Design and evaluation of hyaluronic acid-oleic acid nanoparticles for boosting therapeutic drug delivery
- 주제(키워드) Hyaluronic acid , oleic acid , nanoparticles , cyclosporine A , ocular delivery , doxorubicin , anticancer
- 주제(DDC) 615.1
- 발행기관 아주대학교
- 지도교수 Beom-Jin Lee
- 발행년도 2023
- 학위수여년월 2023. 8
- 학위명 박사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000032872
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
목차
Chapter I. Self-assembled nanodrops of hyaluronic acid-oleic acid conjugates for improving the ocular delivery of cyclosporine A 1
1. Introduction 1
2. Materials and Methods 4
2.1. Materials 4
2.2. Synthesis of oleic acid-hyaluronic acid conjugates (HOCs) 5
2.2.1. Synthesis of cystamine-modified hyaluronic acid (HA-CYS) 5
2.2.2. Synthesis of oleic acid-conjugated HA-CYS (HOCs) 5
2.3. Preparation of CsA-loaded self-assembled HONs (CsA-HONs) 6
2.4. Physicochemical characterizations of HOCs and CsA-HONs 7
2.4.1. Hydrogen-1 nuclear magnetic resonance (1H-NMR) 7
2.4.2. UV-VIS absorbance measurement 7
2.4.3. FTIR analysis 7
2.4.4. Quantification of free amine groups by TNBS method 7
2.4.5. CMC determination 8
2.4.6. PXRD analysis 9
2.4.7. DLS measurement 9
2.4.8. Morphology of CsA-HONs 9
2.4.9. HPLC analysis 10
2.4.10. Measurement of absorbance 10
2.5. In vitro drug release study 10
2.6. In vitro drug permeation study 11
2.7. In vitro cytocompatibility of CsA-HONs for ocular application 12
2.7.1. Cell culture and maintenance 12
2.7.2. Cell viability and proliferation assays 12
2.8. Assessment of anti-inflammatory potential of CsA-HON 13
2.8.1. Culture of THP-1 cells and macrophages 14
2.8.2. Immunocytochemistry staining assay 14
2.9. Assessment of anti-angiogenic potential of CsA-HON 15
2.9.1. Cell culture and maintenance 15
2.9.2. Cell viability and proliferation assays 15
2.9.3. Capillary-like tube formation assay 16
2.10. Statistical analysis 16
3. Results and Discussion 17
3.1. Synthesis and characterization of HA-CYS 17
3.2. Synthesis and characterization of HOCs 26
3.3. Characterization of HONs and CsA-HONs 31
3.4. In vitro drug release 38
3.5. In vitro drug permeation 40
3.6. Cytocompatibility evaluation of CsA-HONs for ocular use 42
3.7. Evaluation of the anti-inflammatory activity of CsA-HON1 48
3.8. Assessment of the antiangiogenic potential of CsA-HON1 52
3.9. Commentary for the significance of study 56
4. Conclusions 57
Chapter II. Redox-sensitive oleic acid-hyaluronic acid nanoparticles induce overexpression of lipid accumulation as a synergistic effect to boost the anticancer activity of doxorubicin 58
1. Introduction 58
2. Materials and Methods 60
2.1. Materials 61
2.2. Preparation of DOX-loaded HON 61
2.3. DLS measurement 62
2.4. FE-TEM analysis 62
2.5. In vitro drug release study 63
2.6. Cell culture and maintenance 63
2.7. Oil red O staining and neutral lipid quantification 63
2.8. Neutral lipid droplets staining using BODIPY 493/503 64
2.9. In vitro cytotoxicity 64
2.10. Statistical analysis 65
3. Results and Discussion 66
3.1. Characterizations of GSH-triggered disassembly of HON 66
3.2. Characterizations of DOX-HONs 68
3.3. In vitro drug release study 70
3.4. OA-induced lipid accumulation caused cell death 73
3.5. HOC enhanced the cancer targeting of OA 76
3.6. In vitro cytotoxicity of DOX-loaded formulations 80
3.7. Commentary for the significance of study 83
4. Conclusions 84
References 85