Artificial Cell Membrane Models with Extracellular Matrix, Cytoskeletal Proteins, and the Applications
- 주제어 (키워드) artificial cell , giant unilamellar vesicle , synthetic biology , wound healing
- 발행기관 서강대학교 일반대학원
- 지도교수 신관우
- 발행년도 2024
- 학위수여년월 2024. 8
- 학위명 박사
- 학과 및 전공 일반대학원 화학과
- 실제 URI http://www.dcollection.net/handler/sogang/000000078856
- UCI I804:11029-000000078856
- 본문언어 영어
- 저작권 서강대학교 논문은 저작권 보호를 받습니다.
초록 (요약문)
Although creating living artificial cells remains challenging, researchers are advancing by reconstructing Giant Unilamellar Vesicles (GUVs), promising platforms for designing more functional artificial cells. Therefore, understanding lipid membrane-protein interactions is essential for successfully forming artificial cells and engineering synthetic biological systems, including drug delivery systems. This thesis explores the intricate interactions between cell membranes and actin cytoskeletal, extracellular matrix (ECM) proteins, as well as their applications in biomedical engineering and biotechnology. An actin-GUV system designed to maintain membrane integrity during adhesion to solid surfaces and prevent membrane rupture under high tension by impeding the expansion of membrane pores is presented. This research further investigates lipid redistribution on phase-separated GUVs tailored for selective ECM protein binding, emphasizing the significant impact of ECM proteins on membrane lipid rearrangement and lipid lateral diffusion. Additionally, the study highlights the role of negatively charged small unilamellar vesicles (SUVs) in effectively unfolding fibronectin (FN) before delivery, thereby enhancing tissue regeneration in vitro and in murine models. Collectively, these findings provide valuable insights into the dynamics of ECM and actin protein interactions with cell membranes and underscore the potential applications of artificial cell systems in advancing medical and biotechnological fields.
more목차
Chapter 1. Reseach background and Motivation 1
1.1. Artificial cells and their applications 1
1.2. Dissertation organization and significance 3
Chaper 2 Enhancement of membrane integrity by ATP-activated actin in artificial cells 5
2.1. Introduction 5
2.2. Materials and methods 7
2.3. Results 11
2.3.1. Actin polymerization triggered by divalent cations 11
2.3.2. Preservation of damaged GUVs by actin meshwork formation 13
2.3.3. The morphology of actin-GUV rescued by actin meshwork 16
2.3.4. The permeability of actin-GUV to small molecules 18
2.4. Discussion and conclusion 19
2.5. Conclusion 26
Chapter 3 Lipid domain reorganization in artificial cell model with extracellular matrix proteins 26
3.1. Introduction 26
3.2. Materials and Methods 29
3.3. Results 32
3.3.1. Binding to the ECM changes the distribution of liquid disorder (Ld) lipid domains 32
3.3.2. Intrinsic factors affect the topographical changes of the Ld phase 35
3.3.3. The binding of ECM stabilized the Ld domain by impeding lateral diffusion 38
3.4. Discussion 41
3.5. Conclusion 45
Chapter 4 Efficient fibronectin delivery via liposomes for enhanced tissue regeneration 47
4.1. Introduction 47
4.2. Materials and methods 50
4.3. Results 57
4.3.1. FN experienced stretching upon interaction with negatively charged liposomes 57
4.3.2. FN-SUVs enhanced cell attachment and proliferation more effectively than soluble FN 61
4.3.3. Enhanced cell migration through SUV-FN delivery 65
4.3.4. FN-SUV demonstrated superior improvement in wound healing in rats with ulcerative colitis compared to FN in its compact form 68
4.4. Discussion 72
4.5. Conclusion 74
4.6. Supplementary 75
References 86
