Acute In Vitro and In Vivo Effects of Dexamethasone in the Vocal Folds: a Pilot Study

Abstract

Objectives/HypothesisGlucocorticoids (GC)s are commonly employed to treat vocal fold (VF) pathologies. However, VF atrophy has been associated with intracordal GC injections. Dexamethasone‐induced skeletal muscle atrophy is well‐documented in other tissues and believed to be mediated by increased muscle proteolysis via upregulation of Muscle Ring Finger (MuRF)‐1 and Atrogin‐1. Mechanisms of dexamethasone‐mediated VF atrophy have not been described. This pilot study employed in vitro and in vivo models to investigate the effects of dexamethasone on VF epithelium, thyroarytenoid (TA) muscle, and TA‐derived myoblasts. We hypothesized that dexamethasone will increase atrophy‐associated gene expression in TA muscle and myoblasts and decrease TA muscle fiber size and epithelial thickness.Study DesignIn vitro, pre‐clinical.MethodsTA myoblasts were isolated from a female Sprague–Dawley rat and treated with 1 μM dexamethasone for 24‐h. In vivo, 15 New Zealand white rabbits were randomly assigned to three treatment groups: (1) bilateral intracordal injection of 40 μL dexamethasone (10 mg/ml; n = 5), (2) volume‐matched saline (n = 5), and (3) untreated controls (n = 5). Larynges were harvested 7‐days post‐injection. Across in vivo and in vitro experimentation, MuRF‐1 and Atrogin‐1 mRNA expression were measured via RT‐qPCR. TA muscle fiber cross‐sectional area (CSA) and epithelial thickness were also quantified in vivo.ResultsDexamethasone increased MuRF‐1 gene expression in TA myoblasts. Dexamethasone injection, however, did not alter atrophy‐associated gene expression, TA CSA, or epithelial thickness in vivo.ConclusionDexamethasone increased atrogene expression in TA myoblasts, providing foundational insight into GC induced atrophic gene transcription. Repeated dexamethasone injections may be required to elicit atrophy in vivo.Level of EvidenceNA Laryngoscope, 133:2264–2270, 2023

Acute Effects of Systemic Glucocorticoids on the Vocal Folds in a Pre-Clinical Model

“…Thyroarytenoid muscle fiber cross-sectional area (CSA) is a standard technique used to assess skeletal muscle atrophy, 39,40,54,57 and has been described previously by our group. 58 Briefly, a ROI outlining muscle fibers in H&E stained samples was acquired. "Default" thresholding was performed and the particle analysis tool was utilized to calculate total muscle fiber area in each sample.…”

A Novel Method for Thyroarytenoid Myofiber Culture

“…In vitro models involving animal-derived vocal fold (VF) myoblasts or myotubes (i.e., precursors to mature myofibers) have been employed to increase mechanistic insight regarding VF muscle health and physiology. [3][4][5] Although in vitro investigation offers affordability and precise experimental control, elucidation of the innate cellular, biochemical, and biomechanical interactions in organ systems is lacking, limiting translation to clinical scenarios. The development and optimization of methods to investigate intrinsic laryngeal musculature structure and function have the potential to alter and enhance clinical paradigms.…”

“…The development of novel therapeutic strategies is a public health priority with the potential to profoundly impact millions of patients with laryngeal disease and voice‐related disabilities. In vitro models involving animal‐derived vocal fold (VF) myoblasts or myotubes (i.e., precursors to mature myofibers) have been employed to increase mechanistic insight regarding VF muscle health and physiology 3–5 . Although in vitro investigation offers affordability and precise experimental control, elucidation of the innate cellular, biochemical, and biomechanical interactions in organ systems is lacking, limiting translation to clinical scenarios.…”