3×EV treatment of discs lead to increased transcription of fibrillin-1 after 30 days of culture. Fibrin discs were cultured for 30 days using treatments replenished every 48–72 h, subjected to RNA extraction, and analyzed by RT-qPCR for expression of human (A) tropoelastin, (B) fi-brillin-1, (C) fibulin-4, (D) fibulin-5, (E) lysyl oxidase (LOX), (F) lysyl oxidase-like 1 (LOXL-1), and (G) latent TGF-β binding protein-4 (LTBP-4). Treatment groups were non-conditioned media (nCM), ASC conditioned media (CM), CM depleted of EVs (dCM), 1× extracellular vesicle treatment (EV×1), or 3×EV treatment (EV×3). Gene expression was expressed as ΔCt relative to the housekeeping gene GAPDH, where higher values relative to nCM indicate higher expression levels. For all groups, the individual data points are plotted as dots, with a line indicating the average value. * = p < 0.05 relative to nCM.