In this study, nonwoven gelatin (Gel) fabrics crosslinked usingN-acetyl-D-glucosamine (GlcNAc) were characterized and compared with those crosslinked using methylglyoxal (MG) and by thermal dehydration. We prepared Gel with 25% concentration along with Gel/GlcNAc and Gel/MG with a GlcNAc-to-Gel ratio of 5% and MG-to-Gel ratio of 0.6%. A high voltage of 23 kV, solution temperature of 45 °C, and distance of 10 cm between the tip and the collector were applied during electrospinning. The electrospun Gel fabrics were crosslinked by heat treatment at 140 and 150 °C for 1 d. The electrospun Gel/GlcNAc fabrics were treated at 100 and 150 °C for 2 d, while the Gel/MG fabrics were heat-treated for 1 d. The Gel/MG fabrics exhibited higher tensile strength and lower elongation than the Gel/GlcNAc fabrics. Overall, Gel/MG crosslinked at 150 °C for 1 d showed a significant enhancement in tensile strength, high hydrolytic degradation, and excellent biocompatibility, with cell viability percentages of 105 and 130% at 1 and 3 d, respectively. Therefore, MG is a promising Gel crosslinker.