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

진동이 성대세포주의 세포외기질 변화에 대한 연구

  • Kim, Ji Min (Pusan National University Medical Research Institute, Pusan National University) ;
  • Shin, Sung-Chan (Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Pusan National University and Biomedical Research Institute, Pusan National University Hospital) ;
  • Kwon, Hyun-Keun (Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Pusan National University and Biomedical Research Institute, Pusan National University Hospital) ;
  • Cheon, Yong-Il (Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Pusan National University and Biomedical Research Institute, Pusan National University Hospital) ;
  • Ro, Jung Hoon (Department of Biomedical Engineering, College of Medicine, Pusan National University) ;
  • Lee, Byung-Joo (Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Pusan National University and Biomedical Research Institute, Pusan National University Hospital)
  • 김지민 (부산대학교 의학연구원) ;
  • 신성찬 (부산대학교 의과대학 이비인후과학교실) ;
  • 권현근 (부산대학교 의과대학 이비인후과학교실) ;
  • 천용일 (부산대학교 의과대학 이비인후과학교실) ;
  • 노정훈 (부산대학교 의과대학 의용공학교실) ;
  • 이병주 (부산대학교 의과대학 이비인후과학교실)
  • Received : 2020.11.16
  • Accepted : 2020.12.10
  • Published : 2021.04.30

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.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Science, ICT & Future Planning) (No. 2016R1D1A3B01015539) and clinical research grant from Pusan National University Hospital in 2021.

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