Microfluidic shear stress-regulated surfactant secretion in alveolar epithelial type II cells in vitro.
We investigated the role of flow-induced shear stress on the mechanisms regulating surfactant secretion in type II alveolar epithelial cells (ATII) using microfluidic models. Following flow stimulation spanning a range of wall shear stress (WSS) magnitudes, monolayers of ATII (MLE-12 and A549) cells were examined for surfactant secretion by evaluating essential steps of the process, including relative changes in the number of fusion events of lamellar bodies (LBs) with the plasma membrane (PM) and intracellular redistribution of LBs. F-actin cytoskeleton and calcium levels were analyzed in A549 cells subjected to WSS spanning 4-20 dyn/cm(2). Results reveal an enhancement in LB fusion events with the PM in MLE-12 cells upon flow stimulation, whereas A549 cells exhibit no foreseeable changes in the monitored number of fusion events for WSS levels ranging up to a threshold of ∼8 dyn/cm(2); above this threshold, we witness instead a decrease in LB fusion events in A549 cells. However, patterns of LB redistribution suggest that WSS can potentially serve as a stimulus for A549 cells to trigger the intracellular transport of LBs toward the cell periphery. This observation is accompanied by a fragmentation of F-actin, indicating that disorganization of the F-actin cytoskeleton might act as a limiting factor for LB fusion events. Moreover, we note a rise in cytosolic calcium ([Ca(2+)]c) levels following stimulation of A549 cells with WSS magnitudes ranging near or above the experimental threshold. Overall, WSS stimulation can influence key components of molecular machinery for regulated surfactant secretion in ATII cells in vitro.
Technion - Israel Institute of Technology, Haifa, Israel. sznitman@bm.technion.ac.il.
Sanjeev Kumar
SK
Mahto
Janna
J
Tenenbaum-Katan
Ayala
A
Greenblum
Barbara
B
Rothen-Rutishauser
Josué
J
Sznitman
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism*; D005089:Exocytosis; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D044085:Microfluidics; D015388:Organelles / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000378:metabolism*; D013312:Stress, Physiological
D011663:Pulmonary Surfactants; D002118:Calcium
F-actin cytoskeleton; alveolar epithelial cells; lamellar bodies; microfluidics; pulmonary surfactant; shear stress