Efficacy and toxicity of drug-loaded nanoparticle using hepato-mimetic microfluidic chip system
- 주제(키워드) biomimetic microfluidic system , cell viability , ROS sensor , anti-cancer efficacy , hepatotoxicity , nanoparticles , IC50 value , doxorubicin , paclitaxel
- 주제(DDC) 615.1
- 발행기관 아주대학교
- 지도교수 이범진
- 발행년도 2022
- 학위수여년월 2022. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000031648
- 본문언어 한국어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The aim of this study is to evaluate the anticancer efficacy of doxorubicin (DOX) and the toxicity of paclitaxel (PTX) according to formulation and cell culture conditions using a biomimetic microfluidic chip system. Using a ROS sensor chip-based biomimetic microfluidic system, HepG2 liver cancer cells were treated with drug-loaded human serum albumin-oleic acid nanoparticles (AONs) in serum-free medium at 37 °C, 5% CO for 24 h. The HepG2 seeded chip was coupled to the microfluidic system. Cell uptake and viability were assessed for free drug (DOX, PTX) and doxorubicin loaded nanoparticles (DOX-AON), paclitaxel loaded nanoparticles (PTX-AON) under static or dynamic conditions (5 dyne/cm2). Intracellular drug uptake was observed using a confocal laser scanning microscope, and cell viability according to drug concentration was evaluated by MTT assay. In addition, the ROS generated in cells treated with free drugs (DOX, PTX) under dynamic conditions was confirmed in a dose-dependent manner using the ROS sensor, and the anticancer efficacy and hepatotoxicity of the drug were evaluated in relation to the cell viability. The efficacy and toxicity of drug-loaded nanoparticles varied in a dose-dependent manner in HepG2 according to the degree of ROS occurrence. Drug-loaded nanoparticles increased intracellular fluorescence intensity and decreased cell viability compared to free drug under dynamic conditions. The IC50 values of DOX were 13.81 ug/mL, 9.77 ug/mL, and 6.026 ug/mL in static, dynamic, and DOX-AONs-treated conditions, respectively. The IC50 values of PTX were 50.53 ug/mL, 38.35 ug/mL, and 20.67 ug/mL in the same order as DOX (static, dynamic, AONs). The concordance rates of cell viability and ROS were 78.95% for DOX and 92.61% for PTX. The sensor chip-based biomimetic microfluidic system was useful to evaluate the real-time efficacy and toxicity of free drugs and drug-loaded AONs on human tissues.
more목차
1. Introduction 1
2. Materials and Methods 4
2.1. Materials 4
2.2. Preparation of AONs, DOX-AONs, and PTX- AONs 4
2.2.1. Synthesis of AOC 4
2.2.2. Converting AOC to AONs 7
2.2.3. Preparation of DOX-AONs and PTX-AONs 7
2.3. Characterization of AONs, DOX-AONs, and PTX-AONs 8
2.3.1. FT-IR 8
2.3.2. Particle size and zeta potential 9
2.3.3. Drug loading content (DC) and encapsulation efficiency (EE) 9
2.3.4. Morphology of AONs: FE-TEM, FE-SEM 10
2.4. Cell culture 11
2.5. Biomimetic microfluidic experiment 12
2.5.1. Biomimetic microfluidic system calibration 12
2.5.2. Cell viability assay 18
2.5.3. Sensing of ROS generation 19
2.6. Confocal laser scanning microscopy 19
3. Results and discussions 21
3.1. Physicochemical properties: AONs, DOX-AONs, and PTX-AONs 21
3.1.1. Identification of oleic acid conjugated human serum albumin 21
3.1.2. Particle size, zeta potential and LD, EE analysis 23
3.2. Confirmation of cell viability by the treatment of AONs 30
3.3. Efficacy of hepatocarcinoma by DOX 32
3.4. Hepatotoxicity induced by PTX 39
4. Conclusions 45
5. References 46
국문초록 52
