The utilization of mesenchymal stem cell (MSC)-derived exosomes, which include numerous growth factors, cytokines, and microRNAs, is the primary aspect of the novel MSC activity models. The current research aims to: (i) identify the morphology of exosomes; (ii) determine exosomes secreted into MSCs conditioned cell culture medium; and (iii) perform a comprehensive characterization of isolated exosomes and elucidate their protective role in the diabetic nephropathy animal model. Ultracentrifugation was performed by utilizing the culture supernatant of MSCs. Transmission electron microscopy, nanoparticle tracking analysis, as well as Western blot, were utilized for isolated exosome characterization. The purified exosomes were used for in vivo implantation in a diabetic nephropathy animal model. The present research was carried out on 70 adult male albino rats weighing 180 to 200 grams. Rats were classified into seven groups: Group I: negative control group; Group II: diabetic nephropathy group; Group III: Balanites therapeutic group; Group IV: Balanites + MSCs therapeutic group; Group V: Balanites + exosome therapeutic group; Group VI: MSCs therapeutic group; and Group VII: exosome therapeutic group. By the end of the study period, total antioxidant capacity (TAC), malondialdehyde (MDA), and the histology of pancreatic tissue were assessed. Isolated exosomes with sizes ranging from 30 to 150 nm demonstrated the typical cup-shaped morphology. Additionally, exosome criteria were demonstrated by the exosome surface proteins CD81 and CD63, which were expressed by exosome marker proteins. Treatment with exosomes along with Balanites induced a significant reduction in pancreatic MDA with a substantial elevation in pancreatic TAC. Furthermore, treatment with exosomes and Balanites demonstrated normal pancreatic parenchyma and pancreatic lobules with normal pancreatic acini and acinar cells. These findings strongly suggest that ultracentrifugation is the most efficient tool for isolating exosomes. Also, these findings demonstrated that Balanites and exosomes had synergistic effects on one another, with more potent renoprotective activities in rats.