A Multiomic Study of Platelet-Derived Extracellular Vesicles and Impact of Platelet Concentrate Sources.

Platelet-derived extracellular vesicles (pEVs) are a potent fraction of platelet concentrates, enhancing their therapeutic potential in regenerative medicine. This study evaluates pEV from three platelet sources: platelet lysate (PL), fresh platelets (fPs), and aged platelets (aPs), to determine how activation and storage conditions affect pEV characteristics, functionality, and molecular content. pEV are isolated using size exclusion chromatography (SEC) and characterized by transmission electron microscopy (TEM), western blot, and nanoparticle tracking analysis (NTA). Functional assays include wound healing, metabolic activity, and cytotoxicity. Protein and miRNA profiles are obtained through LC-MS/MS and miRNA arrays, followed by bioinformatic analysis. Findings show that PL-derived pEV exhibits the highest yield and purity, containing markers CD63 and CD9. Enhanced fibroblast migration in wound healing assays suggest a critical role for PL-pEV in hemostasis, proliferation, and remodeling phases. Multiomics analysis identifies upregulated miRNAs, particularly miR-210-3p and the miR-320 family, associated with wound healing. Differential protein analysis reveals an enrichment in immune response and wound healing pathways within PL-pEV. These results demonstrate the impact of platelet preparation methods on pEV molecular cargo and efficacy, with hsa-miR-320a, hsa-miR-320b, and hsa-miR-210-3p identified as key mediators supporting the clinical potential of PL-pEV in regenerative medicine.