Fabrication of Multilayered Nanofiber Scaffolds with a Highly Aligned Nanofiber Yarn for Anisotropic Tissue Regeneration

[Image: see text] Nanofibrous scaffolds were widely studied to construct scaffold for various fields of tissue engineering due to their ability to mimic a native extracellular matrix (ECM). However, generally, an electrospun nanofiber exhibited a two-dimensional (2D) membrane form with a densely packed structure, which inhibited the formation of a bulk tissue in a three-dimensional (3D) structure. The appearance of a nanofiber yarn (NFY) made it possible to further process the electrospun nanofiber into the desired fabric for specific tissue regeneration. Here, poly(l-lactic acid) (PLLA) NFYs composed of a highly aligned nanofiber were prepared via a dual-nozzle electrospinning setup. Afterward, a noobing technique was applied to fabricate multilayered scaffolds with three orthogonal sets of PLLA NFYs, without interlacing them. Thus the constituent NFYs of the fabric were free of any crimp, apart from the binding yarn, which was used to maintain the integrity of the noobing scaffold. Remarkably, the highly aligned PLLA NFY expressed strengthened mechanical properties than that of a random film, which also promoted the cell adhesion on the NFY scaffold with unidirectional topography and less spreading bodies. In vitro experiments indicated that cells cultured on a noobing NFY scaffold showed a higher proliferation rate during long culture period. The controllable pore structure formed by the vertically arrayed NFY could allow the cell to penetrate through the thickness of the 3D scaffold, distributed uniformly in each layer. The topographic clues guided the orientation of H9C2 cells, forming tissues on different layers in two perpendicular directions. With NFY as the building blocks, noobing and/or 3D weaving methods could be applied in the fabrication of more complex 3D scaffolds applied in anisotropic tissues or organs regeneration.