Table 1.
Comparison between 3D in vitro model of reconstituted epithelium and organotypic explant culture.
Figure 1.
Ultrastructural study of apical surfaces in ALI cultures from both nasal polyp and control nasal mucosa at different days by scanning electron microscopy.
At day 0 (left column), representative images are showing apical surfaces without ciliated cells. At days 14 (centre column) and 28 (right column), apical surfaces showing ciliated cells.
Figure 2.
Ultrastructural study in ALI cultures from both nasal polyp and control nasal mucosa at different days by transmission electron microscopy.
At day 0 (left column), representative images are showing desmosomes (arrowheads), interdigitations (asteriks). At day 14 (centre column), ciliated (C), goblet (G) and basal (B) cells. At day 28 (right column), longitudinal sections of motile cilia (upper) and secretory granules of a goblet cell (lower).
Figure 3.
Re-differentation over time of human nasal epithelial cells from both nasal polyp and control nasal mucosa in ALI cultures.
A, Pie charts, with modeled size relative to total cell number, are representing the percentage of different cell phenotypes over time. B, table shows the total number of cells and percentages of cell phenotypes from both cultures types. Total and positive cells were counted in three wells (5 fields from different areas of each well at 400x magnification) from different nasal polyp (NP, n = 3) and control nasal mucosa (NM, n = 3) ALI cultures and the results were subsequently averaged. *, P<0.05 vs day 0. The nonparametric Wilcoxon Signed-Ranks Test and Mann-Whitney U test were used for paired comparisons and between-group comparisons respectively. Statistical significance was set at P<0.05.
Figure 4.
Representative immunocytochemical images of different cell phenotypes in both undifferenciated (day 0) and fully differentiated (day 28) cultures from both nasal polyp and control nasal mucosa.
A, Basal progenitor/stem cells, with positive immunoreactivity to ΔNp63. B, Goblet cells, with positive immunoreactivity to MUC5AC. C, Ciliated cells, with positive immunoreactivity to β-tubulin IV. D, Ciliated cells, marked by β-tubulin IV+ (green), and goblet cells, marked by MUC5B+ (red, arrowheads). Final magnification x400.
Figure 5.
Overtime secretion of mucin and lactoferrin in nasal polyp (black boxes) and control nasal mucosa (white boxes) ALI cultures.
A, MUC5AC secretion (ng/ml). B, MUC5B secretion (RU/ml). C, Lactoferrin secretion (ng/ml). Results are expressed as medians and interquartile ranges with 25th–75th percentiles. *, P<0.05 vs day 0. Statistical analysis by Mann-Whitney U test (between groups) and by Wilcoxon test (within groups).
Figure 6.
Cytokine and chemokine apical secretion from both nasal polyp (black boxes) and control nasal mucosa (white boxes) ALI cultures at different days.
A, interleukin-8 (IL-8), B, granulocyte-macrophage colony-stimulating factor (GM-CSF), C, eotaxin 1, D, monocyte chemoattractant protein-1 (MCP-1), E, interleukin-6 (IL-6), and F, interferon-γ–induced protein-10 (IP-10), levels in apical washes. Results are expressed as medians and interquartile ranges with 25th and 75th percentiles. ns, P>0.05 vs day 0 (100%); *, P<0.05 vs day 0. Statistical analysis by Mann-Whitney U test (between groups) and by Wilcoxon test (within groups).