The effects of gas humidification with high-flow nasal cannula on cultured human airway epithelial cells

Aaron Chidekel, Yan Zhu, Jordan Wang, John J Mosko, Elena Rodriguez, Thomas H Shaffer, Aaron Chidekel, Yan Zhu, Jordan Wang, John J Mosko, Elena Rodriguez, Thomas H Shaffer

Abstract

Humidification of inspired gas is important for patients receiving respiratory support. High-flow nasal cannula (HFNC) effectively provides temperature and humidity-controlled gas to the airway. We hypothesized that various levels of gas humidification would have differential effects on airway epithelial monolayers. Calu-3 monolayers were placed in environmental chambers at 37°C with relative humidity (RH) < 20% (dry), 69% (noninterventional comparator), and >90% (HFNC) for 4 and 8 hours with 10 L/min of room air. At 4 and 8 hours, cell viability and transepithelial resistance measurements were performed, apical surface fluid was collected and assayed for indices of cell inflammation and function, and cells were harvested for histology (n = 6/condition). Transepithelial resistance and cell viability decreased over time (P < 0.001) between HFNC and dry groups (P < 0.001). Total protein secretion increased at 8 hours in the dry group (P < 0.001). Secretion of interleukin (IL)-6 and IL-8 in the dry group was greater than the other groups at 8 hours (P < 0.001). Histological analysis showed increasing injury over time for the dry group. These data demonstrate that exposure to low humidity results in reduced epithelial cell function and increased inflammation.

Figures

Figure 1
Figure 1
Cell-culture environmental chambers. Transwell plates were exposed to one of three levels of relative humidity: 90% (HFNC) for 4 and 8 hours with 10 L/pm of room air at 37°C.
Figure 2
Figure 2
Transepithelial resistance (TER) for Calu-3 monolayers exposed to one of three levels of relative humidity: 90% (HFNC). TER was less than the noninterventional comparator group in the HFNC and dry groups (P < 0.001). Data are mean ± SEM. *Group effect (P < 0.001). §Time effect (P < 0.001).
Figure 3
Figure 3
Cell viability for Calu-3 monolayers exposed to one of three levels of relative humidity: 90% (HFNC). CellTiter Blue cell viability assay was performed to estimate the number of viable cells and determine cell viability by the intensity of fluorescence. Cell viability decreased over time (P < 0.001) for HFNC and dry groups. Data are mean ± SEM. *Group effect (P < 0.05). §Time effect (P < 0.05). n = 6/condition.
Figure 4
Figure 4
Total protein concentration in apical surface wash fluid from Calu-3 monolayers exposed to one of three levels of relative humidity: 90% (HFNC). Secretion of total protein increased at 8 hours in the dry group (P < 0.001), but there was no difference between the noninterventional comparator group and the HFNC groups (P > 0.05) and no difference between groups at 4 hours (P > 0.05). Data are mean ± SEM. *Group effect (P < 0.05). §Time effect (P < 0.05). n = 6/condition.
Figure 5
Figure 5
Proinflammatory mediator concentration in apical surface wash fluid from Calu-3 monolayers exposed to one of three levels of relative humidity: 90% (HFNC). Secretion of interleukin (IL)-6 (5A) and IL-8 (5B) in the dry group was greater than the noninterventional comparator group and the HFNC groups at 8 hours (P < 0.001), with no difference between the noninterventional comparator group and the HFNC groups (P > 0.05) and no difference between any groups at 4 hours (P > 0.05). Data are mean ± SEM. *Group effect (P < 0.05). §Time effect (P < 0.05).
Figure 6
Figure 6
Cytomorphological examination of Calu-3 cell monolayers exposed to one of three levels of relative humidity: 90% (HFNC). Representative cytomorphological examination of Calu-3 cells for both the noninterventional comparator group and the HFNC group demonstrated normal morphology. At 4 and 8 hours, the dry group showed abnormal cellular appearances, swollen nuclei, intracellular and nuclear vacuoles, diffused cytoplasm, and cellular debris. All cytospins were examined by light microscopy at 40x magnification.
Figure 7
Figure 7
Semi-quantitative assessment of cell morphology as the ratio of abnormal cells to total cells. Histological analysis did not show injury for the noninterventional comparator or the high flow nasal cannula groups at 4 and 8 hours, whereas the dry group demonstrated increasing injury over time. Data are mean ± SEM. *Group effect (P < 0.05). §Time effect (P < 0.05). n = 6/condition.

