Direct Writing Electrospinning of Scaffolds with Multidimensional Fiber Architecture for Hierarchical Tissue Engineering

[Image: see text] Nanofibrous structures have long been used as scaffolds for tissue engineering (TE) applications, due to their favorable characteristics, such as high porosity, flexibility, high cell attachment and enhanced proliferation, and overall resemblance to native extracellular matrix (ECM). Such scaffolds can be easily produced at a low cost via electrospinning (ESP), but generally cannot be fabricated with a regular and/or complex geometry, characterized by macropores and uniform thickness. We present here a novel technique for direct writing (DW) with solution ESP to produce complex three-dimensional (3D) multiscale and ultrathin (∼1 μm) fibrous scaffolds with desirable patterns and geometries. This technique was simply achieved via manipulating technological conditions, such as spinning solution, ambient conditions, and processing parameters. Three different regimes in fiber morphologies were observed, including bundle with dispersed fibers, bundle with a core of aligned fibers, and single fibers. The transition between these regimes depended on tip to collector distance (Wd) and applied voltage (V), which could be simplified as the ratio V/Wd. Using this technique, a scaffold mimicking the zonal organization of articular cartilage was further fabricated as a proof of concept, demonstrating the ability to better mimic native tissue organization. The DW scaffolds directed tissue organization and fibril matrix orientation in a zone-dependent way. Comparative expression of chondrogenic markers revealed a substantial upregulation of Sox9 and aggrecan (ACAN) on these structures compared to conventional electrospun meshes. Our novel method provides a simple way to produce customized 3D ultrathin fibrous scaffolds, with great potential for TE applications, in particular those for which anisotropy is of importance.