The development of multilayered anisotropic scaffolds with a 3D anisotropy similar to that of native cardiac tissue with layer-specific oriented multiple cell layers is a major challenge in the reconstruction of engineered cardiac tissue in vitro. Herein, we present an electrospun align-random distinct, heterogeneous nanofiber (NF) mat to facilitate the construction of a multilayered anisotropic scaffold with different orientations, which recreates the cardiac anisotropy via hollow patterned electrolyte-assisted electrospinning (HP-ELES). Based on the HP-ELES process, a distinct but heterogeneous polycaprolactone (PCL) NF mat, which comprised an ultra-thin (<10 μm) aligned NF membrane at the core and a thick random NF mat at the peripheral rim, was readily fabricated. The fabricated heterogeneous NF mat was endowed with bifunctional properties of not only uniaxially aligned topographical cues to align the cardiomyocytes (CMs) followed by creating a uniaxial contractile motion but also mechanical stability to enable manual manipulation provided by the aligned NF membrane and random NF mat, respectively. Intriguingly, the in vitro cell culture of CMs possessing spontaneous contraction on the heterogeneous NF mat demonstrated cell alignment and subsequent uniaxial contraction along with aligned NFs. The stacking triple layers also exhibited multiaxial contraction, which potentially simulates the squeezing force of the heart tissue. HP-ELES is highly expected to increase the use of ultra-thin aligned NF membranes for the development of in vitro anisotropic organ models and for in vivo tissue regeneration requiring multilayered anisotropic scaffolds owing to the ease of the manual manipulation of the membrane.

开发具有类似于天然心脏组织的 3D 各向异性的多层各向异性支架，具有特定层定向的多个细胞层，是体外工程心脏组织重建的主要挑战. 在此，我们提出了一种电纺排列随机不同的异质纳米纤维 (NF) 垫，以促进构建具有不同方向的多层各向异性支架，通过中空图案电解质辅助静电纺丝 (HP-ELES) 重建心脏各向异性。基于 HP-ELES 工艺，很容易制造出一种独特但异质的聚己内酯 (PCL) NF 垫，该垫由超薄（<10 μm）对齐的 NF 膜在核心处和厚的随机 NF 垫在外围边缘组成. 制造的异质 NF 垫具有双功能特性，不仅具有单轴对齐的地形线索以对齐心肌细胞（CM），然后产生单轴收缩运动，而且还具有机械稳定性，以实现由对齐的 NF 膜和随机 NF 垫提供的手动操作，分别。耐人寻味的是，在异质 NF 垫上具有自发收缩的 CM 的体外细胞培养证明了细胞排列和随后的单轴收缩以及排列的 NF。堆叠的三层还表现出多轴收缩，这可能模拟心脏组织的挤压力。由于易于手动操作膜，HP-ELES 有望增加超薄对齐 NF 膜的使用，以开发体外各向异性器官模型和需要多层各向异性支架的体内组织再生。