액상형 비열대기 플라즈마(Non-thermal atmospheric plasma treated solution)의 상기도 점막 조직 재생에 대한 효과 확인 및 평가
Identification and evaluation of regenerative effect on the upper airway mucosa of non-thermal atmospheric plasma treated solution
- 주제(키워드) Non-thermal atmospheric plasma treated solution (NTS) , Regeneration , Nasal mucosa , Vocal fold mucosa , Wound healing
- 발행기관 아주대학교
- 지도교수 김철호
- 발행년도 2020
- 학위수여년월 2020. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000029700
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Plasma is a mixture of electrons, ions, and energetic photons in the form of ionized gas that is generated when the gas is further energized. Non-thermal atmospheric plasma has several medical effects such as sterilization, anti-cancer, anti-inflammation, and tissue regeneration. However, the existing probe type is limited to local direct treatment. Therefore, we investigated the various effects using liquid type plasma to overcome this limitation. In addition, the therapeutic effects of non-thermal plasma treated solution (NTS) on upper airway mucosa have yet to be determined. In the first part, the effect of NTS on the regeneration of nasal mucosa was studied. Experiments were carried out using BEAS-2B, a human bronchial epithelial cell line similar to nasal mucosa epithelium. NTS had no cytotoxicity to the BEAS-2B cells and enhanced cell proliferation. NTS also promoted migration of BEAS-2B cells. NTS increased cell proliferation and migration via epidermal growth factor receptor activities and epithelial-to-mesenchymal transition signaling. Furthermore, NTS enhanced wound healing of nasal mucosa in an animal model. In the second part, the effect of NTS on the regeneration of vocal fold mucosa was examined. Migration and matrix metalloproteinase-2 expression of lipopolysaccharide (LPS)-treated human vocal fold-derived mesenchymal stem cells (VF-MSCs) were enhanced by NTS treatment. NTS treatment not only ameliorated nuclear factor-κB and interleukin-6 activation, induced by NTS treatment, but also the increased manifestation of α-smooth muscle actin and fibronectin, induced by transforming growth factor-ß. In a rabbit vocal fold scarring animal model, histological analyses showed increased hyaluronic acid deposition and decreased collagen accumulation after NTS injection. Videokymographic analysis showed more improved vibrations in NTS-treated vocal fold mucosa compared to those in non-treated group. The results of this study suggest that NTS may improve regeneration of the upper airway mucosa and improve functional remodeling following damage to the upper airway mucosa.
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Table of Contents
Abstract ⅰ
Table of Contents ⅲ
List of Figures ⅵ
Part I
Ⅰ. Introduction 2
Ⅱ. Materials and Methods 3
A. Cell culture 3
B. Design of non-thermal plasma treated solution (NTS) 3
C. Cell cytotoxicity activity (MTT assay) 4
D. Cell proliferation activity (BrdU assay) 5
E. Scratch wound healing assay 5
F. Transwell migration assay 5
G. Western blot 6
H. Immunofluorescence assay 6
I. Quantitative real-time PCR 7
J. Gelatin zymogram assay 7
K. In vivo study 8
L. Immuno-histochemical analysis 9
M. Statistical analysis 9
Ⅲ. Results 11
A. NTS has no cytotoxicity to the BEAS-2B cells and enhances cell proliferation 11
B. NTS enhances migration of BEAS-2B cells 12
C. NTS increases cell proliferation and migration via epidermal growth factor receptor (EGFR) activities 14
D. NTS increases cell migration via the epithelial-to-mesenchymal transition (EMT) signaling 15
E. NTS enhances matrix metalloproteinases-2 (MMP-2)/matrix metalloproteinases-9 (MMP-9) activities 18
F. NTS enhances nasal mucosa wound healing in an animal model 20
Ⅳ. Discussion 23
Ⅴ. Conclusion 27
Ⅵ. References 28
Part II
Ⅰ. Introduction 34
Ⅱ. Materials and Methods 36
A. Cell culture 36
B. Fluoresce-activated cell sorting analysis 36
C. Design of non-thermal plasma treated solution (NTS) 36
D. Cell viability assay (MTT assay) 37
E. Scratch wound healing assay 37
F. Quantitative real-time PCR 37
G. Western blotting 38
H. Reverse transcription PCR 38
I. Animal model and surgical techniques 39
J. Endoscopic and histologic analysis 39
K. Immunohistochemical analysis 40
L. Videokymographic analysis 40
M. Statistical analysis 41
Ⅲ. Results 42
A. Human VF-MSCs showed multi-potency of differentiation into various tissues 42
B. NTS does not induce cytotoxicity in VF-MSCs and increases cell viability 44
C. NTS increases migration of VF-MSCs and upregulates expression of MMP-2/MMP-9 45
D. NTS reduces fibrosis-related molecules and inhibits transformation of VF-MSCs into myofibroblasts 47
E. NTS modulates ECM deposition of VF mucosa and promotes recovery of VF function in vivo 49
Ⅳ. Discussion 55
Ⅴ. Conclusion 58
Ⅵ. References 59
국문요약 63
