온도감응성 키토산-플루로닉 하이드로젤을 이용한 연골 재생에 관한 연구
Thermosensitive chitosan-Pluronic hydrogels for the cartilage regeneration
- 주제(키워드) 연골 재생 , 플루로닉 하이드로젤 , 온도감응성 키토산
- 발행기관 아주대학교 대학원
- 지도교수 Park, Ki Dong
- 발행년도 2008
- 학위수여년월 2008. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의용공학과정
- 본문언어 영어
초록/요약
Chitosan-based hydrogel is a good candidate for the cartilage regeneration, since it shares some characteristics with a variety of glucosaminoglycan (GAG) which is one of the major macromolecules found in the articular cartilage. In this study, we designed thermosensitive chitosan-Pluronic (CP) hydrogel as a cell supported scaffold for the cartilage regeneration. The CP copolymer was synthesized by grafting Pluronic onto chitosan using EDC/NHS chemistry. Sol-gel phase transition and mechanical properties of the CP hydrogel were investigated by rheological experiments. The CP hydrogel showed the sol-gel transition around 37°C and the storage modulus (G′) was over 10⁴ kPa, showing potentials of an injectable hydrogel for the cartilage regeneration. The CP hydrogel was rapidly formed by increasing temperature. Morphologies of freeze dried CP hydrogels were observed by SEM. In addition, in vitro growth factor release study was performed using bone morphogenic protein (BMP-7). BMP-7 was released sustainedly from the CP hydrogels for 20 days. Furthermore, in vitro cell culture was performed using bovine chondrocyte. The proliferation of bovine chondrocyte and amount of synthesized GAG were increased for 28 days. Therefore, the obtained results suggest that the CP hydrogel has a potential for cartilage regeneration and it can be also useful as a new injectable biomaterial for tissue engineering.
more목차
Ⅰ. INTRODUCTIONS = 1
A. Theoretical background = 1
1. Chitin and chitosan = 1
2. Hydrogels = 5
3. Thermosensitive hydrogels = 9
4. Cartilage injury and repair = 12
B. Objectives = 15
Ⅱ. EXPERIMENTS = 16
A. Materials = 16
B. Synthesis of thermosensitive Chitosan-Pluronic copolymer = 16
C. Critical aggregate concentration (CAC) analisys = 19
D. Preparation of the Chitosan-Pluronic hydrogels = 20
E. Characterizations of the Chitosan-Pluronic hydrogels = 20
1. Rheological anaysis = 20
2. Swelling ratio and degradation behavior = 20
3. Morphology = 21
E. In vitro growth factor release study = 21
F. In vitro cell culture using bovine chondrocyte = 23
1. Isolation and expansion of chondrocyte = 23
2. Chondrocyte encapsulation and culture = 23
3. Cell viability test = 25
4. Microscopic observation = 25
5. Glycosaminoglycan (GAG) content = 25
6. RNA extraction and reverse transcription-polymerase chain reaction analysis = 26
7. Statistical analysis = 26
Ⅲ. RESULTS AND DISCUSSION = 27
A. Characterizations of the chitosan-Pluronic hydrogels = 27
1. Critical aggregate concentration (CAC) = 27
2. Shear modulus = 30
3. Swelling ratio and degradation time assay = 32
4. Morphology = 33
B. In vitro growth factor release = 34
C. In vitro cell culture = 36
1. Viability and proliferation of chondrocyte = 36
2. Synthesis of glycosaminoglycan (GAG) = 39
3. Gene expression of extracellular matrix molecules = 40
Ⅳ. CONCLUSIONS = 42
V. REFFERENCES = 43
