Structural changes in the extracellular matrix after cross-linking of nasal polyp tissue

Chronic rhinosinusitis with nasal polyposis is a disease characterised by a mechanical dysfunction that facilitates anomalous growth of the nasal mucosa, due to its unique remodelling process. The development of procedures capable of interfering with this process is thus of the utmost importance. This study deals with the evaluation of the effects of cross-linking upon the nasal polyp tissue extracellular matrix. Six patients undergoing surgery for polypectomy were selected. The riboflavin/UVA collagen cross-linking technique was applied to the surface of epithelialised and de-epithelialised resected polyps of the intervention group. The control group polyps were not submitted to cross-linking procedures. Ultrathin polyp tissue sections (8 μm) were processed for immunofluorescence with mouse anti-type I collagen antibody and AlexaFluor 488 conjugated secondary antibody plus DAPI counterstaining, and analysed by confocal microscopy. The effect of riboflavin/UVA collagen cross-linking was visible on confocal fluorescence microscopy. Quantitative morphology was associated with fluorescence imaging analysis, and pixel density and brightness were evaluated. The surface of treated polyps exhibited a higher density of collagen fibres compared to control polyps, as could be observed both visually and through objective measurements of the fluorescent regions. The effect was enhanced on the surface of the de-epithelialised polyps. The higher density of collagen fibres exhibited by the de-epithelialised treated polyps demonstrates the feasibility of this technique in interfering with the remodelling process and the mechanical dysfunction found in chronic rhinosinusitis with nasal polyposis.