bionic urethras using cell sheet technology and report their viability in a canine model. Small amounts of oral and adipose tissues were harvested, and adipose-derived stem cells, oral mucosal epithelial cells, and oral mucosal fibroblasts were isolated and used to prepare cell sheets. The cell sheets were hierarchically tubularized to form 3-layer tissue-engineered urethras and labeled by ultrasmall super-paramagnetic iron oxide (USPIO). The constructed tissue-engineered urethras were transplanted subcutaneously for 3 weeks to promote the revascularization and biomechanical strength of the implant. Then, 2 cm length of the tubularized penile urethra was replaced by tissue-engineered bionic urethra. At 3 months of urethral replacement, USPIO-labeled tissue-engineered bionic urethra can be effectively detected by MRI at the transplant site. Histologically, the retrieved bionic urethras still displayed 3 layers, including an epithelial layer, a fibrous layer, and a myoblast layer. Three weeks after subcutaneous transplantation, immunofluorescence analysis showed the density of blood vessels in bionic urethra was significantly increased following the initial establishment of the constructs and was further up-regulated at 3 months after urethral replacement and was close to normal level in urethral tissue. Our study is the first to experimentally demonstrate 3-layer tissue-engineered urethras can be established using cell sheet technology and can promote the regeneration of structural and functional urethras similar to normal urethra.
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