Hydrogel-Supported, Engineered Model of Vocal Fold Epithelium

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Cell Biology

Background:

• A functional engineered model of vocal fold epithelium is crucial for understanding vocal fold health and disease.
• Current models lack the complexity to fully replicate native vocal fold tissue characteristics.

Purpose of the Study:

• To develop an engineered vocal fold epithelium model using primary porcine vocal fold epithelial cells (VFECs).
• To investigate the roles of matrix stiffness, composition, and paracrine signaling in VFEC behavior and stratification.

Main Methods:

• Primary VFECs were isolated from porcine larynxes and cultured on hyaluronic acid-derived synthetic basement membranes.
• Matrices were functionalized with integrin-binding (RGDSP) and syndecan-binding (AG73) peptides.
• Matrix stiffness was modulated, and cytokine supplementation from fibroblasts was employed.
• Engineered tissue was analyzed for morphology, cell-cell junctions, protein expression, and basement membrane secretion.

Main Results:

• Matrix stiffness and peptide composition cooperatively regulated VFEC adhesion, proliferation, and stratification.
• Physiological stiffness promoted cobblestone morphology, while softer matrices induced spindle-shaped cells.
• Both RGDSP and AG73 peptides were required for stratified epithelium development.
• Cytokine supplementation led to 4-5 distinct cell layers, resembling native tissue.

Conclusions:

• Engineered vocal fold epithelium closely mimics native tissue morphology and molecular markers.
• Stiffness, cell-matrix interactions, and paracrine signaling are key factors in VFEC stratification.
• This engineered model serves as a versatile tool for vocal fold research.