Bruce J. Walters

Determinants of Plasma Membrane Wounding by Deforming Stress

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.

Contractile response to colonic distention is influenced by oscillation frequency.

Abstract  Static colonic mechanical properties are characterized by stepwise balloon distention. It is unclear whether the state of contractile activation affects frequency‐dependent differences in biomechanical properties. Our aim was to investigate the frequency‐dependence of colonic mechanical properties by sinusoidal oscillation. A descending colonic balloon was sinusoidally oscillated by 25 mL at 5, 10 and 20 cpm in randomized order for 20 min at each frequency in six healthy subjects before and after neostigmine. Volume oscillation was between 75–100 mL before, and 25–50 mL after neostigmine. Pressure waveforms were most variable shortly after commencing oscillation, reflecting an initial contractile response to distention. Elastance (i.e. pressure response to imposed volume) and hysteresivity were estimated; hysteresivity represents the proportion of energy added to the system during inflation, which cannot be recovered during deflation. Colonic elastance was frequency dependent, being highest and most variable at 10 cpm. In contrast, hysteresivity was not significantly different across frequencies. Neostigmine increased mean colonic elastance at all frequencies, and hysteresivity only at 5 cpm. Thus, colonic mechanical properties, particularly elastance are frequency‐dependent. The frequency‐dependence of colonic mechanical properties is worthy of future study because it may provide insights into reflex responses in health and disease.