Establishing the long-term alcohol model.

<p>(<b>A</b>) A schematic diagram depicting the long-term alcohol exposure model used in these studies is shown<b>.</b> Caco-2 cells were treated with 0.2% ethanol or 0.2% H₂O (control) daily for 2 months. Confluent monolayers were either kept intact (uninjured) or scrape-wounded as described in materials and methods (Wound model). (<b>B</b>) Control and EtOH Caco-2 confluent monolayers were scrape wounded. The impact of long-term ethanol exposure on the healing dynamics of Caco-2 wounded epithelium was measured by time-lapse videomicroscopy. Representative frames at indicated time points post wounding are shown. (<b>C</b>) Wound areas were measured and the tabulated results relative to the original wound size, indicating that EtOH treated cells heal faster than the control. (<b>D & E</b>) The cytotoxic impact of long-term alcohol exposure on Caco-2 confluent monolayer was evaluated by IF time-lapse videomicroscopy, using PI uptake (red cells are dead) as described in materials and methods. Representative frames at indicated time points are shown in (D) and the results were tabulated from each frame and are shown in (E), indicating that EtOH treatment does not render Caco-2 cells prone to enhanced cytotoxicity. Data were plotted as the Mean ± SEM (* indicates significance with <i>p</i><0.05, n = 3). Arrows point to cluster of cells moving inward.