title: Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propria.
abstract: OBJECTIVE
  Vocal fold fibrosis represents a major disease burden. Screening of antifibrotic compounds could be facilitated by an in vitro fibrogenesis system. Limitations of existing models might be overcome by implication of the excluded volume effect.
  
  
  METHODS
  In-vitro study.
  
  
  METHODS
  Vocal fold fibroblasts obtained from rats' lamina propria were cultured in four different settings: in standard medium, under "crowded" conditions by adding inert macromolecules, under external administration of transforming growth factor (TGF)ß-1, and under a combination of both. After 5 days, supernatant and cell layer were collected and analyzed by enzyme-linked immunosorbent assay. Immunofluorescence was additionally performed.
  
  
  RESULTS
  Collagen-alpha1(I) deposition increased significantly under crowded conditions and after administration of TGFβ-1. Amounts of collagen in the cell layer were significantly higher under crowding conditions with TGFβ-1 compared to administration of TGFβ-1 alone.
  
  
  CONCLUSIONS
  Crowding enhanced collagen deposition, resulting in more favorable conditions for studying fibrogenesis. This can be the first step toward developing a robust in vitro model for testing antifibrotic compounds.
  
  
  METHODS
  NA.
authors.affiliation: ENT University Hospital Graz, Department of Phoniatrics, Medical University Graz, Graz, Austria.
authors.forename: Matthias
authors.initials: M
authors.lastname: Graupp
authors.affiliation: Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University Graz, Graz, Austria.
authors.forename: Hans-Jürgen
authors.initials: HJ
authors.lastname: Gruber
authors.affiliation: Institute of Cell Biology, Histology and Embryology, Medical University Graz, Graz, Austria.
authors.forename: Gregor
authors.initials: G
authors.lastname: Weiss
authors.affiliation: ENT University Hospital Graz, Department of Phoniatrics, Medical University Graz, Graz, Austria.
authors.forename: Karl
authors.initials: K
authors.lastname: Kiesler
authors.affiliation: ENT University Hospital Graz, Department of Phoniatrics, Medical University Graz, Graz, Austria.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bachna-Rotter
authors.affiliation: ENT University Hospital Graz, Department of Phoniatrics, Medical University Graz, Graz, Austria.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Friedrich
authors.affiliation: ENT University Hospital Graz, Department of Phoniatrics, Medical University Graz, Graz, Austria.
authors.forename: Markus
authors.initials: M
authors.lastname: Gugatschka
mesh_terms: D000818:Animals; D035843:Biomedical Research; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts; D005355:Fibrosis; D046911:Macromolecular Substances; D008297:Male; D008954:Models, Biological; D009092:Mucous Membrane / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014827:Vocal Cords / Q000473:pathology*
chemical_list: D046911:Macromolecular Substances; D003094:Collagen
keywords: Vocal fold scar; fibrosis; in vitro fibrogenesis; macromolecular crowding