Acute Nanoparticle Exposure to Vocal Folds: A Laboratory Study

Abstract

Airway exposure to nanoparticles is common in occupational settings. Inhaled nanoparticles have toxic effects on respiratory tissue. Vocal folds are also at direct risk from inhaled nanoparticles. This study investigated the effects of single-walled carbon nanotubes (SWCNT), a type of nanoparticle, on vocal fold epithelium and fibroblasts. These cell types were selected for study as the epithelium is the outer layer of the vocal folds and fibroblasts are the most common cell type in connective tissue underlying the epithelium. Native porcine vocal fold epithelium and cultured human vocal fold fibroblasts were exposed to SWCNTs (100 ng/mL) and control (no SWCNT) in vitro. Epithelial and fibroblast viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Epithelial barrier integrity was assessed with transepithelial resistance and sodium fluorescein permeability. Epithelial tight junctional protein occludin expression was measured with Western blot. Gene expressions of the fibroblast-specific protein 1 (FSP-1), α-smooth muscle actin (α-SMA), and collagen III (Col-III) were assessed using quantitative polymerase chain reaction. Transcriptional expression of genes encoding FSP-1 and Col-III was increased significantly following SWCNT exposure. There were no significant differences between control and SWCNT groups on any of the other measures. SWCNT exposure induces vocal fold fibroblasts to a fibrotic phenotype. These data help us understand vocal fold defense mechanisms and lay the groundwork for studying the physiological effects of nanoparticle exposure in vivo.