혈관 내피세포화 촉진을 위해 효소를 이용한 RGD 고정화 폴리우레탄 표면 개질
Enzyme-mediated in situ surface grafting of cellular adhesion ligand onto polyurethane for improved endothelialization
- 주제(키워드) polyurethane , RGD , tyrosinase , surface modification
- 발행기관 아주대학교
- 지도교수 박기동 교수님
- 발행년도 2012
- 학위수여년월 2012. 2
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000012386
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The Arg-Gly-Asp (RGD) sequence is one of the most effective cellular adhesion ligands and has been used to improve cellular activity of biomaterials for tissue regenerative medicine. Aim of this study was to develop new facile immobilization methods of RGD on electrospun polyurethane (PU) meshes using tyrosinase (Ty) to promote attachment and proliferation of human umbilical vein endothelial cells (HUVECs). The PRGDGGGGGY (PRGD-Y) peptides were readily immobilized on PU substrates for 1 h using 0.4 KU/mL of Ty. Water contact angles and X-ray photoelectron spectroscopy (XPS) was used to verify that the PRGD-Y peptide was effectively conjugated on the PU (0.120 nmol/mgmesh) and the surface concentration was shown to be proportional to the peptide feed concentration. Enhanced HUVEC attachment on the polymer substrate was observed after 1 day and the immobilized PRGD-Y was clearly demonstrated to promote proliferation and spreading of the cells on the surface, indicating that the method developed for the conjugation of the peptides on the PU was very efficient. These results suggest that this enzyme-triggered immobilization method for the cellular adhesion ligand, RGD, onto the PU mashes may be an efficient tool to improve the biological activity of substrates especially for vascular tissue engineering.
more목차
I. INTRODUCTION 1
A. Artificial vascular graft 1
B. Preparation of scaffold applied to artificial vascular grafts 3
1. Biomaterials for ideal scaffolds 3
2. Polyurethane 5
C. Electro-spinning 8
D. Surface modification for endothelialization 11
1. RGD of bioactive molecule for endothelialization 11
2. Tyrosinase for enzyme-oxidative reaction 14
E. Objective 17
II. EXPERIMENTAL PARTS 18
A. Materials 18
B. Fabrication of PU mesh 18
C. Preparation of micro-fibrous PU mesh and immobilization of PRGD-Y on the mesh 19
D. Surface characterization of the PRGDY-PU meshes 19
1. SEM 20
2. XPS 20
3. Contact angle measurement 20
E. Quantification of concentration of the PRGD-Y on the PU meshes 22
1. Flourescamine assay 22
F. In vitro cell study using human umbilical vein endothelial cells (HUVECs) 23
III. RESULTS AND DISCUSSION 24
A. Characterizations of PU mesh and PRGD-Y immobilized PU mesh 24
1. SEM images and contact angle results of PU mesh and PRGD-Y immobilized PU mesh 24
2. Atomic compositions of PU surfaces 27
B. Amount of the immobilized PRGD-Y on PU meshes 29
C. In vitro cell attachment and proliferation HUVEC on the PU meshes 31
IV. CONCLUSIONS 35
V. REFERENCES 36
