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In Vitro and In Vivo Toxicity Mechanisms of ZnO and TiO2 Nanoparticles : 산화아연 및 이산화티타늄 나노입자의 독성학적 기전연구
DC Field | Value | Language |
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dc.contributor.advisor | 조명행 | - |
dc.contributor.author | Kyeong-Nam Yu | - |
dc.date.accessioned | 2017-07-13T16:44:29Z | - |
dc.date.available | 2017-07-13T16:44:29Z | - |
dc.date.issued | 2015-08 | - |
dc.identifier.other | 000000067013 | - |
dc.identifier.uri | https://hdl.handle.net/10371/120226 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 수의학과, 2015. 8. 조명행. | - |
dc.description.abstract | With the growth of nanotechnology, there has been a tremendous development in the diverse applications of nanomaterials such as antibacterial materials, drug delivery systems, electronics, cosmetics, etc. Nanomaterials have dramatically different physicochemical properties compared to fine particles of the similar composition. The smaller size of nanomaterials ensures that a large portion of atoms will be on the particle surface. Since surface properties, such as electronic structure, energy level and reactivity are quite different from interior states, the bioactivity of nanoparticles may likely differ from that of the fine size analogue. The nanomaterials is categorized as polymer based, ceramics, oxide or metal materials, carbon-based and silicon. However, the toxicity studies of nanomaterials have not fully elucidated. In this research, we studied the toxicity mechanisms of ZnO and TiO2 nanoparticles.
In Part I, zinc oxide (ZnO) nanoparticles, which have been used in an increasing number of industrial products such as paint, coating and cosmetics, and in other biological applications, were chosen for toxicity mechanism. There have been many suggestions of ZnO nanoparticles toxicity paradigm but the underlying molecular mechanisms of the toxicity of ZnO nanoparticles remain unclear. This study was done to determine the potential toxicity of ZnO nanoparticles and to assess the toxicity mechanism in normal skin cells. Synthesized ZnO nanoparticles generated reactive oxygen species (ROS), as determined by electron spin resonance. After uptake into cells, ZnO nanoparticles induced ROS in a concentration- and time-dependent manner. To demonstrate ZnO nanoparticles toxicity mechanism related to ROS, we detected abnormal autophagic vacuoles accumulation and mitochondria dysfunction after ZnO nanoparticles treatment. Furthermore mitochondria membrane potential and adenosine triphosphate (ATP) production were decreased by the treatment of ZnO nanoparticles. Our results suggested that ZnO nanoparticles led to cell death through autophagic vacuole accumulation and mitochondria damage in normal skin cells via ROS induction. Nanomaterials are also used in diverse fields including food, cosmetic, and medical industries. Titanium dioxide (TiO2) nanoparticles are widely used, but their effects on biological systems and mechanism of toxicity have not been elucidated fully. In Part II, we report the toxicological mechanism of TiO2 nanoparticles in cell organelles. Human bronchial epithelial cells (16HBE14o-) were exposed to 50 and 100 μg/mL TiO2 nanoparticles for 24 and 48 h. Our results showed that TiO2 nanoparticles induced endoplasmic reticulum (ER) stress in the cells and disrupted the mitochondria-associated endoplasmic reticulum membranes (MAMs) and calcium ion balance, thereby increasing autophagy. In contrast, an inhibitor of ER stress, tauroursodeoxycholic acid (TUDCA), mitigated the cellular toxic response, suggesting that TiO2 nanoparticles promoted toxicity via ER stress. This novel mechanism of TiO2 nanoparticles toxicity in human bronchial epithelial cells suggests that further exhaustive research on the harmful effects of these nanoparticles in relevant organisms is needed for their safe application. In the workplace, the primary exposure route for TiO2 nanoparticles is inhalation through the respiratory system. Because TiO2 nanoparticles have different physiological properties, in terms of size and bioactivity, their toxic effects in the respiratory system must be determined. In Part III, to determine the toxic effect of inhaled TiO2 nanoparticles in the lung and the underlying mechanism, we used a whole-body chamber inhalation system to expose A/J mice to TiO2 nanoparticles for 28 days. During the experiments, the inhaled TiO2 nanoparticles were characterized using a cascade impactor and transmission electron microscopy. After inhalation of the TiO2 nanoparticles, hyperplasia and inflammation were observed in a TiO2 nanoparticles treated murine lung. To determine the biological mechanism of the toxic response in the lung, we examined endoplasmic reticulum (ER) and mitochondria in lung tissues. The ER and mitochondria were disrupted and dysfunctional in the TiO2 nanoparticles exposed lung leading to abnormal autophagy. In summary, we assessed the potential risk of TiO2 nanoparticles in the respiratory system, which contributed to our understanding of the mechanism underlining TiO2 nanoparticles toxicity in the lung. | - |
