This study evaluated the efficacy of lyophilized exosomes derived from adipose tissue (ADSCs) and Wharton's jelly (WJSCs) mesenchymal stem cells, administered via spray and injection methods, on chronic wound healing in a rat model of spinal cord injury. Adipose tissue and Wharton's jelly samples were collected from healthy donors during liposuction and cesarean section procedures. Mesenchymal stem cells were isolated and characterized by flow cytometry for CD29, CD90, CD105 (positive) and CD34, CD45 (negative markers). Exosomes were extracted by ultracentrifugation, lyophilized, and characterized by scanning electron microscopy (SEM). Growth factors (PDGF and FGF) were measured by ELISA. In vivo, 15 adult male Wistar rats were divided into five groups (n=3): control (normal saline), L-AEXO i (lyophilized adipose-derived exosomes, injection), L-WEXO i (lyophilized Wharton's jelly-derived exosomes, injection), L-AEXO l (lyophilized adipose-derived exosomes, spray), and L-WEXO l (lyophilized Wharton's jelly-derived exosomes, spray). Full-thickness skin wounds (1×1 cm) were created on the dorsum under general anesthesia. Treatments were administered five days post-wounding. After 21 days, biopsies were collected for histopathological evaluation (H&E and Masson's trichrome), immunohistochemical staining (Collagen I), and quantitative gene expression analysis (Collagen I, Fibronectin, Elastin) by real-time PCR. Results demonstrated significantly increased cell viability in all treatment groups compared to control (P<0.05). The L-WEXO i group showed the highest expression of Collagen I (3.5-fold), Fibronectin (3.5-fold), and Elastin (4.5-fold) compared to control (P<0.001). Histological examination revealed superior epithelialization, collagen deposition, and reduced inflammation in treatment groups, particularly in L-WEXO i and L-WEXO l groups. Lyophilized exosomes derived from Wharton's jelly mesenchymal stem cells, administered via both injection and spray methods, significantly enhance chronic wound healing in a rat model of spinal cord injury.