Silk-based materials have great potentials in medical applications because of the proper mechanical properties and outstanding biocompatibility. This paper reports a novel method to prepare multi-layered silk fibroin (SF) tissue engineering scaffolds with well-aligned fibers oriented in different angles by an electrospinning process. The resulted aligned fibers construct the scaffolds with a grill-like structure, large area and improved mechanical properties, which have been demonstrated by scanning electron microscopy and tensile test. Biocompatibility analysis proves that the grill-like structure well guides the adhesion, proliferation and differentiation of porcine iliac endothelial cells along the aligned SF fibers in different directions.