Acoustically detectable cellular-level lung injury induced by fluid mechanical stresses in microfluidic airway systems

We describe a microfabricated airway system integrated with computerized air–liquid two-phase microfluidics that enables on-chip engineering of human airway epithelia and precise reproduction of physiologic or pathologic liquid plug flows found in the respiratory system. Using this device, we demonstrate cellular-level lung injury under flow conditions that cause symptoms characteristic of a wide range of pulmonary diseases. Specifically, propagation and rupture of liquid plugs that simulate surfactant-deficient reopening of closed airways lead to significant injury of small airway epithelial cells by generating deleterious fluid mechanical stresses. We also show that the explosive pressure waves produced by plug rupture enable detection of the mechanical cellular injury as crackling sounds.

[1]  Yang Pc,et al.  (Am. J. Respir. Cell Mol. Biol., 32:540-547)Autocrine and Paracrine Regulation of IL-8 Expression in Lung Cancer Cells , 2005 .

[2]  R D Kamm,et al.  Mechanical stress is communicated between different cell types to elicit matrix remodeling , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Gaver,et al.  The influence of surfactant on two-phase flow in a flexible-walled channel under bulk equilibrium conditions , 1998 .

[4]  Donald P. Gaver,et al.  The steady propagation of a bubble in a flexible-walled channel: Asymptotic and computational models , 2002 .

[5]  S. Margulies,et al.  Equibiaxial deformation-induced injury of alveolar epithelial cells in vitro. , 1998, American journal of physiology. Lung cellular and molecular physiology.

[6]  Melanie M. Choe,et al.  Extracellular matrix remodeling by dynamic strain in a three-dimensional tissue-engineered human airway wall model. , 2006, American journal of respiratory cell and molecular biology.

[7]  H. Wirtz,et al.  Calcium mobilization and exocytosis after one mechanical stretch of lung epithelial cells. , 1990, Science.

[8]  P. Paré,et al.  The nature of small-airway obstruction in chronic obstructive pulmonary disease. , 2004, The New England journal of medicine.

[9]  C. Waters,et al.  Mechanical strain inhibits repair of airway epithelium in vitro. , 1998, American journal of physiology. Lung cellular and molecular physiology.

[10]  N. Krug,et al.  Dysfunction of pulmonary surfactant in asthmatics after segmental allergen challenge. , 1999, American Journal of Respiratory and Critical Care Medicine.

[11]  M. Ballmann,et al.  Pulmonary surfactant, lung function, and endobronchial inflammation in cystic fibrosis. , 2004, American journal of respiratory and critical care medicine.

[12]  P. Piirilä,et al.  Crackles: recording, analysis and clinical significance. , 1995, The European respiratory journal.

[13]  Yu Wang,et al.  Mechanical stretch promotes alveolar epithelial type II cell differentiation. , 2001, Journal of applied physiology.

[14]  S. Brody,et al.  Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[15]  E. Weibel,et al.  Architecture of the Human Lung , 1962, Science.

[16]  Shuichi Takayama,et al.  Reversible switching of high-speed air-liquid two-phase flows using electrowetting-assisted flow-pattern change. , 2003, Journal of the American Chemical Society.

[17]  D. Gaver,et al.  An investigation of pulmonary surfactant physicochemical behavior under airway reopening conditions. , 2000, Journal of applied physiology.

[18]  M. Post,et al.  Apoptosis in Lung Development and Neonatal Lung Injury , 2004, Pediatric Research.

[19]  R. Tirouvanziam,et al.  Primary inflammation in human cystic fibrosis small airways. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[20]  A. W. Murray,et al.  Cyclic stretch induces both apoptosis and secretion in rat alveolar type II cells , 1999, FEBS letters.

[21]  S. Russo,et al.  The late asthmatic response is linked with increased surface tension and reduced surfactant protein B in mice. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[22]  Donald P. Gaver,et al.  The steady motion of a semi-infinite bubble through a flexible-walled channel , 1996, Journal of Fluid Mechanics.

[23]  Hideki Fujioka,et al.  Steady propagation of a liquid plug in a two-dimensional channel. , 2004, Journal of biomechanical engineering.

[24]  H. Pasterkamp,et al.  Respiratory sounds. Advances beyond the stethoscope. , 1997, American journal of respiratory and critical care medicine.

[25]  Unsteady bubble propagation in a flexible channel: predictions of a viscous stick-slip instability , 2005 .

[26]  P. Dargaville,et al.  Surfactant abnormalities in infants with severe viral bronchiolitis. , 1996, Archives of disease in childhood.

[27]  H. Stanley,et al.  Crackles and instabilities during lung inflation , 2005 .

[28]  D. Dreyfuss,et al.  Ventilator-induced lung injury: lessons from experimental studies. , 1998, American journal of respiratory and critical care medicine.

[29]  S. Randell,et al.  A primary culture model of differentiated murine tracheal epithelium. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[30]  J. E. Anderson,et al.  Influence of strain on [3H]thymidine incorporation, surfactant-related phospholipid synthesis, and cAMP levels in fetal type II alveolar cells. , 1993, American journal of respiratory cell and molecular biology.

