Influence of cytoskeletal structure and mechanics on epithelial cell injury during cyclic airway reopening

Yalcin, Huseyin C.; Hallow, K. Melissa; Wang, J.; Wei, Ming-Tzo; Ou-Yang, H. Daniel; Ghadiali, Samir N.

doi:10.1152/ajplung.90562.2008

Cited by 46 publications

(73 citation statements)

References 43 publications

(58 reference statements)

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“…This could fundamentally alter the force transmission and the effects of deforming stress on the PM-CSK interface. This may also explain why some authors have proposed that interventions that decrease cell stiffness may be cytoprotective (26).…”

Section: Discussionmentioning

confidence: 99%

Determinants of plasma membrane wounding by deforming stress

Oeckler¹,

Lee

Park

et al. 2010

American Journal of Physiology-Lung Cellular and Molecular Phys

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Once excess liquid gains access to air spaces of an injured lung, the act of breathing creates and destroys foam and thereby contributes to the wounding of epithelial cells by interfacial stress. Since cells are not elastic continua, but rather complex network structures composed of solid as well as liquid elements, we hypothesize that plasma membrane (PM) wounding is preceded by a phase separation, which results in blebbing. We postulate that interventions such as a hypertonic treatment increase adhesive PM-cytoskeletal (CSK) interactions, thereby preventing blebbing as well as PM wounds. We formed PM tethers in alveolar epithelial cells and fibroblasts and measured their retractive force as readout of PM-CSK adhesive interactions using optical tweezers. A 50-mOsm increase in media osmolarity consistently increased the tether retractive force in epithelial cells but lowered it in fibroblasts. The osmo-response was abolished by pretreatment with latrunculin, cytochalasin D, and calcium chelation. Epithelial cells and fibroblasts were exposed to interfacial stress in a microchannel, and the fraction of wounded cells were measured. Interventions that increased PM-CSK adhesive interactions prevented blebbing and were cytoprotective regardless of cell type. Finally, we exposed ex vivo perfused rat lungs to injurious mechanical ventilation and showed that hypertonic conditioning reduced the number of wounded subpleural alveolus resident cells to baseline levels. Our observations support the hypothesis that PM-CSK adhesive interactions are important determinants of the cellular response to deforming stress and pave the way for preclinical efficacy trials of hypertonic treatment in experimental models of acute lung injury. alveolar epithelial cell; acute lung injury; interfacial stress; osmotic response; cytoprotection THE SYNDROME OF ventilator-induced lung injury (VILI) contributes to the morbidity and mortality of critically ill patients (25). The clinical manifestations of the syndrome are indistinguishable from those of all-cause acute lung injury and at their core reflect a mechanotransduction event, i.e., the effects of deforming stress on cell and tissue injury, remodeling, and repair. The complex topographical distributions of lung mechanical properties, and hence of parenchymal stress and strain together with the numerous and nuanced injury manifestations, make it very difficult to establish mechanistic cause and effect relationships on the scale of interest, e.g., that of individual cells. Our research, therefore, focuses on a very specific mechanotransduction event, namely physical stress-related epithelial cell wounding and repair, which we believe contributes to the pathogenesis VILI.In the current study, we present a body of work in which we have explored the effects of hypertonic exposure on cell wounding in experimental models ranging from individual cells to rodent VILI preparations. The experiments were not designed to test the preclinical efficacy of an intervention, but rather to explore a cell bio...

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Section: Discussionmentioning

confidence: 99%

Determinants of plasma membrane wounding by deforming stress

Oeckler¹,

Lee

Park

et al. 2010

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Briefly, substrates with cultured A549 cells or HSAEpC were placed inside the FCS2 chamber, and silicone gaskets were used to create a parallel plate flow channel (35-mm length, 10-mm width, and 0.5-mm height). Similar to previous studies (47,48), this channel was then filled with a surfactant-deficient PBS fluid using a programmable PHD 2000 syringe pump (Harvard Apparatus, Holliston, MA). The fluid was then retracted from the channel at a flow rate (0.09 ml/min) to create an air bubble that propagated over the cell monolayer at a velocity of 0.3 mm/s (Fig.…”

Section: Fluid-filled Airway Reopening Simulationmentioning

confidence: 99%

“…Under these conditions, the closure and reopening of fluid-filled airways generates interfacial and microbubble flows that can significantly damage the lung epithelium. Specifically, several investigators have used computational (13,14,23,25,29) and in vitro experimental (3,30,47,48) techniques to demonstrate that this interfacial flow generates complex mechanical forces that cause cell necrosis/plasma membrane disruption and detachment of cells from their substrate. In addition, mechanical forces associate with atelectrauma can also activate inflammatory signaling pathways (27,28).…”

mentioning

confidence: 99%

“…These authors demonstrated that the large pressure gradient near the bubble front causes plasma membrane disruption and cell necrosis (3,30). Using a similar rigid airway model, Yalcin et al (48) demonstrated that the degree of cell injury during cyclic airway closure/reopening is inversely proportional to airway diameter, and that changes in the cell's cytoskeletal structure can be used to mitigate cell injury during reopening (47). Similarly, Oeckler et al (35) demonstrated that hypertonic treatment reduces cell injury by increasing plasma membrane-cytoskeleton adhesive interactions.…”

mentioning

confidence: 99%

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Influence of airway wall compliance on epithelial cell injury and adhesion during interfacial flows

