J Korean Soc Laryngol Phoniatr Logoped.  2021 Apr;32(1):15-23. 10.22469/jkslp.2021.32.1.15.

Change of Extracellular Matrix of Human Vocal Fold Fibroblasts by Vibratory Stimulation

• Kim JM
• Shin SC
• Kwon HK
• Cheon YI
• Ro JH
• Lee BJ

• KMID: 2515638
• DOI: http://doi.org/10.22469/jkslp.2021.32.1.15

Abstract

Background and Objectives
During speech, the vocal folds oscillate at frequencies ranging from 100–200 Hz with amplitudes of a few millimeters. Mechanical stimulation is an essential factor which affects metabolism of human vocal folds. The effect of mechanical vibration on the cellular response in the human vocal fold fibroblasts cells (hVFFs) was evaluated. Materials and Method We created a culture systemic device capable of generating vibratory stimulations at human phonation frequencies. To establish optimal cell culture condition, cellular proliferation and viability assay was examined. Quantitative real time polymerase chain reaction was used to assess extracellular matrix (ECM) related and growth factors expression on response to changes in vibratory frequency and amplitude. Western blot was used to investigate ECM and inflammation-related transcription factor activation and its related cellular signaling transduction pathway.
Results
The cell viability was stable with vibratory stimulation within 24 h. A statistically significant increase of ECM genes (collagen type I alpha 1 and collagen type I alpha 2) and growth factor [transforming growth factor β1 (TGF-β1) and fibroblast growth factor 1 (FGF-1)] observe under the experimental conditions. Vibratory stimulation induced transcriptional activation of NF-κB by phosphorylation of p65 subunit through cellular Mitogen-activated protein kinases activation by extracellular signal regulated kinase and p38 mitogen-activated protein kinases (MAPKs) phosphorylation on hVFFs.
Conclusion
This study confirmed enhancing synthesis of collagen, TGF-β1 and FGF was testified by vibratory stimulation on hVFFs. This mechanism is thought to be due to the activation of NF-κB and MAPKs. Taken together, these results demonstrate that vibratory bioreactor may be a suitable alternative to hVFFs for studying vocal folds cellular response to vibratory vocalization.

Keyword

Vocal fold fibroblasts; Vibratory bioreactor; Extracellular matrix; Collagen; 생물반응기, 진동자극, 성대세포주, 콜라겐