Characterisation of cell adhesion in airway epithelial cell types using electric cell-substrate impedance sensing

Abstract

Research on epithelial cell lines and primary epithelium is required to dissect the mechanisms underlying the structural abnormalities in airway epithelium observed for respiratory diseases, including asthma and chronic obstructive pulmonary disease.The novel electric cell-substrate impedance sensing technique was used to monitor cell adhesion/spreading, barrier function and wound healing. Primary bronchial epithelium was compared with airway epithelial cell lines 16HBE14o-, BEAS-2B, NCI-H292 and A549.BEAS-2B, A549 and primary cells form a confluent monolayer more rapidly than do 16HBE14o-cells. In contrast, 16HBE14o-cells form stronger intercellular contacts, with a 10-fold higher resistance than BEAS-2B, A549 and NCI-H292 cells and a five-fold increase over primary cells. Accordingly, expression of the adhesion molecules zona occludens-1 and E-cadherin was highest in 16HBE14o-cells. These molecules were localised in intercellular junctions in both 16HBE14o-and primary cells. Finally, restoration of barrier function upon injury was impaired in BEAS-2B compared to 16HBE14o-cells.In conclusion, epithelial cell types display remarkable phenotypic differences and should, accordingly, be used to address specific research questions. 16HBE14o-cells appear most suitable for studies on barrier formation, whereas resemblance in attachment of primary and BEAS-2B and A549 cells makes the latter more important for translational research on cell-matrix contact.

Cigarette smoke impairs airway epithelial barrier function and cell-cell contact recovery

“…Using this type of wounding, the epithelial monolayer is able to recover within a few hours, as a result of cell migration/ spreading, and not from proliferation [19]. Barrier function of the 16HBE cells completely recovered within 2 h ( fig.…”

“…Addition of CSE induced a substantial transient fall in low-frequency resistance, the most sensitive parameter to monitor barrier tightness [18,19]. Resistance levels started to decline immediately, with a maximal effect between 4 and 6 h (maximal mean¡SEM decrease 26¡3%; n55), followed by a slow return to control values over ,15 h ( fig.…”

“…We additionally measured the high-frequency capacitance, which is sensitive to changes in cell-matrix contacts, but relatively insensitive to changes in cell-cell contacts [18,19]. Here, CSE exerted less pronounced effects (mean¡SEM maximal increase 9¡1%), indicating that cell-matrix contacts were affected to a smaller extent than cell-cell contacts ( fig.…”

“…Electrical properties of confluent or wounded epithelium were measured using ECIS (Applied Biophysics, Troy, NY, USA) as described previously [18,19]. Upon inoculation, resistance and capacitance were measured at 400 Hz and 40 kHz, respectively.…”

Alterations at the Synthesis and Degradation of E-cadherin in the Human Lungs with Emphysema

“…The inflammation resulting from cigarette smoke exposure, subsequently, the released agents from the inflammatory cells, and the protease-antiprotease imbalance in the lungs lead to the damage of the alveolar epithelium (2,6). The cell death in the alveolar epithelium cells has been shown in the experimental studies where pulmonary emphysema was induced by several substances such as cigarette smoke, elastase, and papain (7,14,16). It has been suggested that alveolar epithelial damage occurs irreversibly and that the repair of alveolar epithelium cannot be done in the pulmonary emphysema (7).…”