Aligned fiber scaffolds can mimic the parallel-aligned fibrils in the extracellular matrix (ECM) and regulate many cellular behaviors. Generally, the aligned fibers are developed by a microplate micropatterned method, freeze casting, evaporation, slip casting, ice-templating, plain weaving, and fiber forming techniques. In this study, the Forcespinning® (FS) technology was used to prepare gelatin:zein protein fiber scaffolds with an average diameter of 400 ± 9.6 nm. Based on the collection process, aligned fiber scaffolds were manufactured by FS. Improved mechanical properties were observed. Cell viability, cell adhesion, proliferation, and drug release properties were measured. The cell viability was studied with human fibroblast cells and a low cytotoxic effect was observed on the cells at the highest ratio of gelatin:zein aligned fibers. The cells were aligned on the surface of the linear fiber scaffolds. Berberine chloride release was measured and sustained release rate was observed over the 15 days. Collectively, the morphological features, manipulable architecture, prolonged Berberin release and cytotoxicity of fiber scaffolds suggest that they could be useful in the tissue engineering, skin regeration, hernia repair, and drug delivery.