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Riboflavin induced photo-cross-linking of collagen hydrogel and its application in tissue engineering : 라이보플라빈을 통한 콜라겐 하이드로겔의 광가교결합 및 조직공학에의 응용

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dc.contributor.advisorNathaniel S. Hwang-
dc.contributor.author허지승-
dc.date.accessioned2017-07-19T05:54:37Z-
dc.date.available2017-07-19T05:54:37Z-
dc.date.issued2015-02-
dc.identifier.other000000025818-
dc.identifier.urihttps://hdl.handle.net/10371/129370-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 화학생물공학부, 2015. 2. Nathaniel S. Hwang.-
dc.description.abstractA meniscus tear is a common knee injury but its regeneration remains a clinical challenge. Recently, collagen-based scaffolds have been applied in meniscus tissue engineering. Despite its prevalence, application of natural collagen scaffold in clinical setting is limited due to its extremely low stiffness and rapid degradation. The purpose of the present study was to increase the mechanical properties and delay degradation rate of collagen-based scaffold by photo-crosslinking using riboflavin and UV exposure. Riboflavin is a biocompatible vitamin B2 that showed minimal cytotoxicity compared to conventionally utilized photo-initiator. Furthermore, collagen photo-cross-linking with riboflavin improved mechanical properties and delayed enzymatic degradation rate of collagen scaffolds. Furthermore, in order to observe the meniscus tissue engineering, fibrochondrocytes were encapsulated. Photo-crosslinking of collagen resulted in reduced contractility and enhanced gene expression levels for the collagen II and aggrecan. Additionally, for activation of meniscus regeneration, hyaluronic acid (HA) was incorporated with collagen gel and showed increase of meniscus related gene level for the type I collagen, type II collagen, and aggrecan. Based upon these results, we demonstrate that photo-cross-linked collagen HA have potential for utilization in the scaffold-based meniscus tissue engineering.-
dc.description.tableofcontentsAbstract 1
Contents 3
List of Figures 6
Chapter 1. THE SCIENTIFIC BACKGROUND AND RESEARCHS PROGRESS 7
1.1 Meniscus regeneration 7
1.2 Collagen as a biomaterial 9
1.3 Collagen crosslinking with riboflavin 9
1.4 Research aims 10
Chapter 2. PHOTO-CROSS-LINKING OF COLLAGEN HYDROGEL WITH RIBOFLAVIN 12
2.1 Introduction 12
2.2 Experimental 12
2.2.1 Materials 12
2.2.2 Fibrochondrocytes isolation and culture 13
2.2.3 Photo-cross-linking of collagen gel and cell encapsulation 13
2.2.4 CD measurement 14
2.2.5 Rheological property, swelling ratio, in vitro degradation 14
2.2.6 Swelling ratio 14
2.2.7 Live/dead assay and contraction assay 15
2.2.8 Degradation 15
2.2.9 Real time-PCR analysis 15
2.2.10 Histological analysis 16
2.2.11 Statistical analysis 16
2.3 Results 18
2.3.1 Preparation of photo-cross-linked hydrogel 18
2.3.2 Dose dependent elastic modulus of hydrogel 20
2.3.3 Characterization of treatment time dependent photo-cross-linked collagen gel 22
2.3.4 Contraction assay and degradation rate analysis 28
2.3.5 Gene expression analysis and histological staining of fibrochondrocytes in COL and COL-RF 31
2.3.6 Histological staining of fibrochondrocytes in COL and COL-RF 33
2.4 Discussion 35
Chapter 3. PHOTO-CROSS-LINKED COLLAGEN GEL WITH HYALURONIC ACID 38
3.1 Introduction 38
3.2 Experimental 38
3.2.1 Material 38
3.2.2 Synthesis of cross-linked HA 39
3.2.3 Photo-cross-linking of collagen gel and cell encapsulation 39
3.2.4 Carbazole assay 40
3.2.5 Fourier-Transformed Infra-Red Spectroscopy (FT-IR) 40
3.2.6 Real time-PCR analysis 40
3.2.7 Subcutaneous implantation of hydrogel 41
3.2.8 Histological staining 41
3.2.9 Statistical analysis 41
3.3 Results 42
3.3.1 Synthesis of cross-linked HA 42
3.3.2 HA retention within hydrogel 44
3.3.3 Gene expression analysis of fibrochondrocytes in collagen hydrogels. 46
3.3.4 In vivo meniscus regeneration 48
3.4 Discussion 50
Chapter 4. Conclusions 52
국 문 초 록 58		-
dc.formatapplication/pdf-
dc.format.extent2226581 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjecthydrogel-
dc.subject.ddc660-
dc.titleRiboflavin induced photo-cross-linking of collagen hydrogel and its application in tissue engineering-
dc.title.alternative라이보플라빈을 통한 콜라겐 하이드로겔의 광가교결합 및 조직공학에의 응용-
dc.typeThesis-
dc.contributor.AlternativeAuthorHeo Jiseung-
dc.description.degreeMaster-
dc.citation.pages59-
dc.contributor.affiliation공과대학 화학생물공학부-
dc.date.awarded2015-02-
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