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J Korean Soc Laryngol Phoniatr Logop > Volume 32(1); 2021 > Article
Journal of The Korean Society of Laryngology, Phoniatrics and Logopedics 2021;32(1): 15-23.
doi: https://doi.org/10.22469/jkslp.2021.32.1.15
진동이 성대세포주의 세포외기질 변화에 대한 연구
김지민1 , 신성찬2 , 권현근2 , 천용일2 , 노정훈3 , 이병주2
1부산대학교 의학연구원
2부산대학교 의과대학 이비인후과학교실
3부산대학교 의과대학 의용공학교실
Change of Extracellular Matrix of Human Vocal Fold Fibroblasts by Vibratory Stimulation
Ji Min Kim1 , Sung-Chan Shin2 , Hyun-Keun Kwon2 , Yong-Il Cheon2 , Jung Hoon Ro3 , Byung-Joo Lee2
1Pusan National University Medical Research Institute, Pusan National University, Yangsan, Korea
2Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Pusan National University and Biomedical Research Institute, Pusan National University Hospital, Busan, Korea
3Department of Biomedical Engineering, College of Medicine, Pusan National University, Busan, Korea
Correspondence  Byung-Joo Lee ,Tel: +82-51-240-7528, Fax: +82-51-240-2162, Email: voiceleebj@gmail.com
Received: 16 November 2020; Revised: December 10, 2020   Accepted: 10 December 2020.  Published online: 28 April 2021.
ABSTRACT
Background and Objectives
s 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.
KEY WORDS: Vocal fold fibroblasts; Vibratory bioreactor; Extracellular matrix; Collagen
중심 단어: 생물반응기, 진동자극, 성대세포주, 콜라겐
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