Premium
Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propria
Author(s) -
Graupp Matthias,
Gruber HansJürgen,
Weiss Gregor,
Kiesler Karl,
BachnaRotter Sophie,
Friedrich Gerhard,
Gugatschka Markus
Publication year - 2015
Publication title -
the laryngoscope
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.181
H-Index - 148
eISSN - 1531-4995
pISSN - 0023-852X
DOI - 10.1002/lary.25103
Subject(s) - lamina propria , in vitro , macromolecular crowding , medicine , fibrosis , transforming growth factor , pathology , cell , immunology , microbiology and biotechnology , epithelium , macromolecule , biochemistry , chemistry , biology
Objectives/Hypothesis 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. Study Design 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 TGF β −1. Amounts of collagen in the cell layer were significantly higher under crowding conditions with TGF β −1 compared to administration of TGF β −1 alone. Conclusion 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. Level of Evidence NA Laryngoscope , 125:E203–E209, 2015