Recent advances in the ventilation of patients with the Acute Respiratory Distress Syndrome (ARDS), including ventilation at low lung volumes, have resulted in a decreased mortality rate. However, even low lung volume ventilation may exacerbate lung injury due to the cyclic opening and closing of fluid-occluded airways. Specifically, the hydrodynamic stresses generated during airway reopening may result in epithelial cell (EpC) injury. We utilized an in-vitro cell-culture model of airway reopening to investigate the effect of reopening velocity, airway diameter, cell confluence and cyclic closure/reopening on cellular injury. Reopening dynamics were simulated by propagating a constant velocity air bubble in an adjustable-height, parallel-plate flow chamber. This chamber was occluded with different types of fluids and contained either a confluent or sub-confluent monolayer of EpC. Fluorescent microscopy was used to quantify morphological properties and the percentage of dead cells under different experimental conditions. Decreasing the channel height and reopening velocity resulted in a larger percentage of dead cells due to an increase in the spatial pressure gradient applied to the EpC. These results indicate that distal regions of the lung are more prone to injury and that rapid inflation may be cyto-protective. Repeated reopening events and sub-confluent conditions resulted in significant cellular detachment. In addition, we observed a larger percentage of dead cells under sub-confluent conditions. Analysis of this data suggests that in addition to the magnitude of the hydrodynamic stresses generated during reopening, the EpC's morphological, biomechanical and micro-structural properties may also be important determinants of cell injury.