신규 조직공학적 성대주입물의 성대재생 효과분석
The Analysis of Novel Injectable Tissue - Engineered Materials for Vocal Fold Regeneration
- 주제(키워드) Injection laryngoplasty , Vocal fold palsy , Vocal fold scar , Hyaluronic acid , hepatocyte growth factor
- 발행기관 아주대학교
- 지도교수 김철호
- 발행년도 2017
- 학위수여년월 2017. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000025652
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Injection laryngoplasty (IL) is one of the most suitable options for treatment of glottic insufficiency, which is caused by vocal fold (VF) paralysis, atrophy, or scarring. The purpose of this study is to demonstrate favorable VF augmentation and regeneration by using a several material such as hyaluronic acid, MPEG-b-(PCL-ran-PLLA) diblock copolymers (C97L3; CL/LA ratio 97:3) incorporating hepatocyte growth factor (HGF). Hyaluronic acid (HA) is a widely used material for VF injection. We intended to evaluate the durability and efficiency of HA of different particle sizes for VF augmentation. An in situ-forming gel system comprised of diblock copolymer formed from polyethylene glycol (PEG) and polycaprolactone (PCL) could be used in controlled drug delivery for tissue remodeling. Gradual release of HGF from C97L3 is detected and biochemical properties of released HGF are maintained. After six and ten weeks, the injected site was evaluated endoscopically, histologically, radiologically, and functionally. None of the 24 rabbits showed any signs of respiratory distress. Computed tomography (CT) images and endoscopic evaluation revealed sufficient augmented volume of the injected VF in all treated groups 6 weeks after the injection. Histological data at week 10 showed that unequal particle-sized HA did not migrate from its original injection site, while other HAs migrated into the arytenoid cartilage. Videokymographic analysis showed more favorable vibrations of unequal particle-sized HA injected VF mucosa 10 weeks post-injection, compared to the other treatment groups. A scar is made with microscissors on both VFs in 32 rabbits, followed by injection of HGF-only, C97L3-only, or HGF-C97L3 composite gel in the left side VF, while the right side VF is left untreated. In vivo fluorescence live imaging system demonstrates that C97L3 enables the sustained release of injected HGF in the scarred VF for 12 weeks. The histological analysis shows increased glycosaminoglycan including hyaluronic acid accumulation and decreased collagen deposition. Videokymographic analysis shows more favorable vibrations of HGF-C97L3 treated VF mucosa, compared to other treatment groups. In conclusion, the controlled HGF release helps to regulate extracellular matrix synthesis, and leads to the eventual functional improvement of the scarred VF.
more목차
Part I. Efficiency and durability of hyaluronic acid of different particle sizes as an injectable material for VF augmentation 1
Introduction 2
Materials and Methods 4
A. Injection materials 4
B. Animal and surgical procedure 6
C. Endoscopic and radiologic assessment 8
D. Videokymographic analysis 8
E. Histologic assessment 9
F. Statistical Analysis 9
Results 10
A. Assessment of in vivo autologous injection material 10
B. Histological assessments of injection material 13
C. Videokymographic analysis 15
Discussion 17
Part II. Controlled Release of Hepatocyte Growth Factor from MPEG-b-(PCL-ran-PLLA) Diblock Copolymer for Improved Vocal Fold Regeneration 21
Introduction 22
Materials and Methods 25
A. Preparation of MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 25
B. In Vitro Release Profile of HGF from MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 30
C. Fluorescence Labeling and Tracing and Quantitative Analysis of HGF in MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 32
D. Animal Surgical Procedures 32
E. Tracing and Quantitative Analysis of Fluorescence-Labeled HGF 33
F. Endoscopic and Histologic Analysis 33
G. Videokymographic analysis 34
H. Statistical Analysis 34
Results 35
A. Controlled In Vitro Release of HGF from MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 35
B. Biochemical Properties of Released HGF from MPEG-b-(PCLran-PLLA) Diblock Copolymers 36
C. In Vivo Tracing of HGF Incorporated with MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 39
D. Morphologic and Structural Regeneration of VF by Sustained Released HGF from MPEG-b-(PCL-ran-PLLA) Diblock Copolymers 42
E. Enhanced Vibratory Function in HGF-MPEG-b-(PCL-ran-PLLA) Diblock Copolymer Treated Vocal Folds 46
Discussion 48
Reference 53
국문 요약 59