Higuita‐Castro

Mihai

Hansford

et al. 2014

Journal of Applied Physiology

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Higuita-Castro N, Mihai C, Hansford DJ, Ghadiali SN. Influence of airway wall compliance on epithelial cell injury and adhesion during interfacial flows. J Appl Physiol 117: 1231-1242, 2014. First published September 11, 2014 doi:10.1152/japplphysiol.00752.2013.-Interfacial flows during cyclic airway reopening are an important source of ventilator-induced lung injury. However, it is not known how changes in airway wall compliance influence cell injury during airway reopening. We used an in vitro model of airway reopening in a compliant microchannel to investigate how airway wall stiffness influences epithelial cell injury. Epithelial cells were grown on gel substrates with different rigidities, and cellular responses to substrate stiffness were evaluated in terms of metabolic activity, mechanics, morphology, and adhesion. Repeated microbubble propagations were used to simulate cyclic airway reopening, and cell injury and detachment were quantified via live/dead staining. Although cells cultured on softer gels exhibited a reduced elastic modulus, these cells experienced less plasma membrane rupture/necrosis. Cells on rigid gels exhibited a minor, but statistically significant, increase in the power law exponent and also exhibited a significantly larger height-to-length aspect ratio. Previous studies indicate that this change in morphology amplifies interfacial stresses and, therefore, correlates with the increased necrosis observed during airway reopening. Although cells cultured on stiff substrates exhibited more plasma membrane rupture, these cells experienced significantly less detachment and monolayer disruption during airway reopening. Western blotting and immunofluorescence indicate that this protection from detachment and monolayer disruption correlates with increased focal adhesion kinase and phosphorylated paxillin expression. Therefore, changes in cell morphology and focal adhesion structure may govern injury responses during compliant airway reopening. In addition, these results indicate that changes in airway compliance, as occurs during fibrosis or emphysema, may significantly influence cell injury during mechanical ventilation.ventilation-induced lung injury; airway compliance; airway reopening; cell mechanics; and cell injury THE ACUTE RESPIRATORY DISTRESS syndrome (ARDS) is a lifethreatening condition that involves disruption of the alveolarcapillary barrier and impaired gas exchange (46).1 Although mechanical ventilation of ARDS patients can restore normal oxygenation, these ventilators may also cause significant cellular injury and trigger proinflammatory signaling cascades (26, 31) in a process known as ventilator-induced lung injury (VILI). One form of VILI, known as volutrauma, involves the overdistension and cyclic stretching of lung tissue, which causes epithelial cell necrosis (43), disruption of the alveolarcapillary barrier (7), and inflammatory signaling (44). Presently, protective ventilation strategies, such as low tidal volumes, are the standard of care and seek to minimize volutrau...

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“…Several studies suggest that excessive mechanical stretch of lung epithelial cells can cause direct injury involving damage to cells 11,13,26,27 . This type of damage is difficult to capture without real-time imaging during the mechanical distention.…”

Section: Direct Epithelial Monolayer Damage Due To Stretchmentioning

confidence: 99%

Live Cell Imaging during Mechanical Stretch

Rapalo

Herwig

Hewitt

et al. 2015

JoVE

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There is currently a significant interest in understanding how cells and tissues respond to mechanical stimuli, but current approaches are limited in their capability for measuring responses in real time in live cells or viable tissue. A protocol was developed with the use of a cell actuator to distend live cells grown on or tissues attached to an elastic substrate while imaging with confocal and atomic force microscopy (AFM). Preliminary studies show that tonic stretching of human bronchial epithelial cells caused a significant increase in the production of mitochondrial superoxide. Moreover, using this protocol, alveolar epithelial cells were stretched and imaged, which showed direct damage to the epithelial cells by overdistention simulating one form of lung injury in vitro. A protocol to conduct AFM nano-indentation on stretched cells is also provided. Video LinkThe video component of this article can be found at

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Influence of cytoskeletal structure and mechanics on epithelial cell injury during cyclic airway reopening

Cited by 46 publications

References 43 publications

Determinants of plasma membrane wounding by deforming stress

Determinants of plasma membrane wounding by deforming stress

Influence of airway wall compliance on epithelial cell injury and adhesion during interfacial flows

Live Cell Imaging during Mechanical Stretch

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