Electrospinning is a popular method for creating random, non-woven fibrous templates for biomedical applications, and a subtype technique termed near-field electrospinning (NFES) was devised by reducing the air gap distance to millimeters. This decreased working distance paired with precise translational motion between the fiber source and collector allows for the direct writing of fibers. We demonstrate a near-field electrospinning device designed from a MakerFarm Prusa i3v three-dimensional (3D) printer to write polydioxanone (PDO) microfibers. PDO fiber diameters were characterized over the processing parameters: Air gap, polymer concentration, translational velocity, needle gauge, and applied voltage. Fiber crystallinity and individual fiber uniformity were evaluated for the polymer concentration and translational fiber deposition velocity. Fiber stacking was evaluated for the creation of 3D templates to guide the alignment of human gingival fibroblasts. The fiber diameters correlated positively with polymer concentration, applied voltage, and needle gauge; and inversely correlated with translational velocity and air gap distance. Individual fiber diameter variability decreases, and crystallinity increases with increasing translational fiber deposition velocity. These data resulted in the creation of tailored PDO 3D templates, which guided the alignment of primary human fibroblast cells. Together, these results suggest that NFES of PDO can be scaled to create precise geometries with tailored fiber diameters for biomedical applications.Polymers 2020, 12, 1 2 of 17 micro-fabricated silicone tip translating at an air gap distance of 5-15 mm over a counter electrode to write an individual fiber [10]. As with TES, this technique utilizes a charged polymer, in solution or melted with heat, to extrude a microfiber on a counter electrode, but the high degree of fiber control with this method arises from positioning the Taylor cone significantly closer to the counter electrode before any bending instabilities can occur. When paired with precise translational movement between the polymer source and collector, fibers can be directly "written" allowing for the creation of specific geometries that can be laid down layer-by-layer. As both NFES and TES utilize a Taylor cone formed in an electric field, it suggests that if a polymer solution/melt has the viscosity, conductivity, and surface tension of TES, then NFES is also possible [11]. To date, numerous polymers have successfully NFES, including but not limited to, polyethylene oxide (PEO), polyvinylpyrrolidone (PVP), polycaprolactone (PCL), polystyrene (PS), and polyvinylidene fluoride (PVDF) [12]. Early NFES setups used an "ink and quill" approach to write fibers onto a grounded substrate. However, NFES have progressed in using motion platforms or modified commercially available three-dimensional (3D) printers to position a spinneret with a continuous polymer source, therefore, allowing continuous fiber deposition with precise placement [10,11,13].In this paper, we inve...