Wound healing
When the epithelium was fully differentiated, hydrocortisone was
withdrawn from the basolateral media for three days prior to wounding.
Mechanical injury was performed by scraping a P10 sterile pipette tip
across the cell layer creating a wound with a diameter ranging from 600
to 1500μm. The initial wound area was expressed as 100% to remove
variability from wounds of different size 6. Wounds
were considered to be closed when the calculated area fell below 3%,
the effective limit of detection for image processing. Wound closure was
calculated as follows: Wound closure (%) = 100 - ((Area/Initial
Area)*100). Wound closure (%), plotted as a function of time (hour),
was used to calculate the rate of wound closure (%/hour).