Fibrous PHBV cross-ply scaffolds were fabricated using the electrospinning technique. The electrospun fibers were arranged depending on angles of alignment, which were 180 degrees, 90 degrees, 60 degrees, and 45 degrees. The stress and strain values of the fibrous PHBV cross-ply scaffolds increased as the cross-ply angle increased. At 180 degrees, the strength and strain values of the fibers depended on tensile loading directions. At an alignment of 90 degrees, the PHBV scaffolds had a stress value of 3.5 MPa, which was more than two times higher than the random structure. The cell morphology and proliferation of L-929 cells was strongly dependant on the fiber alignment and the best results were observed when the angle alignment was high. The results of this study showed that the cross-ply structure of the PHBV scaffold affected not only the cell adhesion and spreading properties but also dictated the mechanical properties, which were dependent on the angles of alignment.