4.3. Endothelial Cell Migration Assay

A wound-healing assay was used to determine the migratory ability of endothelial cells [29]. Briefly, HUEVCs, at a density of 2 × 10⁵ cells/well, were seeded into each well of a sterile 12-well plate. After cells incubated for 24 h, with application of a sterile 200 μL plastic pipette tip, a single and straight wound was manually made at the center of cell monolayer. To remove scratched cells, prewarmed PBS was used to wash cell. Images were taken under a phase-contrast microscope (A_t0). After cells incubated with complete medium containing VEGF (10 ng/mL) or Avastin/piceatannol for 8 h, four wound fields were chosen randomly and photographed (A_t8). By using Tscratch software (CSE Lab, Switzerland), the scratched area of each group before and after drug treatments was determined and based on the below formula: Recovery (%) = A_t0 − A_t8/A_t0 × 100%, the recovery percentage of cells was calculated.