연골 세포외기질로 만들어진 막의 항유착성 평가
Anti-adhesive properties of biofilm made of cartilage extracellular matrix
- 주제(키워드) Chondrocyte-derived extracellular matrix , anti-angiogenesis , anti-adhesion , surface modification , postsurgical adhesion
- 발행기관 아주대학교
- 지도교수 민병현
- 발행년도 2016
- 학위수여년월 2016. 2
- 학위명 박사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000021748
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Adhesion is a phenomenon that takes place when the balance between fibrin deposition and fibrinolysis breaks with chronic inflammation during the wound healing process. Different tissue places are connected between abnormal fibrous bands of scar tissue. While the pathogenesis of adhesion is still unclear, the main mechanism is thought to be inflammatory reactions that stimulate mesothelial layer of tissue. Severe adhesion can hinder physiological activity of involved organs, leading to secondary diseases such as bowel obstruction, pelvic pain, chronic abdominal pain and infertility. Modalities treating these adhesions induce microsurgery, fibrinolytic agents, anticoagulants, anti-inflammatory agents, and drugs such as antibiotics. The most common modality is directly separating adhered tissues using a bioreabsorbable physical barrier. Most barriers, however, act as physical barriers without biological effects. In case of natural polymer derived barriers, it is rapidly degraded before the end of the healing process. Barriers composed of synthetic polymers have the limitation of requiring implant removal. Although the cross-linking of the natural material and drug release systems has been explored to overcome limitation of previous materials, they still fall short of clinically preventing adhesion. In previous studies, we confirmed that the cartilage extracellular matrix is a biocompatible and antiangiogenic functional, and available as a physical barrier to block the movement of the cells. In this study, we expected that extracellular matrix membrane can suppress the postsurgical adhesions by inhibiting the cell adhesion through the surface modification CHAPTER I: In this experiment, we saw how the cartilage extracellular matrix (CEM) affects the activity of the endothelial cells during angiogenesis in vitro. Cartilage extracellular matrix (CEM) inhibited endothelial cells (ECs) adhesion when compared to small intestinal submucosa (SIS). Concentration dependent inhibition of EC attachment was observed in the CEM, while EC attachment and cytoskeleton formation was observed in all concentration in the SIS group. Decrease in cell proliferation was also observed in the CEM group. CEM also interfered with tube formation when compared to SIS during matrigel plug assay. The mode of action during inhibition of tube formation was not due to physical properties of the powder, but rather due to biological component of CEM such as chondroitin sulfate. CHAPTER II: The surface of membrane was modified with PLL coating (PLL 10), thereby neutralizing its surface charge. The PLL10 CEM suppressed fibroblast and endothelial cell adhesion in vitro and moreover, inhibited abdominal adhesion in vivo. This study demonstrates that PLL10 surface modification renders the charge of neutral polymer extracellular matrix surface, thereby inhibiting cell and tissue adhesion. Furthermore, this study suggests a means to modify extracellular matrix surfaces to meet specific requirements of target tissue in preventing post-surgical adhesions.
more목차
General Introduction- 1
1.Postsurgical adhesion 1
2.Pathogenesis of adhesions 1
2.1General wound healing 2
2.1.1Cellular events in wound healing process 2
2.1.2The role of angiogenesis in wound healing process 3
2.2Types of abnormal adhesions 4
2.2.1Abdominal adhesion 4
2.2.2Tendon adhesion 4
2.2.3Guided tissue regeneration (GTR) 5
3.Prevention of anti-adhesion 6
3.1Pharmacological approaches 7
3.2Physical barrier approaches 7
3.2.1Components of the anti-adhesion agent 7
3.2.2Gel type 8
3.2.3Film type 9
3.3Drug releasing physical barriers 9
3.4Products of anti-adhesion agents 11
3.4.1Solution/Gel type 11
3.4.2Film type 11
4.Interaction of materials and cells 13
4.1Cell adhesion in postsurgical adhesion 13
4.2Cell adhesion on biomaterial 13
5.Cartilage extracellular matrix 15
5.1Tissue engineering application in previous studies 15
5.2Anti angiogenic study in previous studies 16
6.The aims of this study 17
Chapter I. 18
1.Introduction 18
2.Materials and Methods 20
2.1Cell culture 20
2.2Preparation of SIS and CEM powder 20
2.3Adhesion and proliferation assay for fibroblast and HUVECs 20
2.4Tube formation assay 21
2.5Statistical analysis 21
3.Results 22
3.1Adhesion of HUVECS on SIS and CEM powder 22
3.2Proliferation and cytotoxicity of HUVECs on SIS and CEM powder- 26
3.3Tube formation effects of soluble molecules from SIS and CEM 29
3.4Tube formation effects of SIS and CEM powder 31
3.5Tube formation effects of component of SIS and CEM 35
4.Discussion 37
Chapter II. 40
1.Introduction 40
2.Materials and Methods 41
2.1Preparation of chondrocyte derived extracellular matrix and modification 41
2.2Characterization of CEM and PLL10 42
2.3Attached cell morphology and cell 42
2.4Subcutaneous adhesion evaluation in mouse model of CEM and PLL10 43
2.5Immunohistochemical analysis 43
2.6Cecum adhesion in rat mode of CEM and PLL10 44
2.7Statistical analysis 44
3.Results 45
3.1Surface characterization of CEM and PLL film 45
3.2In vitro cell morphology and proliferation on CEM and PLL film 49
3.3Prevention of subcutaneous adhesions 51
3.4Prevention of cecum adhesions 57
4.Discussion 59
Summary & Conclusion of Part 64
Further study 65
References 67
국문요약 84
List of publications 86
