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UC-VEGF-SMC三维(3D)纳米支架在膀胱损伤中表现出良好的修复功能。

UC-VEGF-SMC Three Dimensional (3D) Nano Scaffolds Exhibits Good Repair Function in Bladder Damage.

作者信息

Ling Qing, Wang Tao, Yu Xiao, Wang Shao-Gang, Ye Zhang-Qun, Liu Ji-Hong, Yang Sen-Wu, Zhu Xiong-Bo, Yu Jin

出版信息

J Biomed Nanotechnol. 2017 Mar;13(3):313-23. doi: 10.1166/jbn.2017.2343.

Abstract

We constructed a UC-VEGF-SMC three dimensional (3D) scaffold to explore its effect on blood vessel regeneration and bladder repair function in a rabbit model with bladder injury. Rabbit adipose tissue-derived stem cells (ADSCs) were cultured to construct pluripotent stem cell systems that can be induced to differentiate into urothelial cells (UCs) and smooth muscle cells (SMCs). Reverse transcriptase-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, MTT assay and HE staining were used in our study. Rabbit models were divided into an experimental group, control group and sham group. The bladder histology, urodynamics, smooth muscle function, stent degradation rate, urothelial permeability and biomechanical determination of rabbits were detected after grafting the scaffold. Correct stem cells based on the ADSC surface marker and found that CD90 and CD105 were positive and that CD34 and CD45 were negative. RT-PCR showed that ADSC-iPS cells expressed the marker gene of embryonic stem cells (ESCs), which indicated that Sox2, Klf4, Oct4 and c-Myc were inserted into the iPS nucleus and that the ADSC-iPS system was constructed successfully. Immunofluorescence and MTT assays indicated that iPS differentiated into mature SMCs and UCs. ELISA and HEMC culturing methods revealed that vascular endothelia growth factor (VEGF) could promote the growth of HMECs. Rabbit bladder repair function (urodynamics, smooth muscle function, urothelial permeability and biomechanical determination) was stronger in the experimental group than in the control group. UC-VEGF-SMC 3D nano scaffold exhibits good repair function for bladder damage, which may helpful for treatment of damaged bladders.

摘要

我们构建了一种UC-VEGF-SMC三维(3D)支架,以探讨其在兔膀胱损伤模型中对血管再生和膀胱修复功能的影响。培养兔脂肪组织来源的干细胞(ADSCs),构建可诱导分化为尿路上皮细胞(UCs)和平滑肌细胞(SMCs)的多能干细胞系统。本研究采用逆转录聚合酶链反应(RT-PCR)、酶联免疫吸附测定(ELISA)、免疫荧光、MTT法和苏木精-伊红(HE)染色。兔模型分为实验组、对照组和假手术组。植入支架后,检测兔的膀胱组织学、尿动力学、平滑肌功能、支架降解率、尿路上皮通透性和生物力学。基于ADSC表面标志物鉴定出正确的干细胞,发现CD90和CD105呈阳性,CD34和CD45呈阴性。RT-PCR显示ADSC-iPS细胞表达胚胎干细胞(ESCs)的标志物基因,这表明Sox2、Klf4、Oct4和c-Myc已插入iPS细胞核,成功构建了ADSC-iPS系统。免疫荧光和MTT分析表明iPS可分化为成熟的SMC和UC。ELISA和HEMC培养方法显示血管内皮生长因子(VEGF)可促进人微血管内皮细胞(HMECs)的生长。实验组兔的膀胱修复功能(尿动力学、平滑肌功能、尿路上皮通透性和生物力学测定)强于对照组。UC-VEGF-SMC 3D纳米支架对膀胱损伤具有良好的修复功能,可能有助于治疗受损膀胱。

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