Angiogenic Response to Atorvastatin and Human Umbilical Cord Blood-Mesenchymal Stem Cell-Derived Exosomes in the IN-OVO Chorioallantoic Membrane Model

Abstract

Exosomes derived from human umbilical cord blood–mesenchymal stem cells (hUCB-MSCs) contain various proangiogenic mediators, whereas atorvastatin exerts pleiotropic endothelial effects through the PI3K/Akt/eNOS pathway and enhanced nitric oxide bioavailability. However, the angiogenic responses to atorvastatin, hUCB-MSC-derived exosomes, and their combination have not been fully elucidated.. To compare the angiogenic effects of atorvastatin, hUCB-MSC-derived exosomes, and their combination in an in ovo chorioallantoic membrane (CAM) model. Four groups were included: control, atorvastatin, hUCB-MSC-derived exosome, and exosome plus atorvastatin. Seven embryonated chicken eggs per group were assessed at D0, D1, and D2, with vascular density as the primary outcome. A significant time-by-group interaction was observed for vascular density, F(6,48) = 23.278, p < 0.001, partial η² = 0.744. Cumulative vascular density change from D0 to D2 differed significantly among the control, atorvastatin, exosome, and combination groups (0.671 ± 0.647, 3.196 ± 0.734, 4.786 ± 0.828, and 2.642 ± 0.683 percentage points, respectively; F(3,24) = 38.302, p < 0.001, η² = 0.827). Exosomes produced greater increases than control (p < 0.001), atorvastatin (p = 0.002), and combination treatment (p < 0.001). Atorvastatin and combination treatment also exceeded control (both p < 0.001), whereas they did not differ from each other (p = 0.495). Atorvastatin and hUCB-MSC-derived exosomes induced significant proangiogenic responses in the in ovo CAM model. hUCB-MSC-derived exosomes produced the strongest angiogenic response. hUCB-MSC-derived exosomes produced the strongest proangiogenic response, while atorvastatin also increased vascular density. Adding atorvastatin to exosomes provided no additional angiogenic benefit at the doses evaluated.