Preparation and evaluation of naturally derived injectable hydrogels
- 주제(키워드) Injectable hydrogel
- 주제(DDC) 547
- 발행기관 아주대학교
- 지도교수 김문석
- 발행년도 2023
- 학위수여년월 2023. 8
- 학위명 박사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033008
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Drug delivery system (DDS) is a technology that optimizes drug treatment by preparing a formulation that can efficiently deliver the required amount of drug to the desired body site and minimize side effects. Over the past decades, DDS has been studied in various ways to solve the problems of existing formulations. Conventional drug delivery systems have unsolved problems such as rapid initial release, in vivo cytotoxicity, and immune rejection, and are not suitable for use in biomedical research. Therefore, advanced drug delivery systems are required to maintain plasma drug concentration in the therapeutic level, to release the contained drug released at the desired site, and not to have cytotoxicity and immune rejection. Among many DDS formulations, an injectable hydrogel can effectively deliver a drug to a desired area without surgery and induce a sustained release of the drug. In this study, an injectable hydrogel for increasing drug efficacy was prepared using natural biomaterials. Chapter 1 describes general introduction about background of the research, overall drug delivery systems and preparation of injectable hydrogel with naturally derived material. Chapter 2 describes the preparation of an injectable hydrogel with excellent biocompatibility using porcine cartilage-derived extracellular matrix, which is a natural biomaterial. Porcine cartilage-derived material has a limitation in preparing an injectable hydrogel due to its low hydrophilicity and a limitation in that it is biodegradable very quickly. Therefore, porcine cartilage-derived tissue was cross-linked using PEG having high biocompatibility and hydrophilicity. It was confirmed that the hydrogel crosslinked using PEG showed excellent biocompatibility in in vivo experiments and the physical properties and injectability were adjustable according to the crosslinking concentration of PEG. In Chapter 3, an injectable hydrogel was prepared using hyaluronic acid (HA), a biomaterial with excellent biocompatibility. Hyaluronic acid has the advantage of excellent biocompatibility but has the disadvantage of very rapid biodegradation. Due to this disadvantage, when used HA as a drug delivery system, it is difficult to suppress the initial burst of the drug loaded in HA and the duration of the drug effect is short. Therefore, in order to solve these disadvantages, we respectively introduced Tetrazine and trans cyclooctene into HA to occur cross-linking through Diels Alder click reaction and make it injectable in-situ forming hydrogel when directly into tumors. The HA hydrogel via the cross-linking reaction induced the sustained release of Doxorubicin, an anti-cancer drug, and increased the anti-cancer effect. In conclusion, the results of this study suggest methods for the preparation of injectable hydrogels using natural biomaterials.
more목차
CHAPTER 1. General introduction 1
1.1. Drug delivery systems 2
1.2. Hydrogels 4
1.3. Biomaterials 4
1.4. Natural materials 5
1.5. Synthetic materials 6
1.6. Strategy of this work 6
CHAPTER 2. Preparation and in vivo evaluation of an injectable crosslinked cartilage acellular matrix-PEG hydrogel scaffold derived from porcine cartilage 9
2.1. Introduction 10
2.2. Experimental section 14
