Gentamicin-Loaded Exosomes from IMMUNEPOTENT CRP Enhance Healing of Infected Diabetic Wound in Mice

Paola Leonor García-Coronado¹Brandón Alberto Garza Martínez¹Kenia Arisbe Moreno-Amador¹David Reding¹Diana Ginette Zarate Triviño¹Diana Caballero-Hernández¹Pablo Zapata Benavides¹Gabriel Luna-Barcenas^2*Cristina Rodríguez-Padilla¹Moisés Armides Franco Molina^1*

• ¹Facultad de Ciencias Biológicas. Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Mexico
• ²School of Engineering and Sciences, The Institute of Advanced Materials for Sustainable Manufacturing, Tecnológico de Monterrey, Queretaro, Mexico

Diabetic foot infections (DFIs) are a major cause of lower extremity amputations and are associated with substantial morbidity and reduced quality of life. Given the limited efficacy of current treatments and the rise of antimicrobial resistance, there is an urgent need for innovative therapeutic approaches. This study evaluates the use of exosomes derived from a bovine leukocyte spleen extract (IMMUNEPOTENT CRP), loaded with gentamicin, to improve infection control and promote wound healing in a diabetic setting. A full-thickness wound model was established in streptozotocin (STZ)-induced diabetic mice, followed by inoculation with Staphylococcus aureus to simulate infected diabetic ulcers. Mice were treated with gentamicin-loaded exosomes (Exo-Genta), IMMUNEPOTENT CRP-derived exosomes (ICRP-Exo), or free gentamicin. Exo-Genta efficiently released gentamicin under both acidic and basic conditions, exhibited prolonged release under alkaline conditions. Notably, the Exo-Genta group exhibited significantly faster wound closure by day 9 (p < 0.0001) and a decreased bacterial load (49 folds-98%); p < 0.001) compared to all other groups. Histological analyses revealed that treatments involving Exo-Genta, ICRP-Exo, and IMMUNEPOTENT CRP significantly enhanced collagen fiber deposition (p < 0.05), blood vessel formation, and hair follicle regeneration. At the molecular level, these treatments increased AKT phosphorylation (p < 0.05) and modulated the inflammatory response, with reduced levels of TNF-α, IL-6, and MCP-1 (all p < 0.05), alongside a significant increase in anti-inflammatory IL-10 (p < 0.05). In conclusion, gentamicin-loaded exosomes derived from IMMUNEPOTENT CRP demonstrated enhanced antimicrobial activity and tissue regeneration in infected diabetic wounds. These findings support their potential as an effective and less invasive therapeutic strategy for diabetic foot ulcers by combining infection control with pro-regenerative and immunomodulatory effects.