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静电纺丝聚丙烯腈/聚氧化乙烯纳米纤维支架上培养时,间充质干细胞向膀胱平滑肌细胞分化的能力增强。

Improved bladder smooth muscle cell differentiation of the mesenchymal stem cells when grown on electrospun polyacrylonitrile/polyethylene oxide nanofibrous scaffold.

机构信息

Department of Biotechnology, University of Tehran, Tehran, Iran.

Department of Medical Biotechnology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran.

出版信息

J Cell Biochem. 2019 Sep;120(9):15814-15822. doi: 10.1002/jcb.28852. Epub 2019 May 8.

Abstract

Reconstruction of the bladder wall plays an important role in improving its function in patients with urinary bladder dysfunction. Tissue engineering has been trying to introduce biocompatible nanofibers as scaffolds for bladder wall matrix substitutes. In this study a composite nanofibrous scaffold was fabricated from polyacrylonitrile (PAN) and polyethylene oxide (PEO) blend by electrospinning method and then its morphological and mechanical characteristics was evaluated by scanning electron microscopy (SEM), tensile, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Then smooth muscle cell (SMC) differentiation supportive capacity of PAN-PEO nanofibers was investigated by culturing of human adipose tissue-derived mesenchymal stem cells (AT-MSCs) on this scaffold and then its differentiation potential in different groups was investigated using SMC-related gene and protein markers. SEM and MTT results demonstrated that PAN-PEO supported AT-MSCs attachment, growth and proliferation, especially at early times after cell seeding. The obtained results from real-time reverse transcription polymerase chain reaction revealed that collagen-I-α1, collagen-III-α1, α-smooth muscle actin (α-SMA), calponin1, SM22α, caldesmon1, elastin, and myosin heavy chain (MHC) genes were expressed in AT-MSCs cultured on PAN-PEO significantly higher than those stem cells that cultured on the culture plate as a control. In addition α-SMA and MHC proteins were also expressed in AT-MSCs cultured on PAN-PEO significantly higher than control. According to the results PAN-PEO nanofibrous scaffold showed a positive AT-MSCs-seeded PAN-PEO has a great promising potential to use in bladder tissue engineering applications.

摘要

膀胱壁重建对于改善膀胱功能障碍患者的膀胱功能具有重要作用。组织工程一直在尝试引入生物相容性的纳米纤维作为膀胱壁基质替代物的支架。在这项研究中,通过静电纺丝方法从聚丙烯腈(PAN)和聚氧化乙烯(PEO)共混物制备了复合纳米纤维支架,并通过扫描电子显微镜(SEM)、拉伸和 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)试验评估了其形态和机械特性。然后,通过在该支架上培养人脂肪组织来源的间充质干细胞(AT-MSCs),研究了 PAN-PEO 纳米纤维支持平滑肌细胞(SMC)分化的能力,然后使用 SMC 相关基因和蛋白标志物研究了其在不同组中的分化潜力。SEM 和 MTT 结果表明,PAN-PEO 支持 AT-MSCs 的附着、生长和增殖,尤其是在细胞接种后的早期。实时逆转录聚合酶链反应的结果表明,在 PAN-PEO 上培养的 AT-MSCs 中,I 型胶原蛋白-α1、III 型胶原蛋白-α1、α-平滑肌肌动蛋白(α-SMA)、钙调蛋白 1、SM22α、钙调蛋白 1、弹性蛋白和肌球蛋白重链(MHC)基因的表达明显高于作为对照的在培养板上培养的干细胞。此外,在 PAN-PEO 上培养的 AT-MSCs 中也明显表达了 α-SMA 和 MHC 蛋白。根据结果,PAN-PEO 纳米纤维支架显示出阳性的 AT-MSCs 接种结果,因此在膀胱组织工程应用中具有很大的应用潜力。

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