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Formation of Multifunctional Hydrogels via Enzyme-mediated Crosslinking : 효소 매개 가교를 통한 다기능성 하이드로겔 제작

DC Field Value Language
dc.contributor.advisorHwang, Nathaniel Suk-Yeon-
dc.contributor.author김경민-
dc.date.accessioned2019-05-07T03:22:33Z-
dc.date.available2019-05-07T03:22:33Z-
dc.date.issued2019-02-
dc.identifier.other000000155766-
dc.identifier.urihttps://hdl.handle.net/10371/150843-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 공과대학 협동과정 바이오엔지니어링전공, 2019. 2. Hwang, Nathaniel Suk-Yeon.-
dc.description.abstractIn recent years, several attempts have been made to impart functionality to biomaterials in the field of tissue engineering. In particular, injectable form of hydrogels have many advantages such as ease of handling, in situ crosslinking, and being able to apply in 3D bioprinting applications. In this thesis, we demonstrate the synthesis of chitosan-based hydrogels containing Epigallocatechin gallate (EGCG) through recombinant tyrosinase from Streptomyces avermitilis (SA_Ty) that retains the anti-inflammation ability. Application of chitosan-EGCG (Chitosan-E) in wound healing application confirmed the regulation of the immune reaction due to the radical scavenging ability of hydrogel. Furthermore, we also present the synthesis of 3D printing hydrogel that can be crosslinked by SA-Ty by combining decellularized ECM and hyaluronic acid conjugated tyramine (HA_T).-
dc.description.abstract최근 조직공학 분야에서 다양한 재료에 기능성을 부여하는 시도들이 계속되고 있다. 특히 주사 가능한 하이드로겔은 상처 부위를 최소화할 수 있는 장점을 가지고 있어 꾸준히 연구되어 왔는데, 최근 들어 3D-프린팅 재료로서 사용될 수 있는 가능성 때문에 또다시 주목받고 있다. 우리는 녹차추출물인 Epigallocatechin gallate(EGCG)와 같은 기능성 분자 또는 다양한 생체신호 물질을 포함한 탈세포된 세포 외 기질을 이용해 주사 가능한 하이드로겔을 개발했다. Streptomyces avermitilis(SA_Ty)를 통해 EGCG를 산화시켜 제작한 키토산 기반 하이드로겔은 항 염증 능력을 가진다. 또한 상처 회복 모델을 통해 하이드로겔이 가지는 라디칼 소거 기능 때문에 면역반응이 조절되는 것을 확인했다. 또한 우리는 탈세포된 세포 외 기질과 Tyramine이 결합된 히알루론산을 적절히 배합하여 빠르게 가교 가능한 하이드로겔을 제작하였다. 종합하자면 우리가 제작한 하이드로겔은 상처 재생과 같은 질병에 효과적인 치료 효과를 보일 뿐 아니라 3D 프린팅 재료로서 사용 가능한 재료이다.-
dc.description.tableofcontentsTABLE OF CONTENTS

ABSTRACT ii
TABLE OF CONTENTS iv
CHAPTER ONE: TYROSINASE-MEDIATED CROSSLINKING OF POLYPHENOL FUNCTIONALIZED CHITOSAN FOR WOUND HEALING HYDROGEL １
2.1 Introduction １
2.2 Materials and methods ３
2.2.1 Synthesis of tyrosinase from Streptomyces avermitilis(SA-Ty) ３
2.2.2 Fabrication of chitosan hydrogel based on polyphenol ３
2.2.3 Rheological analysis of hydrogels ４
2.2.4 Measurement of Youngs modulus ４
2.2.5 Measurement of the swelling ratio of hydrogels ５
2.2.6 Scanning electron microscope(SEM) analysis ５
2.2.7 Measurement of EGCG releasing profile ６
2.2.8 Radical scavenging assays ６
2.2.9 Cell viability and proliferation test ７
2.2.10 In vitro, Anti-bacterial test ８
2.2.11 In vitro, Anti-inflammatory effects ８
2.2.12 In vivo, Wound healing model ９
2.2.13 Immunohistochemistry ９
2.2.14 Statistical analysis １０
2.3 Results １１
2.3.1 Synthesis and characterization of enzyme-mediated hydrogels １１
