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Solution fibre spinning technique for the fabrication of tuneable decellularised matrix-laden fibres and fibrous micromembranes

机译:溶液纤维纺丝技术,用于制造可调性脱细胞纤维和纤维状微调的纤维

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Recreating tissue-specific microenvironments of the extracellular matrix (ECM) in vitro is of broad interest for the fields of tissue engineering and organ-on-a-chip. Here, we present biofunctional ECM protein fibres and suspended membranes, with tuneable biochemical, mechanical and topographical properties. This soft and entirely biologic membrane scaffold, formed by micro-nano-fibres using low voltage electrospinning, displays three unique characteristics for potential cell culture applications: high-content of key ECM proteins, single-layered mesh membrane, and flexibility for in situ integration into a range of device setups. Extracellular matrix (ECM) powder derived from urinary bladder, was used to fabricate the ECM-laden fibres and membranes. The highest ECM concentration in the dry protein fibre was 50 wt%, with the rest consisting of gelatin. Key ECM proteins, including collagen IV, laminin, and fibronectin, were shown to be preserved post the biofabrication process. The single fibre tensile Young's modulus can be tuned for over two orders of magnitude between similar to 600 kPa and 50 MPa depending on the ECM content. Combining the fibre mesh printing with 3D printed or microfabricated structures, culture devices were constructed for endothelial layer formation, and a trans-membrane co-culture formed by glomerular cell types of podocytes and glomerular endothelial cells, demonstrating feasibility of the membrane culture. Our cell culture observation points to the importance of membrane mechanical property and re-modelling ability as a factor for soft membrane-based cell cultures. The ECM-laden fibres and membranes presented here would see potential applications in in vitro assays, and tailoring structure and biological functions of tissue engineering scaffolds.
机译:在体外重建细胞外基质(ECM)的组织特异性微环境对于组织工程和芯片器官的田地具有广泛的兴趣。在此,我们呈现了生物功能的ECM蛋白纤维和悬浮膜,具有可调的生物化学,机械和地形特性。这种柔软和完全的生物膜支架,通过使用低压静电纺丝形成的微纳米纤维,显示出潜在的细胞培养应用的三种独特的特性:高含量的关键ECM蛋白,单层网膜,以及原位整合的灵活性进入一系列设备设置。衍生自膀胱的细胞外基质(ECM)粉末用于制造ECM-载纤维和膜。干蛋白纤维中最高的ECM浓度为50wt%,其余部分由明胶组成。将键ECM蛋白质,包括胶原蛋白IV,层粘连蛋白和纤连蛋白,被证明可以保存生物结缔组件。根据ECM含量,可以在与600kPa和50MPa类似的情况下调谐单个纤维拉伸杨氏模量超过两个数量级。将具有3D印刷或微制造结构的纤维网印刷结合,构建培养装置,用于内皮层形成,以及通过肾小球细胞类型的肾小球细胞和肾小球内皮细胞形成的跨膜共培养,证明了膜培养的可行性。我们的细胞培养观察点指向膜力学性能的重要性和重新建模能力作为基于软膜的细胞培养的因素。这里呈现的ECM-升起的纤维和膜将在体外测定中看到潜在的应用,以及组织工程支架的剪裁结构和生物学功能。

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