Purpose To investigate the effect of platelet-rich fibrin (PRF) on post-operative wound healing following strabismus surgery in a rabbit model. Methods Nine New Zealand white rabbits were involved in the study. One of these nine rabbits was kept in control without having any operation. Both eyes of eight rabbits underwent superior rectus muscle (SRM) resection. After resection of SRM, PRF was applied to SRM of the right eyes. In the left eye, SRM was not wrapped with PRF and served as a control. The rabbits were sacrificed and enucleated 6 weeks after the surgery. Inflammation and vascular proliferation were assessed by staining with haematoxylin-eosin. Scar and fibrosis were examined by the Masson trichrome staining. Immunohistochemical staining was performed for a vascular endothelial growth factor (VEGF) and alpha-smooth muscle actin (alpha-SMA). Results There was no significant difference in terms of inflammation (p = 0.535), vascularization (p = 0.602), and fibrosis (p = 0.745) between the eyes. Immunohistochemical staining for VEGF demonstrated no significant difference (p = 0.745). However, significant staining for alpha-SMA was detected in PRF-treated eyes compared with control eyes (p = 0.037). Conclusion Consistent with the hypothesis that PRF facilitates wound healing as a biocatalyst, there was only a significant difference for alpha-SMA related to myofibroblast activity which is important for wound remodelling. Future comprehensive studies are needed to extend these results.