In vivo investigation of acidified pepsin exposure to porcine vocal fold epithelia.

Abstract

The study objective was to investigate epithelial changes in response to direct, repeated, acidified pepsin exposures in an in vivo porcine model. We hypothesized that 12 acidified pepsin applications to simulate reflux would elicit a vocal fold response characterized by inflammation, epithelial proliferation, and increased intercellular space, as well as changes in the gene expression of epithelial junctional proteins, ion transporter proteins, and proinflammatory cytokines. Prospective, in vivo study. Pigs received acidified pepsin (pH = 4) or saline (sham) applied directly to vocal folds. Larynges were collected following three exposures per week for 4 weeks. Vocal fold tissue morphology, collagen, and elastin were evaluated histologically. Gene expression of E-cadherin, zona occludens-1, cystic fibrosis transmembrane conductance regulator, epithelial sodium channel, interleukin-1β, tumor necrosis factor-α, and interferon-γ were measured. Ultrastructural alterations, epithelial intercellular space diameter, and microridge height were measured using transmission electron microscopy. There were no significant differences in histology, gene transcripts, epithelial ultrastructure, intercellular space, and microridge height after acidified pepsin exposure. Twelve simulated reflux challenges were insufficient to elicit epithelial changes, demonstrating the resistance of healthy vocal folds to direct, repeated acidified pepsin exposures. These data increase our understanding of healthy vocal fold defenses and lay the groundwork for a prospective, uninjured, nonsurgical, laryngopharyngeal reflux model where pigs can be exposed directly to acidified pepsin.