Targeting Fibrotic Scars with Extracellular Vesicles Extracted from Mature Aloe vera: Enhanced Antioxidant, Anti-inflammatory, and Antifibrotic Activity through M2 Macrophage Polarization and Myofibroblasts Inhibition

BackgroundAbnormal scarring and fibrotic skin disorders arise from dysregulated wound healing processes. While Aloe vera is widely recognized for its therapeutic properties, the potential of its extracellular vesicles (Av-EVs) remains underexplored. ObjectiveThis study aimed to isolate and characterize Av-EVs and evaluate their antioxidant, anti-inflammatory, and antifibrotic properties in vitro, focusing on the impact of extraction method and plant maturity. MethodsAv-EVs were isolated from mature and young Aloe vera leaves using manual (NB) or blender-based (B) homogenization. Vesicles were characterized by nanoparticle tracking analysis, transmission electron microscopy, and protein quantification. Antioxidant and cytotoxicity assays (DPPH, alamarBlue) were followed by functional anti-inflammatory and antifibrotic analyses respectively in LPS-stimulated THP-1 macrophages and TGF-{beta}1/Vitamin C-activated human dermal fibroblasts (RT-qPCR, immunofluorescence, proteomics). ResultsNB-derived EVs from mature leaves exhibited the most potent activity across all assays, showing superior antioxidant capacity, greater suppression of pro-inflammatory cytokines, enhanced M2 macrophage polarization, and significant downregulation of COL1A1 and -SMA. In contrast, B-derived and young leaf-derived EVs showed reduced bioactivity, with young EVs failing to inhibit fibrotic markers. ConclusionManually extracted Av-EVs from mature leaves demonstrate superior multifunctional bioactivity, highlighting their potential as plant-derived nanotherapeutics for fibrotic scar modulation.