Spinal cord injury (SCI) creates an adverse environment for axon regeneration. As a result, the axons at the injury sites begin to be atrophy, retract and lose their functions. Several strategies to promote axon regeneration at the injury site have been tested, but the progress is very limited. One of the major reasons is that the regenerated axons often extend randomly and do not reach the proper place. Fabricating linearly ordered materials as nerve guidance would be important to solve such problems. In this study, a novel type of nerve guidance material was prepared from the bovine aponeurosis, which mainly consisted of ordered collagen fibers. The processed material showed good cell compatibility and low immunogenisity. Moreover, the processed material guided the neurites outgrowth of in vitro cultured cortical neurons along its fibers. The results suggested that the processed aponeurosis would be a proper nerve guidance biomaterial for SCI repair.