References

    1. Shelly MP, Lloyd GM, Park GR. A review of the mechanisms and methods of humidification of inspired gases. Intensive Care Medicine. 1988;14(1):1–9.
    1. Fulmer JD, Snider GL, Albert RK. ACCP-NHLBI national conference on oxygen therapy. Chest. 1984;86(2):234–247.
    1. Campbell EJ, Baker MD, Crites-Silver P. Subjective effects of humidification of oxygen for delivery by nasal cannula: a prospective study. Chest. 1988;93(2):289–293.
    1. Shelly MP. The humidification and filtration functions of the airways. Respiratory Care Clinics of North America. 2006;12(2):139–148.
    1. McFadden ER., Jr. Heat and water exchange in human airways. American Review of Respiratory Disease. 1992;146:S8–S10.
    1. McFadden ER, Jr., Pichurko BM, Bowman HF. Thermal mapping of the airways in humans. Journal of Applied Physiology. 1985;58(2):564–570.
    1. Kubicka ZJ, Limauro J, Darnall RA. Heated, humidified high-flow nasal cannula therapy: yet another way to deliver continuous positive airway pressure? Pediatrics. 2008;121(1):82–88.
    1. Greenspan JS, Wolfson MR, Shaffer TH. Airway responsiveness to low inspired gas temperature in preterm neonates. Journal of Pediatrics. 1991;118(3):443–445.
    1. On LS, Boonyongsunchai P, Webb S, Davies L, Calverley PMA, Costello RW. Function of pulmonary neuronal M2 muscarinic receptors in stable chronic obstructive pulmonary disease. American Journal of Respiratory and Critical Care Medicine. 2001;163(6):1320–1325.
    1. Doyle A, Joshi M, Frank P, Craven T, Moondi P, Young P. A change in humidification system can eliminate endotracheal tube occlusion. Journal of Critical Care. 2011;26:673.e1–673.e4.
    1. Branson RD, Gentile MA. Is humidification always necessary during noninvasive ventilation in the hospital? Respiratory Care. 2010;55(2):209–216.
    1. Shen BQ, Finkbeiner WE, Wine JJ, Mrsny RJ, Widdicombe JH. Calu-3: a human airway epithelial cell line that shows cAMP-dependent Cl- secretion. American Journal of Physiology. 1994;266(5):L493–L501.
    1. Da Paula AC, Ramalho AS, Farinha CM, et al. Characterization of novel airway submucosal gland cell models for cystic fibrosis studies. Cellular Physiology and Biochemistry. 2005;15(6):251–262.
    1. Joo NS, Lee DJ, Winges KM, Rustagi A, Wine JJ. Regulation of antiprotease and antimicrobial protein secretion by airway submucosal gland serous cells. Journal of Biological Chemistry. 2004;279(37):38854–38860.
    1. Zhang Y, Reenstra WW, Chidekel A. Antibacterial activity of apical surface fluid from the human airway cell line Calu-3: pharmacologic alteration by corticosteroids and β2-agonists. American Journal of Respiratory Cell and Molecular Biology. 2001;25(2):196–202.
    1. Grainger CI, Greenwell LL, Lockley DJ, Martin GP, Forbes B. Culture of Calu-3 cells at the air interface provides a representative model of the airway epithelial barrier. Pharmaceutical Research. 2006;23(7):1482–1490.
    1. Zhu Y, Chidekel A, Shaffer TH. Cultured human airway epithelial cells (Calu-3): a model of human respiratory function, structure, and inflammatory responses. Critical Care Research and Practice. 2010;2010:8 pages.394578
    1. Forbes B, Ehrhardt C. Human respiratory epithelial cell culture for drug delivery applications. European Journal of Pharmaceutics and Biopharmaceutics. 2005;60(2):193–205.
    1. Sporty JL, Horálková L, Ehrhardt C. In vitro cell culture models for the assessment of pulmonary drug disposition. Expert Opinion on Drug Metabolism and Toxicology. 2008;4(4):333–345.
    1. Hirakata Y, Yano H, Arai K, et al. Monolayer culture systems with respiratory epithelial cells for evaluation of bacterial invasiveness. Tohoku Journal of Experimental Medicine. 2010;220(1):15–19.
