Background Conservative approaches and surgical methods have been tried for the treatment of nasal septum perforations (NSPs), but a satisfactory method has not yet been determined. Objective Our study aimed to investigate the effect of platelet-rich fibrin (PRF) and hyaluronic acid (HA) in repairing NSPs, which were experimentally created in rabbit septum. Methods A total of 36 white New Zealand rabbits were included in the study. Perforations measuring 0.7 × 0.7 cm were created in their nasal septa. No additional intervention was made to the control group, which was the first group. For the second, third, and fourth groups, respectively, HA, PRF, and HA + PRF were used in the NSP region. Macroscopic and histopathological evaluations were performed after 40 days. Results In the first group, closure was observed in none of the rabbits’ NSPs. In the second group, 6 rabbits (66.7%) had full closure in their NSPs. In the third group, the NSP of 6 rabbits (66.7%) was completely closed. In the fourth group, the NSP of 7 rabbits (77.8%) had full closure. Conclusion Statistically significant closure was achieved with PRF and/or HA in rabbits in which NSP was established. These materials can be used to increase the likelihood of perforations closing.