2.2.1. Materials 14
2.2.2. Preparation of a CAM powder 15
2.2.3. Determination of Double Strand DNA of CAM Before and After Decellularization 16
2.2.4. Preparation of a Near Infrared (NIR) tagged CAM powder 17
2.2.5. Synthesis of COOH-PEG-400-COOH 17
2.2.6. Synthesis of NHS-PEG-400-NHS (PEG crosslinker) 18
2.2.7. Preparation of crosslinked CAM using PEG cross-linker 19
2.2.8. Preparation of CAM suspensions 20
2.2.9. Hydrophilicity test of CAM films and CAM hydrogel 21
2.2.10. Rheological properties of CAM hydrogels 22
2.2.11. Injectability test of CAM hydrogels 22
2.2.12. Animal experiment 23
2.2.13. In vivo biodegradation and biocompatibility experiments of NIR-labeled CAM, 0.6-CAM-GA, 0.6-CAM-PEG, 1-CAM-PEG, 3-CAM-PEG and 5-CAM-PEG hydrogels 24
2.2.14. In vivo implantation of CAM, 0.6-CAM-GA, and 0.6-CAM-PEG 24
2.2.15. Histological assay of CAM, 0.6-CAM-GA, and 0.6-CAM-PEG hydrogels 25
2.2.16. Statistical analysis 27
2.3. Results and Discussion 28
2.3.1. Preparation of CAM powder 28
2.3.2. Synthesis of COOH-PEG-400-COOH and NHS-PEG-400-NHS 31
2.3.3. Preparation of CAM, 0.6-CAM-GA, 0,6-CAM-PEG films 33
2.3.4. Preparation and characterization of CAM-PEG Powders 38
2.3.5. Hydrophilicity test of CAM, 0.6-CAM-GA, 0.6-CAM-PEG, 1-CAM-PEG, 3-CAM-PEG, and 5-CAM-PEG films 41
2.3.6. Rheological properties of CAM hydrogels 46
2.3.7. Injectability test of CAM hydrogels 49
2.3.8. In vivo biodegradation of NIR-labeled CAM, 0.6-CAM-GA, 0.6-CAM-PEG, 1-CAM-PEG, 3-CAM-PEG, and 5-CAM-PEG hydrogels 52
2.3.9. In vivo biodegradation of CAM, 0.6-CAM-GA, 0.6-CAM-PEG hydrogels 56
2.3.10. In vivo biocompatibility of CAM, 0.6-CAM-GA, 0.6-CAM-PEG hydrogels 58
2.4. Conclusion 62
CHAPTER 3. Anti-cancer activity of intratumorally injectable in-situ forming hyaluronic acid hydrogel 63
3.1. Introduction 64
3.2. Experimental section 69
3.2.1. Materials 69
3.2.2. Preparation of HA-Tet and HA-TCO hydrogels 69
3.2.3. Preparation of near-infrared (NIR) fluorescence-labeled HA-Tet (HA-Tet-NIR), HA-TCO (HA-TCO-NIR) and HA (HA-NIR) hydrogels 70
3.2.4. Preparation of HA-Dox Cx-HA-Dox, HA-NIR-Dox and Cx-HA-NIR-Dox formulation 71
3.2.5. Zeta potential of HA, HA-TCO, HA-Tet, Dox, HA-Dox, HA-TCO-Dox and HA-Tet-Dox 72
3.2.6. Rheological properties of hydrogels 72
3.2.7. Injectability test of hydrogels 73
3.2.8. In vitro release of Dox from HA and Cx-HA hydrogels 74
3.2.9. In vitro degradation of NIR tagged HA and Cx-HA hydrogels and Dox release from HA and Cx-HA hydrogels 75
3.2.10. In vitro anti-tumor activity 76
3.2.11. Inhibitiory effects 77
3.2.12. Animal study 77
3.2.13. Ex vivo fluorescent images of remained Dox and NIR labeled Cx-HA hydrogel within tumors and organs 78
3.2.14. Distribution of Dox in tumors after intratumoral injection 79
3.2.15. Histological assay 79
3.2.16. Statistical analysis 82
3.3. Result and discussion 83
3.3.1. Preparation and charaterization of Cx-HA hydrogel 83
3.3.2. Zeta potential of HA hydrogel formulations 86
3.3.3. Rheological properties and injectability of Cx-HA-Dox hydrogel formulation 90
3.3.4. In vitro Dox release and in vitro degradation of HA and Cx-HA hydrogels 95
3.3.5. In vivo Dox release and in vivo degradation of HA and Cx-HA hydrogels 99
3.3.6. In vivo anti-tumor effect of Dox of formulation 102
3.3.7. In vivo fluorescent images of remained Dox and NIR labelled Cx-HA hydrogel 105
3.3.8. Distribution of Dox in tumors after intra-tumoral injection 109
3.3.9. Histological assay 111
3.4. Discussion 117
3.5. Conclusion 121
CHAPTER 4. Overall conclusion 122
