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高度取向的微纤维聚酯聚氨酯支架上的骨骼肌生成

Skeletal myogenesis on highly orientated microfibrous polyesterurethane scaffolds.

作者信息

Riboldi S A, Sadr N, Pigini L, Neuenschwander P, Simonet M, Mognol P, Sampaolesi M, Cossu G, Mantero S

机构信息

Department of Bioengineering, Politecnico di Milano, 20133 Milano, Italy.

出版信息

J Biomed Mater Res A. 2008 Mar 15;84(4):1094-101. doi: 10.1002/jbm.a.31534.

Abstract

Skeletal myogenesis is a complex process, which is known to be intimately depending on an optimal outside-in substrate-cell signaling. Current attempts to reproduce skeletal muscle tissue in vitro using traditional scaffolds mainly suffer from poor directionality of the myofibers, resulting in an ineffective vectorial power generation. In this study, we aimed at investigating skeletal myogenesis on novel biodegradable microfibrous scaffolds made of DegraPol, a block polyesterurethane previously demonstrated to be suitable for this application. DegraPol was processed by electrospinning in the form of highly orientated ("O") and nonorientated ("N/O") microfibrous meshes and by solvent-casting in the form of nonporous films ("F"). The effect of the fiber orientation at the scaffold surface was evaluated by investigating C2C12 and L6 proliferation (via SEM analysis and alamarBlue test) and differentiation (via RT-PCR analysis and MHC immunostaining). We demonstrated that highly orientated elastomeric microfibrous DegraPol scaffolds enable skeletal myogenesis in vitro by aiding in (a) myoblast adhesion, (b) myotube alignment, and (c) noncoplanar arrangement of cells, by providing the necessary directional cues along with architectural and mechanical support.

摘要

骨骼肌生成是一个复杂的过程,已知其紧密依赖于最佳的由外而内的底物 - 细胞信号传导。目前使用传统支架在体外重现骨骼肌组织的尝试主要存在肌纤维方向性差的问题,导致产生无效的矢量动力。在本研究中,我们旨在研究在由DegraPol制成的新型可生物降解微纤维支架上的骨骼肌生成,DegraPol是一种嵌段聚酯聚氨酯,先前已证明适用于此应用。DegraPol通过静电纺丝加工成高度取向(“O”)和非取向(“N/O”)的微纤维网,并通过溶剂浇铸加工成无孔膜(“F”)。通过研究C2C12和L6细胞的增殖(通过扫描电子显微镜分析和alamarBlue测试)以及分化(通过逆转录 - 聚合酶链反应分析和肌球蛋白重链免疫染色)来评估支架表面纤维取向的影响。我们证明,高度取向的弹性微纤维DegraPol支架通过协助(a)成肌细胞粘附、(b)肌管排列和(c)细胞的非共面排列,通过提供必要的方向线索以及结构和机械支持,能够在体外实现骨骼肌生成。

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