In vitro mechanical vibration down-regulates pro-inflammatory and pro-fibrotic signaling in human vocal fold fibroblasts

PLoS One. 2020 Nov 19;15(11):e0241901. doi: 10.1371/journal.pone.0241901. eCollection 2020.

Abstract

Introduction: Voice rest following phonotrauma or phonosurgery has a considerable clinical impact, but clinical recommendations are inconsistent due to inconclusive data. As biopsies of the vocal folds (VF) for molecular biology studies in humans are unethical, we established a new in vitro model to explore the effects of vibration on human vocal fold fibroblasts (hVFF) in an inflammatory and normal state, which is based on previously published models.

Methods: By using a phonomimetic bioreactor we were able to apply predefined vibrational stress patterns on hVFF cultured under inflammatory or normal conditions. Inflammatory and pro-fibrotic stimuli were induced by interleukin (IL)1β and transforming growth factor (TGF)β1, respectively. Mechanical stimulation was applied four hours daily, over a period of 72 hours. Outcome measurements comprised assessment of extracellular matrix (ECM)-related components, angiogenic factors, and inflammatory and fibrogenic markers on gene expression and protein levels.

Results: Under inflammatory conditions, the inflammatory cytokine IL11, as well as the myofibroblast marker alpha smooth muscle actin (α-SMA) were significantly reduced when additional vibration was applied. The desirable anti-fibrotic ECM component hyaluronic acid was increased following cytokine treatment, but was not diminished following vibration.

Conclusion: Our experiments revealed the effect of vibrational stress on hVFF in an inflammatory state. Elevated levels of certain pro-inflammatory/pro-fibrotic factors could be mitigated by additional vibrational excitation in an in vitro setting. These findings corroborate clinical studies which recommend early voice activation following an acute event.

MeSH terms

  • Actins / metabolism
  • Cells, Cultured
  • Cytokines / pharmacology*
  • Down-Regulation*
  • Extracellular Matrix / drug effects
  • Extracellular Matrix / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects
  • Humans
  • Hyaluronic Acid / metabolism
  • Interleukin-11 / metabolism
  • Interleukin-1beta / pharmacology
  • Models, Biological
  • Signal Transduction / drug effects*
  • Stress, Physiological*
  • Transforming Growth Factor beta1 / pharmacology
  • Vibration
  • Vocal Cords / cytology*
  • Vocal Cords / drug effects
  • Vocal Cords / metabolism

Substances

  • ACTA2 protein, human
  • Actins
  • Cytokines
  • IL11 protein, human
  • Interleukin-11
  • Interleukin-1beta
  • Transforming Growth Factor beta1
  • Hyaluronic Acid

Grants and funding

The authors received no specific funding for this work.