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Fabrication of photo-crosslinked and RGD modified ECM-based hydrogels for functional cartilage tissue engineering
연골 조직 공학을 위한 아르지닌-글리신-아스파르트산 펩타이드와 세포외기질 기반의 광중합성 하이드로겔 합성

DC Field Value Language
dc.contributor.advisor황석연-
dc.contributor.author김환-
dc.date.accessioned2017-07-17T08:45:51Z-
dc.date.available2017-07-17T08:45:51Z-
dc.date.issued2014-08-
dc.identifier.other000000020720-
dc.identifier.urihttps://hdl.handle.net/10371/127103-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 화학생물공학부, 2014. 8. 황석연.-
dc.description.abstractArticular cartilage damage is a persistent and increasing problem with an aging population. Though it is not life threatening, damages do strongly affect ones quality of life. Such damages can be resulted from a variety of causes, such as traumatic accident or wear and tear over time. Articular cartilage does not easily go through the regeneration step after having injuries or disease leading to loss of tissue and formation of a defect. Therefore, strategies to achieve complete cartilage restoration remain a challenge. Photopolymerizing-based injectable hydrogels have long received attention in the cartilage tissue engineering, due to their unique biocompatibility, flexible method of synthesis, range of constituents, and desirable physical characteristics. In the present study, we have introduced unique bioactivity within the photopolymerizing hydrogels by copolymerizing polyethylene glycol diacrylate (PEGDA) macromers with various photo-crosslinkable extracellular matrix (ECM) molecules (hyaluronic acid and chondroitin sulfate) and integrin binding peptides (RGD peptide). Results indicate that the cellular morphology as observed by the actin cytoskeleton structures, was strongly dependent on the types ECM components as well as the presence of integrin binding moieties. Furthermore, chondroitin sulfate (CS)-based hydrogel with integrin binding moieties increased the lubricin (or known as superficial zone protein, SZP) gene expression by the encapsulated chondrocytes. Additionally, CS-based hydrogel with RGD peptide increased DNA, GAG, and collagen contents compared HA-based gel and non-modified control hydrogels. This study demonstrates that integrin-mediated ECM microenvironments should be considered for the hydrogel-based cartilage tissue engineering applications.-
dc.description.abstract관절연골의 손상은 인구의 고령화와 함께 지속적으로 증가하는 경향성을 보이고 있다. 그러나, 현재까지 연골의 전체적 재생 및 최소한의 기능 회복에 대한 치료법은 여전히 확립되어 있지 않다. 광중합 기반의 생체주입식 하이드로겔은 선행연구로부터 생체 적합성과 합성과정의 유동성, 구성 요소의 범위 및 물리적 특성의 이유로 연골조직공학에 적합한 재료로 인식되어왔다. 본 연구에서는 다양한 기능성의 생물작용을 내포한 광중합 기반의 하이드로겔을 폴리에틸렌 글리콜 디아크릴레이트 (이하, PEGDA), 세포외기질 (이하, ECM) 및 아르지닌-글리신-아스파르트산 펩타이드 (이하, RGD 펩타이드)와 함께 가교하여, 하이드로겔 내에서 고유한 생리 활성을 도입 및 유지 할 수 있도록 하였다. 따라서, 세포의 액틴 골격구조를 유심히 살펴 본 결과, ECM 및 RGD 펩타이드 존재 유무에 따라 세포의 형태가 주변 미세환경에 의존한다는 것이 밝혀졌다. 덧붙혀서, 여러 종류의 하이드로겔중, ECM의 한 종류인 황산 콘드로이틴 (Chondroitin Sulfate, 이하 CS) 기반과 RGD 펩타이드가 혼합된 하이드로겔에서 소 연골세포의 가장 높은Lubricin 의 유전자 증가를 보여주었다. 또한, 위에서 제시한 CS 기반의 하이드로겔 은 다른 ECM 의 일종인 히알루론산 (Hyaluronic Acid, 이하 HA) 기반의 하이드로겔과 비교하여, 소 연골세포의 유전자 (DNA) 와, 글리코사미노글리캔 (Glycosaminoglycan, Gags) 및 콜라겐의 양이 증가 하였다. 이 연구 결과는 RGD 펩타이트와 ECM의 미세 환경의 두 인자가 하이드로겔 기반 연골 조직 공학 응용을 위해 반드시 고려 되어야 함을 보여준다.-
dc.description.tableofcontentsTABLE OF CONTENTS

