Abstract

Background: The cosmetic industry is a rapidly expanding, high-value sector that drives continuous research into biologically active compounds and advanced delivery technologies aimed at improving skin health. Objective: To investigate the biological activity of apple-derived extracellular vesicles (ADEVs) and to evaluate strategies to enhance the transcutaneous penetration of ADEV-based formulations. Methods: In vitro, purified ADEVs were characterized in terms of size distribution and concentration. Their internalization was assessed in different cell types, and gene expression analysis was performed in treated cellular models. Ex vivo, PKH26-labeled ADEVs were applied to human surgical skin samples, either alone or in combination with microneedling (MN) using different procedural protocols. Fluorescence distribution was quantified in both epidermal and dermal compartments. Results: ADEV characterization showed a size range of 102–160 nm and a particle concentration of 9.48 × 1011 particles/mL, together with morphological features consistent with EVs. ADEVs demonstrated efficient cellular internalization in vitro and modulated the expression of selected target genes (CDH5, NOS3, KLF2, and KLF4). In the ex vivo model, fluorescence signal associated with PKH26-labeled ADEV-based formulations was detected within human skin layers. A significant treatment effect was observed in both epidermal (F = 33.57, p < 0.001) and dermal compartments (F = 7.57, p = 0.018), with the highest fluorescence signal consistently detected when EV application was preceded by MN (MN + EVs). Notably, ADEV-based formulation alone also induced a significant increase in the epidermis compared with untreated controls, although of lower magnitude than that observed following MN pre-treatment. Conclusions: These findings support a controlled evaluation of microneedling as a strategy to enhance the distribution of ADEV-based formulations within human skin, highlighting the importance of application sequence. More broadly, this work contributes to the development of standardized and reproducible delivery approaches, while underscoring the need for further methodological refinement to achieve vesicle-specific tracking in complex tissues.

IPC Classification

Keywords