The Therapeutic Potential of Small Extracellular Vesicles Derived From Cucurbita pepo in Wound Healing: A Regenerative Medicine Perspective

Wound healing is a multifaceted biological process involving cellular, biochemical, and physiological events regulated by cytokines and growth factors. Recent advancements in regenerative medicine have highlighted the therapeutic potential of plant-derived small extracellular vesicles (SEVs). This study investigates the wound-healing properties of SEVs isolated from Cucurbita pepo (zucchini) juice. Using an aqueous two-phase system, homogenous SEVs were isolated and characterized through scanning electron microscopy (SEM), flow cytometry, and nanoparticle tracking analysis. The SEVs displayed exosome-specific markers, including CD9, CD63, CD81 and HSP70, and exhibited a size range of 50-150 nm. In vitro assays revealed that zucchini-derived SEVs significantly enhance cellular processes critical to wound healing. A 200 µg/mL concentration of SEVs increased cell viability and proliferation in human dermal fibroblasts (HDF), human keratinocytes (HaCaT), and human umbilical vein endothelial cells (HUVEC) over 24, 48, and 72 hours. Scratch assays demonstrated accelerated cell migration, achieving near-complete wound closure within 48 hours. Furthermore, SEVs mitigated oxidative stress in H₂O₂-induced inflammation models, improving cell viability and showcasing anti-inflammatory effects. Notably, quantitative RT-PCR analysis showed a 30% upregulation in collagen type I gene expression, underscoring the SEVs' role in extracellular matrix remodeling and tissue regeneration. The findings indicate that Cucurbita pepo SEVs possess multifunctional properties—stimulating cell proliferation, migration, anti-inflammatory activity, and collagen synthesis—making them a promising therapeutic agent in wound healing. This study establishes a foundation for further exploration of plant-derived SEVs in regenerative medicine, particularly for tissue repair and wound management.