Fig. 1
a Binary images of day 28 mucociliary BECs treated with control vehicle (CoV), mesoporous graphitized nanocarbon (NG) or multi-walled carbon nanotubes (MWCNTs) for 24-h. Cells were treated while undifferentiated, and allowed to differentiate in air-liquid interface for 28 days after ending treatment. A “post-ALI” MWCNT 1μg/cm2 treatment is included for comparison, in which treatment was delayed until after the BECs had differentiated for 20 days in ALI. Tubulin staining in cilia (top) and F-actin staining in tight junctions (bottom) demonstrate reduction of differentiated cells resulting from MWCNT treatment. These images are examples of those used for pixel density analysis, representing a summation image from the top 10 μm of epithelium. Intensity below a threshold value was excluded from the binary image. b Results of image quantification of randomized sections of membrane. There is a significant (*p < 0.05, **p < 0.01 ANOVA, Bonferroni post hoc) reduction in α-tubulin and F-actin translocation to cilia and tight junctions (respectively) following MWCNT treatment. Data compiled from 3 donors, with 2–3 replicate wells per donor. Bars represent mean ± standard deviation (SD)