dc.description.tableofcontents | Contents
Abstract i Contents iv List of Figures vi List of Tables viii General Introduction 1 Reference 7 I. Zinc oxide nanoparticle induced autophagic cell death and mitochondrial damage via reactive oxygen species generation 13 Abstract 14 Introduction 15 Materials and Methods 17 Results 22 Discussion 38 Supplementary Figures 41 Reference 45 II. Titanium dioxide nanoparticles induce endoplasmic reticulum stress-mediated autophagic cell death via mitochondria-associated endoplasmic reticulum membrane disruption in normal lung cells 51 Abstract 52 Introduction 53 Materials and Methods 55 Results 61 Discussion 81 Supplementary Figures 85 Reference 91 III. Inhalation of titanium dioxide nanoparticles induces endoplasmic reticulum stress-mediated autophagy and inflammation in mice 98 Abstract 99 Introduction 100 Materials and Methods 102 Results 107 Discussion 132 Reference 135 국문 초록 142 Publication 145 List of Figures I. Zinc oxide nanoparticle induced autophagic cell death and mitochondrial damage via reactive oxygen species generation 13 Figure 1. Characterization of the synthesized ZnO nanomaterial 23 Figure 2. ESR, CLSM and FACS data for detect of ROS generation measurement 26 Figure 3. Confirmation of the induction of autophagy in the presence of ZnO 29 Figure 4. ptfLC3 transfection data for confirmation of autophagy 32 Figure 5. Effect of ROS on mitochondria 35 Figure S1. ESR data for detection of singlet oxygen 41 Figure S2. bio-TEM images for non-vacuole with NAC chemicals 42 Figure S3. CLSM data for confirmation of autophagy after transfection of ptfLC3 43 Figure S4. Monitoring of mitochondria activity 44 II. Titanium dioxide nanoparticles induce endoplasmic reticulum stress-mediated autophagic cell death via mitochondria-associated endoplasmic reticulum membrane disruption in normal lung cells 51 Figure 1. Characterization of Titanium Dioxide nanoparticles and detection of reactive oxygen species (ROS) generation in Human Bronchial EpithelialCells 62 Figure 2. Titanium Dioxide nanoparticles treatment Elevated Endoplasmic Reticulum (ER) stress in Human Bronchial Epithelial Cells 66 Figure 3. Titanium dioxide nanoparticles damaged the connection between the endoplasmic reticulum (ER) and mitochondria 70 Figure 4. Titanium dioxide nanoaprticles Induced autophagy in human bronchial epithelial Cells 75 Figure 5. Treatment with the endoplasmic reticulum (ER) stress Inhibitor (TUDCA) affects cell biogenesis 79 Figure 6. Schematic diagram of titanium dioxide nanoparticles (TiO2-NP) toxicity in normal lung cells 84 Figure S1. X-ray diffraction XRD data of TiO2-NP 85 Figure S2. Intracellular ROS detection and cell viability monitoring 86 Figure S3. TiO2-NP treatment effect to ER condition 88 Figure S4. TiO2-NP affect the mitochondria-associated endoplasmic retuculum (ER) membranes (MAMs) 89 Figure S5. ER condition detected after ER stress inhibition in TiO2 treated cells 90 III. Inhalation of titanium dioxide nanoparticles induces endoplasmic reticulum stress mediated autophagy and inflammation in mice 98 Figure 1. TiO2 nanoparticles inhalation study 108 Figure 2. Inhaled TiO2 nanoparticles induced pulmonary inflammation 116 Figure 3. Inhaled TiO2 nanoparticles induced cell organelle disruption and ER stress in lungs 124 Figure 4. Inhaled TiO2 nanoparticles activated autophagy in lungs 129 List of Tables II. Titanium Dioxide Induces Endoplasmic Reticulum Stress-Mediated Autophagic Cell Death via Mitochondria-Associated Endoplasmic Reticulum Membrane Disruption in Normal Lung Cells Table 1. Physical characterization of Titanium dioxide nanoparticles 64 III. Inhalation of titanium dioxide induces endoplasmic reticulum stress mediated autophagy and inflammation in mice Table 1. TiO2 nanoparticles concentration of animal exposure chamber 112 Table 2. Monitoring of TiO2 nanoparticles size in inhalation experiment 113 Table 3. Serum biochemical analysis after TiO2 nanoparticles inhalation 120 Table 4. Blood analysis after TiO2 nanoparticles inhalation 121 | - |
dc.format | application/pdf | - |
dc.format.extent | 3057366 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | Nanomaterials | - |
dc.subject | Mechanism of nano-toxicity | - |
dc.subject | ZnO nanoparticles | - |
dc.subject | TiO2 nanoparticles | - |
dc.subject | cell death | - |
dc.subject.ddc | 636 | - |
dc.title | In Vitro and In Vivo Toxicity Mechanisms of ZnO and TiO2 Nanoparticles | - |
dc.title.alternative | 산화아연 및 이산화티타늄 나노입자의 독성학적 기전연구 | - |
dc.type | Thesis | - |
dc.contributor.AlternativeAuthor | 유 경남 | - |
dc.description.degree | Doctor | - |
dc.citation.pages | viii, 152 | - |
dc.contributor.affiliation | 수의과대학 수의학과 | - |
dc.date.awarded | 2015-08 | - |
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