Concentration Effects of Methylprednisolone in Human Vocal Fold Fibroblast–Macrophage Co‐Culture

• Published in: The Laryngoscope 2023-11, Vol.133 (11), p.3116-3122
• Main Authors: Nakamura, Ryosuke, Bing, Renjie, Gartling, Gary J., Garabedian, Michael J., Branski, Ryan C.
• Format: Article
• Language: English
• Subjects: Cells, Cultured; Coculture Techniques; Cyclooxygenase 2 - metabolism; Fibroblasts; Fibroblasts - metabolism; Fibrosis; glucocorticoids; Glucocorticoids - pharmacology; Humans; inflammation; larynx; Lipopolysaccharides; macrophages; Macrophages - metabolism; Methylprednisolone - pharmacology; steroids; Vocal Cords - pathology; vocal fold; voice
• ISSN: 0023-852X; 1531-4995; 1531-4995
• DOI: 10.1002/lary.30763

Objective The diversity of glucocorticoid (GC) properties may underlie variability of clinical efficacy for vocal fold (VF) disease. Optimized therapeutic approaches must account for tissue complexity as well as interactions between cell types. We previously reported that reduced GC concentrations inhibited inflammation without eliciting fibrosis in mono‐cultured VF fibroblasts and macrophages. These data suggested that a refined approach to GC concentration may improve outcomes. In the current study, co‐culture of VF fibroblasts and macrophages was employed to investigate the effects of different concentrations of methylprednisolone on fibrotic and inflammatory response genes in VF fibroblasts to optimize management paradigms. Study Design In vitro. Methods THP‐1 monocyte‐derived macrophages were stimulated with interferon‐γ (IFN‐γ), lipopolysaccharide (LPS), or transforming growth factor‐β (TGF‐β) to induce inflammatory (M(IFN/LPS)) and fibrotic (M(TGF)) phenotypes. Macrophages were then co‐cultured with a human VF fibroblast cell line using a 0.4 μm pore membrane with or without 0.1–3000 nM methylprednisolone. Inflammatory (CXCL10, TNF, and PTGS2) and fibrotic (ACTA2, CCN2, and COL1A1) gene expression was quantified in fibroblasts. Results Incubating VF fibroblasts with M(IFN/LPS) macrophages increased expression of TNF and PTGS2, and this effect was inhibited by methylprednisolone. Incubation of VF fibroblasts with M(TGF) macrophages increased expression of ACTA2, CCN2, and COL1A1, and this effect was enhanced by methylprednisolone. The concentration of methylprednisolone required to downregulate inflammatory genes (TNF and PTGS2) was lower than that to upregulate fibrotic genes (ACTA2, CCN2, and COL1A1). Conclusion Reduced concentration of methylprednisolone effectively suppressed inflammatory genes without enhancing fibrotic genes, suggesting that a refined approach to GC concentration may improve clinical outcomes. Level of Evidence N/A Laryngoscope, 133:3116–3122, 2023 This work provides additional in vitro support for optimizing glucocorticoid dosing when treating patients with vocal fold pathology. Increased concentrations enhanced the fibrotic response in vocal fold fibroblasts induced by polarized macrophages.