Bladder Acellular Matrix Grafts Seeded with Adipose-Derived Stem Cells and Incubated Intraperitoneally Promote the Regeneration of Bladder Smooth Muscle and Nerve in a Rat Model of Bladder Augmentation
Bladder Acellular Matrix Grafts Seeded with Adipose-Derived Stem Cells and Incubated Intraperitoneally Promote the Regeneration of Bladder Smooth Muscle and Nerve in a Rat Model of Bladder Augmentation
复制标题
膀胱脱细胞基质移植物接种脂肪干细胞并腹膜内孵育促进膀胱增大大鼠模型中膀胱平滑肌和神经的再生
DOI：
10.1089/scd.2015.0246
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发表时间：
2016-03-01
影响因子：
4
通讯作者：
Lu Mujun
中科院分区：
文献类型：
--
作者：
Zhou Zhe;Da Jun;Lu Mujun
本研究旨在探讨膀胱无细胞基质移植物（BAMG）与脂肪干细胞（ASCs）联合腹腔内培养重建膀胱的可行性，并探讨其机制。将接种ASC的（实验组）和未接种ASC的（对照组）BAMG在雄性大鼠腹膜中孵育2周，然后收获用于膀胱增大。对孵育的BAMG的组织学分析显示，两组中均质胶原束中生长有大量细胞。在对照BAMG中，这些细胞是间充质来源的，而在ASC接种的BAMG中，分别在支架的内部和表面上发现肌成纤维细胞和间皮细胞。免疫荧光分析表明，一些肌成纤维细胞转分化的ASCs后，腹腔内孵育2周。术后4周和14周实验组膀胱容量均大于对照组。术后4周和14周实验组和对照组的尿道再生良好，无显著性差异。免疫组化和免疫荧光染色定量结果显示，术后4周和14周实验组平滑肌细胞（SMC）再生情况均明显好于对照组。术后14周实验组神经细胞数量明显增多。术后4周，免疫荧光双标显示BAMG中部分SMC由植入的ASCs转分化而来，但术后14周未检测到ASCs的CM-DiI标记。两者合计，我们的结果表明，ASC接种和腹膜孵育的BAMG不仅促进膀胱平滑肌和神经的形态再生，而且促进膀胱容量，这表明它们具有膀胱再生的潜力。
The objective of this study was to investigate the feasibility of bladder acellular matrix grafts (BAMGs) seeded with adipose-derived stem cells (ASCs) followed by intraperitoneal incubation for bladder reconstruction in a rat model of bladder augmentation, and to explore the underlying mechanism. Autologous CM-DiI-labeled ASC-seeded (experimental group) and unseeded (control group) BAMGs were incubated in the peritoneum of male rats for 2 weeks and then harvested for bladder augmentation. Histological analysis of the incubated BAMGs revealed numerous cells growing in homogeneous collagen bundles in both groups. In the control BAMGs, these cells were mesenchyme derived, while in the ASC-seeded BAMGs, myofibroblasts and mesothelial cells were found inside and on the surface of the scaffold, respectively. Immunofluorescence analysis demonstrated that some of the myofibroblasts were transdifferentiated from the ASCs after 2 weeks of intraperitoneal incubation. The greater bladder capacity was found in the experimental group than the control group both 4 and 14 weeks postoperatively. Histological analysis revealed that the entire urothelium regenerated well both in the experimental group and the control group without significant difference 4 weeks and 14 weeks postoperatively. From the quantitative data of immunohistochemical and immunofluorescence staining, the smooth muscle cells (SMCs) regenerated significantly better in the experimental group than the control group both 4 weeks and 14 weeks postoperatively. Also significantly more nerve cells were found in the experimental group 14 weeks postoperatively. At 4 weeks postoperatively, the immunofluorescence double staining revealed that some SMCs in the BAMG were transdifferentiated from the implanted ASCs, but no CM-DiI labeling of ASCs was detected 14 weeks postoperatively. Taken together, our results demonstrate that ASC-seeded and peritoneally incubated BAMGs promote not only the morphological regeneration of the bladder smooth muscle and nerve, but also the bladder capacity, which indicates their potential for bladder regeneration.