    1. Devor DC, Singh AK, Lambert LC, DeLuca A, Frizzell RA, Bridges RJ. Bicarbonate and chloride secretion in Calu-3 human airway epithelial cells. Journal of General Physiology. 1999;113(5):743–760.
    1. Winton HL, Wan H, Cannell MB, et al. Cell lines of pulmonary and non-pulmonary origin as tools to study the effects of house dust mite proteinases on the regulation of epithelial permeability. Clinical and Experimental Allergy. 1998;28(10):1273–1285.
    1. Stewart CE, Torr EE, Mohd Jamili NH, Bosquillon C, Sayers I. Evaluation of differentiated human bronchial epithelial cell culture systems for asthma research. Journal of Allergy. 2012;2012:11 pages.943982
    1. Babu PBR, Chidekel A, Shaffer TH. Hyperoxia-induced changes in human airway epithelial cells: the protective effect of perflubron. Pediatric Critical Care Medicine. 2005;6(2):188–194.
    1. Zhu Y, Miller TL, Singhaus CJ, Shaffer TH, Chidekel A. Effects of oxygen concentration and exposure time on cultured human airway epithelial cells. Pediatric Critical Care Medicine. 2008;9(2):224–229.
    1. Foster KA, Avery ML, Yazdanian M, Audus KL. Characterization of the Calu-3 cell line as a tool to screen pulmonary drug delivery. International Journal of Pharmaceutics. 2000;208(1-2):1–11.
    1. Mathias NR, Timoszyk J, Stetsko PI, Megill JR, Smith RL, Wall DA. Permeability characteristics of Calu-3 human bronchial epithelial cells: in vitro-in vitro correlation to predict lung absorption in rats. Journal of Drug Targeting. 2002;10(1):31–40.
    1. Chang GY, Cox CA, Shaffer TH. Nasal cannula, CPAP, and high-flow nasal cannula: effect of flow on temperature, humidity, pressure, and resistance. Biomedical Instrumentation and Technology. 2011;45(1):69–74.
    1. Coalson JJ. Pathology of new bronchopulmonary dysplasia. Seminars in Neonatology. 2003;8(1):73–81.
    1. Munshi UK, Niu JO, Siddiq MM, Parton LA. Elevation of interleukin-8 and interleukin-6 precedes the influx of neutrophils in tracheal aspirates from preterm infants who develop bronchopulmonary dysplasia. Pediatric Pulmonology. 1997;24:331–336.
    1. Tullus K, Noack GW, Burman LG, Nilsson R, Wretlind B, Brauner A. Elevated cytokine levels in tracheobronchial aspirate fluids from ventilator treated neonates with bronchopulmonary dysplasia. European Journal of Pediatrics. 1996;155(2):112–116.
    1. Krause MF, Wiemann T, Reisner A, Orlowska-Volk M, Köhler H, Ankermann T. Surfactant reduces extravascular lung water and invasion of polymorphonuclear leukocytes into the lung in a piglet model of airway lavage. Pulmonary Pharmacology and Therapeutics. 2005;18(2):129–139.
    1. Baier RJ, Loggins J, Kruger TE. Monocyte chemoattractant protein-1 and interleukin-8 are increased in bronchopulmonary dysplasia: relation to isolation of ureaplasma urealyticum. Journal of Investigative Medicine. 2001;49(4):362–369.
    1. Baier RJ, Loggins J, Kruger TE. Increased interleukin-8 and monocyte chemoattractant protein-1 concentrations in mechanically ventilated preterm infants with pulmonary hemorrhage. Pediatric Pulmonology. 2002;34(2):131–137.
    1. Oshodi A, Dysart K, Cook A, et al. Airway injury resulting from repeated endotracheal intubation: possible prevention strategies. Pediatric Critical Care Medicine. 2011;12(1):e34–e39.
    1. Chong E, Dysart KC, Chidekel A, Locke R, Shaffer TH, Miller TL. Heat shock protein 70 secretion by neonatal tracheal tissue during mechanical ventilation: association with indices of tissue function and modeling. Pediatric Research. 2009;65(4):387–391.

Source: PubMed

Sponsors and Collaborators

Medical Conditions

Drug Interventions

3
Subscribe