ABSTRACT 1
TABLE OF CONTENTS 3
LIST OF FIGURES AND TABLES 5

TABLE OF CONTENTS

ABSTRACT 1
TABLE OF CONTENTS 3
LIST OF FIGURES AND TABLES 5

CHAPTER ONE: THE SCIENTIFIC BACKGROUND AND RESEARCH PROGRESS 6

1.1 Overview 6
1.2 Polyethylene glycol diacrylate (PEGDA) hydrogel 7
1.3 RGD peptide and extracellular matrix (ECM) components 8
1.4 Cartilage tissue engineering 10
1.5 Research aims 11

CHAPTER TWO: SYNTHESIS OF CELL RESPONSIVE ECM-BASED HYRDROGELS 13

2.1 Introduction 13
2.2 Materials and methods 13
2.2.1 Isolation and culture of bovine chondrocyte 13
2.2.2 Methacrylation of chondroitin sulfate and hyaluronic acid 14
2.2.3 Photoencapsulation 15
2.2.4 Swelling properties and mechanical tests 18
2.2.5 Statistical analysis 18
2.3 Results 19
2.3.1 Design and synthesis of photo-crosslinkable biomacromolecules for RGD/RDG -modified ECM-based hydrogels 19
2.3.2 Characterization of RGD/RDG-modified ECM-based hydrogels 24
2.4 Discussion 27

CHAPTER THREE: TISSUE ENGINEERING APPLICATION OF CELL RESPONSIVE ECM-BASED HYRDROGELS 29

3.1 Introduction 29
3.2 Materials and methods 29
3.2.1 Cellular viability and morphological analysis 29
3.2.2 Biochemical analysis 30
3.2.3 Histological and immunofluorescence analysis 31
3.2.4 Real-Time PCR 32
3.2.5 Statistical analysis 32
3.3 Results 33
3.3.1 Viability of bovine chondrocytes 33
3.3.2 Effect of ECM-based hydrogels on chondrocyte morphology, proliferation, and ECM synthesis 35
3.3.3 Histological and immunostaining analysis of engineered cartilage tissues 39
3.3.4 Gene expression analysis of chondrocytes in RGD/RDG-modified ECM-based hydrogels 44
3.4 Discussion 46

CONCLUSION 49

REFERENCE 50

초록(국문요약) 57

감사의 글 59
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dc.formatapplication/pdf-
dc.format.extent3050613 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectHydrogel-
dc.subjectBovine Chondrocyte-
dc.subjectSZP-
dc.subjectRGD-
dc.subjectChondroitin Sulfate-
dc.subjectHyaluronic Acid-
dc.subjectCartilage Tissue Engineering-
dc.subject.ddc660-
dc.titleFabrication of photo-crosslinked and RGD modified ECM-based hydrogels for functional cartilage tissue engineering-
dc.title.alternative연골 조직 공학을 위한 아르지닌-글리신-아스파르트산 펩타이드와 세포외기질 기반의 광중합성 하이드로겔 합성-
dc.typeThesis-
dc.contributor.AlternativeAuthorHwan Kim-
dc.description.degreeMaster-
dc.citation.pages58-
dc.contributor.affiliation공과대학 화학생물공학부-
dc.date.awarded2014-08-
Appears in Collections:
College of Engineering/Engineering Practice School (공과대학/대학원)Dept. of Chemical and Biological Engineering (화학생물공학부)Theses (Master's Degree_화학생물공학부)
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