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Chronic mucosal and submucosal injury can lead to persis- tent inflammation and tissue remodeling. We hypothesized that mi- crostructural and mechanical properties of the airway wall could be derived from multiphoton images. New Zealand White rabbits were intubated, and the tracheal epithelium gently denuded every other day for five days three injuries. Three days following the last injury, the tracheas were excised for multiphoton imaging, mechanical com- pression testing, and histological analysis. Multiphoton imaging and histology confirm epithelial denudation, mucosal ulceration, subepi- thelial thickening, collagen deposition, immune cell infiltration, and a disrupted elastin network. Elastase removes the elastin network and relaxes the collagen network. Purified collagenase removes epithe- lium with subtle subepithelial changes. Young's modulus E mea- sured in kiloPascal was significantly elevated for the scrape injured 9.0±3.2 trachea, and both collagenase 2.6±0.4 and elastase 0.8±0.3 treatment significantly reduced E relative to control 4.1±0.7. E correlates strongly with second harmonic generation SHG signal depth decay for enzyme-treated and control tracheas R 2 =0.77, but not with scrape-injured tracheas. We conclude that E of subepithelial connective tissue increases on repeated epithelial wounding, due in part to changes in elastin and collagen microstruc- ture and concentration. SHG depth decay is sensitive to changes in extracellular matrix content and correlates with bulk Young's modulus. © 2010 Society of Photo-Optical Instrumentation Engineers.

Linking optics and mechanics in an in vivo model of airway fibrosis and epithelial injury