title: Utility of cell viability assays for use with ex vivo vocal fold epithelial tissue.
abstract: OBJECTIVE
  Ex vivo models are routinely used to investigate the barrier function of the vocal fold epithelium. However, there are limited reports on assays that can be used to investigate the effect of clinically relevant challenges on vocal fold epithelial tissue viability. Our objective was to determine the utility of two assays routinely used in cell culture-a cellular metabolic activity assay and a cell membrane integrity assay-to investigate the viability of ex vivo porcine vocal fold epithelium.
  
  
  METHODS
  Prospective, ex vivo animal study.
  
  
  METHODS
  Porcine vocal folds were exposed to acrolein, hydrochloric acid, or hydrogen peroxide challenge. An untreated, sham challenge was included as a control. Assays including metabolic activity, cell membrane integrity, and histology were used to determine whether challenges reduced epithelial viability as compared to sham.
  
  
  RESULTS
  Cell membrane integrity and metabolic activity assays detected reductions in viability following hydrochloric acid and hydrogen peroxide challenges but not acrolein challenge as compared to sham. No challenge produced significant changes in epithelial appearance as evidenced by light microscopy.
  
  
  CONCLUSIONS
  Metabolic activity and cell membrane integrity assays are valuable tools that can be used to evaluate the viability of ex vivo vocal fold epithelial tissue following clinically relevant challenges. As viability is reduced, the ability of epithelial tissue to maintain its barrier function is compromised. Accurate assessment of viability may provide us clues into understanding mechanisms underlying vocal fold epithelial injury and disease.
  
  
  METHODS
  NA Laryngoscope, 125:E180-E185, 2015.
authors.affiliation: Department of Surgery, Division of Otolaryngology-Head & Neck Surgery, University of Wisconsin-Madison, Madison, Wisconsin.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Erickson-DiRenzo
authors.forename: M Preeti
authors.initials: MP
authors.lastname: Sivasankar
authors.forename: Susan L
authors.initials: SL
authors.lastname: Thibeault
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism; D005260:Female; D007820:Laryngeal Mucosa / Q000166:cytology* / Q000378:metabolism; D008297:Male; D011446:Prospective Studies; D013552:Swine; D014827:Vocal Cords / Q000166:cytology* / Q000378:metabolism
keywords: Ex vivo model; viability assay; vocal fold epithelium