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利用骨髓间充质干细胞种植于弹性聚(1,8-辛二醇-柠檬酸酯)基薄膜上实现的膀胱平滑肌再生。

Urinary bladder smooth muscle regeneration utilizing bone marrow derived mesenchymal stem cell seeded elastomeric poly(1,8-octanediol-co-citrate) based thin films.

机构信息

Children's Memorial Hospital Chicago, Division of Pediatric Urology, 2300 Children's Plaza, Chicago, IL 60614, USA.

出版信息

Biomaterials. 2010 Aug;31(24):6207-17. doi: 10.1016/j.biomaterials.2010.04.054. Epub 2010 May 21.

DOI:10.1016/j.biomaterials.2010.04.054
PMID:20488535
Abstract

Bladder regeneration studies have yielded inconclusive results possibly due to the use of unfavorable cells and primitive scaffold design. We hypothesized that human mesenchymal stem cells seeded onto poly(1,8-octanediol-co-citrate) elastomeric thin films would provide a suitable milieu for partial bladder regeneration. POCfs were created by reacting citric acid with 1,8-octanediol and seeded on opposing faces with human MSCs and urothelial cells; normal bladder smooth muscle cells and UCs, or unseeded POCfs. Partial cystectomized nude rats were augmented with the aforementioned POCfs, enveloped with omentum and sacrificed at 4 and 10 weeks. Isolated bladders were subjected to Trichrome and anti-human gamma-tubulin, calponin, caldesmon, smooth muscle gamma-actin, and elastin stainings. Mechanical testing of POCfs revealed a Young's modulus of 138 kPa with elongation 137% its initial length without permanent deformation demonstrating its high uniaxial elastic potential. Trichrome and immunofluorescent staining of MSC/UC POCf augmented bladders exhibited typical bladder architecture with muscle bundle formation and the expression and retention of bladder smooth muscle contractile proteins of human derivation. Quantitative morphometry of MSC/UC samples revealed muscle/collagen ratios approximately 1.75x greater than SMC/UC controls at 10 weeks. Data demonstrate MSC seeded POCfs support partial regeneration of bladder tissue in vivo.

摘要

膀胱再生研究的结果尚无定论,这可能是由于使用了不合适的细胞和原始支架设计。我们假设,将人骨髓间充质干细胞接种到聚(1,8-辛二醇-柠檬酸酯)弹性薄膜上,可为部分膀胱再生提供合适的环境。通过柠檬酸与 1,8-辛二醇反应制备 POCfs,将人骨髓间充质干细胞和尿路上皮细胞接种到 POCfs 的相对面上;将正常膀胱平滑肌细胞和 UCs 或未接种 POCfs 的细胞接种到 POCfs 上。将上述 POCfs 包裹在大网膜中,然后对部分去膀胱的裸鼠进行扩增,并在 4 和 10 周时处死。将分离的膀胱进行三色和抗人微管蛋白γ、钙调蛋白、钙调蛋白、平滑肌γ-肌动蛋白和弹性蛋白染色。POCfs 的机械测试显示杨氏模量为 138kPa,伸长率为初始长度的 137%,没有永久变形,证明其具有很高的单轴弹性潜力。MSCs/UC POCf 扩增膀胱的三色和免疫荧光染色显示出典型的膀胱结构,形成肌束,并表达和保留源自人类的膀胱平滑肌收缩蛋白。MSCs/UC 样本的定量形态计量学分析显示,在 10 周时,肌肉/胶原比大约是 SMC/UC 对照的 1.75 倍。数据表明,MSCs 接种的 POCfs 支持体内部分膀胱组织的再生。

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