Aims: This study aimed to evaluate the efficacy of Amniotic Membrane application in rats with Spinal Cord Injury induced by transverse hemisection using kinematic analysis and Sciatic Functional Index. Study design: True experimental research design. Place and Duration of Study: Instiuto de Pesquisa e Desenvolvimento (IP&D) of Universidade do Vale do Paraíba (UNIVAP), between September 2016 and December 2017. Methodology: Fifteen adult male rats were used, allocated into three equal groups: Control (the spinal cord injury and Amniotic Membrane application were simulated), Lesion (spinal cord injury not treated), Amniotic membrane (spinal cord injury treated by amniotic membrane). All animals underwent surgical procedures. A transverse hemisection was performed in groups Lesion and Amniotic Membrene. A fragment of the biomaterial was applied in group AM covering the hemisection area. Results: Sciatic Functional Index and motion analysis were performed by comparing images taken at pre- and postoperative time at 7, 14, 21, and 28 days. The kinematic analysis showed a significant difference between groups Control and Lesion at 7 days (p = 0.023) and 14 days (p = 0.015), and between groups Lesion and Amniotic Membrane at 14 days (p = 0.039), comparing the postoperative periods. The Sciatic Functional Index revealed significant differences between Groups Control and Lesion at 7 (p = -0.002), 14 (p = 0.003), and 21 days (p = 0.009), between Groups Control and Amniotic Membrane at 7 (p = 0.014), 14 (p = 0.007), and 28 days (p = 0; 013), and between Groups Lesion and Amniotic Membrane only at 14 days (p = 0.039). Conclusion: Application of amniotic membrane in spinal cord hemisection in rats induced gait recovery and improvement in SFI compared to the untreated group.