[31]  J. Hohlfeld.  The role of surfactant in asthma , 2001, Respiratory research.

[32]  Arthur S Slutsky,et al.  Tidal ventilation at low airway pressures can augment lung injury. , 1994, American journal of respiratory and critical care medicine.

[33]  Kay C Dee,et al.  Mechanisms of surface-tension-induced epithelial cell damage in a model of pulmonary airway reopening. , 2003, Journal of applied physiology.

[34]  马建新,et al.  用FEV6．0代替FVC诊断气道阻塞和肺功能受限[英]／Swanney MP…∥Am J Respir Crit Care Med． , 2002 .

[35]  Melanie M. Choe,et al.  An in vitro airway wall model of remodeling. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[36]  T. Nakamura,et al.  Mechanical strain and dexamethasone selectively increase surfactant protein C and tropoelastin gene expression. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[37]  D. Gaver,et al.  An investigation of the influence of cell topography on epithelial mechanical stresses during pulmonary airway reopening. , 2005, Physics of fluids.

[38]  J. Sznajder,et al.  Mechanical stretching of alveolar epithelial cells increases Na(+)-K(+)-ATPase activity. , 1999, Journal of applied physiology.

[39]  J. Koo,et al.  Regulation of the secretory phenotype of human airway epithelium by retinoic acid, triiodothyronine, and extracellular matrix. , 1997, American journal of respiratory cell and molecular biology.

[40]  C. Guérin,et al.  Small airway closure and positive end-expiratory pressure in mechanically ventilated patients with chronic obstructive pulmonary disease. , 1997, American journal of respiratory and critical care medicine.

[41]  A. Limper,et al.  Stretch induces cytokine release by alveolar epithelial cells in vitro. , 1999, American journal of physiology. Lung cellular and molecular physiology.

[42]  Karl Grosh,et al.  Microengineered hydromechanical cochlear model. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[43]  W. Seeger,et al.  Surfactant alterations in severe pneumonia, acute respiratory distress syndrome, and cardiogenic lung edema. , 1996, American journal of respiratory and critical care medicine.

[44]  M. Liu,et al.  Stimulation of fetal rat lung cell proliferation in vitro by mechanical stretch. , 1992, The American journal of physiology.

[45]  T. Evans,et al.  Damage to surfactant-specific protein in acute respiratory distress syndrome , 1999, The Lancet.

[46]  M. Saraste,et al.  FEBS Lett , 2000 .

[47]  D. Walters,et al.  Developmental regulation of lung liquid transport. , 2004, Annual review of physiology.

[48]  A. Tanswell,et al.  Mechanical force-induced signal transduction in lung cells. , 1999, American journal of physiology. Lung cellular and molecular physiology.

[49]  J. Grotberg,et al.  Total Liquid Ventilation: Dynamic Airway Pressure and the Development of Expiratory Flow Limitation , 2004, ASAIO journal.

[50]  B. Jonson,et al.  Surfactant dysfunction makes lungs vulnerable to repetitive collapse and reexpansion. , 1997, American journal of respiratory and critical care medicine.

[51]  P. Sterk,et al.  Recurrent exacerbations in severe asthma are associated with enhanced airway closure during stable episodes. , 2000, American journal of respiratory and critical care medicine.

[52]  H. Kuhn,et al.  Apoptosis and necrosis induced by cyclic mechanical stretching in alveolar type II cells. , 2004, American journal of respiratory cell and molecular biology.

[53]  Hideki Fujioka,et al.  The steady propagation of a surfactant-laden liquid plug in a two-dimensional channel , 2005 .

[54]  J B Grotberg,et al.  Respiratory fluid mechanics and transport processes. , 2001, Annual review of biomedical engineering.

[55]  Z. Hantos,et al.  Crackle-sound recording to monitor airway closure and recruitment in ventilated pigs , 2006, European Respiratory Journal.

[56]  Kay C Dee,et al.  Pressure gradient, not exposure duration, determines the extent of epithelial cell damage in a model of pulmonary airway reopening. , 2004, Journal of applied physiology.

[57]  D. Tschumperlin,et al.  Deformation-induced injury of alveolar epithelial cells. Effect of frequency, duration, and amplitude. , 2000, American journal of respiratory and critical care medicine.

[58]  S. Takayama,et al.  Use of Air-Liquid Two-Phase Flow in Hydrophobic Microfluidic Channels for Disposable Flow Cytometers , 2002 .

[59]  J. Schittny,et al.  Spontaneous peristaltic airway contractions propel lung liquid through the bronchial tree of intact and fetal lung explants. , 2000, American journal of respiratory cell and molecular biology.

[60]  S. O'Dea,et al.  Absence of p53 in Clara cells favours multinucleation and loss of cell cycle arrest , 2002, BMC Cell Biology.

[61]  M. Liu,et al.  Differential regulation of extracellular matrix molecules by mechanical strain of fetal lung cells. , 1999, American Journal of Physiology.

[62]  B. Moran,et al.  A system to impose prescribed homogenous strains on cultured cells. , 2001, Journal of applied physiology.