2.3.2 Mechanical and physical properties of hydrogels １２
2.3.3 Function of Chitosan-E hydrogels １２
2.3.4 In vivo, wound healing model １４
2.4 Discussion １５
CHAPTER TWO: SYNTHESIS OF DECELLULARIZED BRAIN ECM-BASED HYDROGEL FOR 3D-BIOPRINTING １８
3.1 Introduction １８
3.2 Materials and methods ２０
3.2.1 Materials ２０
3.2.2 Decellularization of porcine brain ２０
3.2.3 Synthesis of tyramine-modified hyaluronic acid(HA_T) ２１
3.2.4 Preparation of hydrogels ２２
3.2.5 Rheological analysis ２２
3.2.6 Swelling ratios analysis ２３
3.2.7 Scanning electron microscopy(SEM) analysis ２３
3.2.8 Measurement of Youngs modulus ２３
3.2.9 Statistical analysis ２４
3.3 Results & Discussion ２５
3.3.1 Synthesis and characterization of decellularized ECM-based hydrogel ２５
3.3.2 Mechanical and physical properties of hydrogels ２５
3.3.3 Further work ２６
REFERENCES ２７
초록(국문요약) ３０
LIST OF FIGURES AND TABLE ３１
Figures ３１
Figure 2.1 Three main component of tissue engineering ３１
Figure 2.2 The overall scheme of polyphenol-functionalized chitosan hydrogels formed via an enzyme reaction ３２
Figure 2.3 Mechanism of Chitosan-E hydrogels ３３
Figure 2.4 Sol fraction of Chitosan-E hydrogels ３４
Figure 2.5 Gelation of Chitosan-E hydrogels ３５
Figure 2.6 Rheological analysis of Chitosan-E hydrogels ３６
Figure 2.7 Mechanical properties of Chitosan-E hydrogels ３７
Figure 2.8 Microstructure of Chitosan-E hydrogels ３８
Figure 2.9 Releasing profile and function of hydrogel ３９
Figure 2.10 Scheme of anti-bacterial test ４０
Figure 2.11 Anti-bacterial ability of hydrogel ４１
Figure 2.12 Biocompatibility of hydrogel ４２
Figure 2.13 Inflammation modulation ability of hydrogel ４３
Figure 2.14 Chitosan-E hydrogels injection model ４４
Figure 2.15 Scheme of wound healing model and wound healing process ４５
Figure 2.16 Wound closing images and ratio ４６
Figure 2.17 Histology images ４７
Figure 2.18 Immunohistochemistry of wound healing model ４８
Figure 2.19 In vivo, vascularization in the wound area ４９
Figure 3.1 Scheme for the synthesis of decellularized ECM-based hydrogel ５０
Figure 3.2 Decellularization step ５１
Figure 3.3 Characterization of decellularized ECM ５２
Figure 3.4 Gelation images of decellularized ECM hydrogel ５３
Figure 3.5 Rheological analysis of decellularized ECM hydrogel ５４
Figure 3.6 Mechanical properties of decellularized ECM-based hydrogel ５５
Figure 3.7 Scanning electron microscope image ５６
Figure 3.8 Further work ５７
Tables ５８
Table 3.1 DNA quantification ５８
Table 3.2 Youngs modulus of decellularized ECM-based hydrogel ５９		-
dc.language.isoeng-
dc.publisher서울대학교 대학원-
dc.subject.ddc660.6-
dc.titleFormation of Multifunctional Hydrogels via Enzyme-mediated Crosslinking-
dc.title.alternative효소 매개 가교를 통한 다기능성 하이드로겔 제작-
dc.typeThesis-
dc.typeDissertation-
dc.contributor.AlternativeAuthorKyungmin Kim-
dc.description.degreeMaster-
dc.contributor.affiliation공과대학 협동과정 바이오엔지니어링전공-
dc.date.awarded2019-02-
dc.identifier.uciI804:11032-000000155766-
dc.identifier.holdings000000000026▲000000000039▲000000155766▲-
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