Tissue regeneration often requires recruitment of different cell types and rebuilding of two or more tissue layers to restore function. Here, we describe the creation of a novel multilayered scaffold with distinct fiber organizations—aligned to unaligned and dense to porous—to template common architectures found in adjacent tissue layers. Electrospun scaffolds were fabricated using a biodegradable, tyrosine‐derived terpolymer, yielding densely‐packed, aligned fibers that transition into randomly‐oriented fibers of increasing diameter and porosity. We demonstrate that differently‐oriented scaffold fibers direct cell and extracellular matrix (ECM) organization, and that scaffold fibers and ECM protein networks are maintained after decellularization. Smooth muscle and connective tissue layers are frequently adjacent in vivo; we show that within a single scaffold, the architecture supports alignment of contractile smooth muscle cells and deposition by fibroblasts of a meshwork of ECM fibrils. We rolled a flat scaffold into a tubular construct and, after culture, showed cell viability, orientation, and tissue‐specific protein expression in the tube were similar to the flat‐sheet scaffold. This scaffold design not only has translational potential for reparation of flat and tubular tissue layers but can also be customized for alternative applications by introducing two or more cell types in different combinations.

中文翻译：

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用于平滑肌和结缔组织层再生的多层支架

组织再生通常需要募集不同类型的细胞并重建两个或多个组织层以恢复功能。在这里，我们描述了一种新型多层支架的创建，该支架具有不同的纤维组织——对齐到未对齐和致密到多孔——以模板化相邻组织层中发现的常见结构。静电纺丝支架是使用可生物降解的酪氨酸衍生三元共聚物制造的，产生密集排列的纤维，这些纤维转变为直径和孔隙率增加的随机定向纤维。我们证明了不同方向的支架纤维指导细胞和细胞外基质 (ECM) 组织，并且支架纤维和 ECM 蛋白网络在脱细胞后得以维持。平滑肌层和结缔组织层在体内经常相邻；我们表明，在单个支架内，该架构支持收缩平滑肌细胞的排列和 ECM 原纤维网的成纤维细胞的沉积。我们将扁平支架卷成管状结构，培养后，管中的细胞活力、方向和组织特异性蛋白质表达与平板支架相似。这种支架设计不仅具有修复扁平和管状组织层的平移潜力，而且还可以通过以不同组合引入两种或多种细胞类型，针对替代应用进行定制。管中的组织特异性蛋白表达与平板支架相似。这种支架设计不仅具有修复扁平和管状组织层的平移潜力，而且还可以通过以不同组合引入两种或多种细胞类型，针对替代应用进行定制。管中的组织特异性蛋白表达与平板支架相似。这种支架设计不仅具有修复扁平和管状组织层的平移潜力，而且还可以通过以不同组合引入两种或多种细胞类型，针对替代应用进行定制。