2.7. Scratch wound assay.

Cells (2.5×10⁵ cells/well) were grown in 6-well plates with complete medium until confluence and placed in serum-free medium for 24 h, then a wound in the cell monolayer was performed with a sterile pipette tip. After washing with PBS, RPMI was refreshed, and drags were added. Pictures at initial time and after 16 h of incubation were collected. Image analysis included the assessment of the initial and final areas of the wound using the ImageJ 1.51j8. Results are expressed as percentage of migration according to the formula 100-((final area*100)/initial area). Three wounds were made in each well and 9 to 12 microphotographs were taken for each condition.