Lower back pain (LBP) and neck problems are the most common orthopedic diseases worldwide, and the main reason behind LBP is intervertebral disc degeneration (IVDD). Therefore, specialized therapeutic applications to induce intervertebral disc (IVD) regeneration is a necessity. Here, we report the use of a collagen-mimetic bioactive peptide nanofiber scaffold (Col-PA/E-PA) for the improvement of disc regeneration by recapitulating the structure and function of the natural extracellular matrix (ECM) of intervertebral connective tissue. Following two weeks of degeneration, the bioactive nanofiber scaffold was topically applied and the IVD regeneration process was observed through histochemical analyses. The collagen-mimetic bioactive peptide nanofiber system was found to significantly promote glycosaminoglycan and collagen deposition at the site of injury compared to control nanofiber system and saline groups. In addition, the bioactive scaffold was consistently associated with lower values in degeneration scoring analyses, confirming the functional recovery of the tissue. Overall, the collagen-mimetic peptide nanofiber scaffold was able to prevent the progression of IVD degeneration and provide further functional recovery to the tissue.