IHC labelling was achieved with the use of the standard Biotin—Streptavidin method to detect: HBD-2 (ab63982, working dilution 1:500, Abcam, Cambridge, UK), HBD-3 (orb183268, working dilution 1:100, Biorbyt Limited, Cambridge, UK), HBD-4 (ab70215, working dilution 1:100, Abcam, Cambridge, UK), IL-10 (orb100193, working dilution 1:600, Biorbyt Limited, Cambridge, UK), and LL-37 (orb88370, working dilution 1:100, Biorbyt LLC, St Louis, MO, USA) [53 (link),54 (link)].
The sample slides were analysed by light microscopy using non-parametric evaluation, which included grading of positively stained tracheal hyaline cartilage cells in the visual field. The results were labelled as follows: 0—no positive cells; 00/+—scant number of positive cells; 0/+—occasional positive cells; +—few positive cells; +/++—few to moderate number of positive cells; ++—moderate number of positive cells; ++/+++—moderate to numerous positive cells; +++—numerous positive cells; +++/++++—numerous to abundant positive cells; ++++—abundant number of positive cells [55 (link)].
The extracellular matrix was also analysed in Bismarck brown and PAS staining for neutral and acidic glycosaminoglycans, and the results were labelled accordingly: -—no positive ground substance; ±—partially stained ground substance; +—stained ground substance.
The sample slides were analysed by light microscopy using non-parametric evaluation, which included grading of positively stained tracheal hyaline cartilage cells in the visual field. The results were labelled as follows: 0—no positive cells; 00/+—scant number of positive cells; 0/+—occasional positive cells; +—few positive cells; +/++—few to moderate number of positive cells; ++—moderate number of positive cells; ++/+++—moderate to numerous positive cells; +++—numerous positive cells; +++/++++—numerous to abundant positive cells; ++++—abundant number of positive cells [55 (link)].
The extracellular matrix was also analysed in Bismarck brown and PAS staining for neutral and acidic glycosaminoglycans, and the results were labelled accordingly: -—no positive ground substance; ±—partially stained ground substance; +—stained ground substance.
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