Abstract Background: Macrophages (Mφ) play a significant role in peripheral nerve regeneration. The objective of this study was to investigate the mechanism of action of macrophage-derived exosomes (Exo) in peripheral nerve injury (PNI). Methods The extraction of exosomes (Exo) following the polarization of macrophages (M?) to either the M1 or M2 phenotype. This was done with the intention of intervening in Schwann cells (SCs), and the ensuing experiments included those utilizing the Cell Counting Kit-8 (CCK8), quantitative real-time polymerase chain reaction (qRT-PCR), flow cytometry, sequencing, and Western blot (WB) techniques. In vivo, the sciatic nerve extrusion model was established and divided into PBS, M1-exo and M2-exo groups, which were injected with phosphate-buffered saline (PBS), M1-exo and M2-exo, respectively, while another normal group was set up. The sciatic nerve function index (SFI), wet weight ratio, and musculo-neural tissue staining were evaluated on a weekly basis in each experimental group following surgical intervention. Results M1-exo facilitated SC migration and upregulated glial-derived nerve growth factor (GDNF), whereas M2-exo promoted SC proliferation and migration and upregulated brain-derived nerve growth factor (BDNF) and nerve growth factor receptor (NGFR). Additionally, the following biomarkers were identified: growth factor (NGF), myelin protein zero (MPZ), glial fibrillary acidic protein (GFAP), nerve growth factor receptor (NGFR), and S100 calcium-binding protein B (S100), among others. In vivo, the SFI, wet weight rate, Masson staining of muscle and nerve tissues, and fluorescence area of nerve axons (marker NF200) and SCs (marker S100) in the M2-exo group at week 4 exhibited superior outcomes compared to those in the PBS group, with all differences being statistically significant. Conversely, no significant differences were observed between the M1-exo group and the PBS group. Additionally, both in vivo and ex vivo experiments demonstrated that M2-exo could facilitate the proliferation of SCs through the phosphorylation of serine/threonine kinase (AKT), which subsequently resulted in the downregulation of cell cycle protein WEE1. Conclusion M2-exo enhances the release of nerve regeneration-related genes from SCs, and it can also promote SCs proliferation and migration, thereby facilitating enhanced nerve regeneration through the AKT-WEE1 pathway. Keywords Macrophage (M?); Exosome (Exo); Schwann cells (SCs); Peripheral nerve injury (PNI); WEE1 Chinese Library Classification Number: R651.3