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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56601
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dc.contributor.advisor林江珍(Jiang-Jen Lin)
dc.contributor.authorYu-Hsuan Leeen
dc.contributor.author李宇軒zh_TW
dc.date.accessioned2021-06-16T05:37:08Z-
dc.date.available2017-08-17
dc.date.copyright2014-08-17
dc.date.issued2014
dc.date.submitted2014-08-12
dc.identifier.citationReferences
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56601-
dc.description.abstract近年來奈米銀粒子(AgNP)已被利用於許多抗菌產品上,例如紡織物、淨水器、體內骨釘等等。許多研究發現AgNP具有良好的抗菌性,但其高活性也會對人體造成傷害。本實驗室開發片狀矽酸鹽黏土(NSP)使奈米銀粒子(AgNP)穩定在NSP表面上,合成出奈米混成材料-奈米矽片銀粒子(AgNP/NSP)。本研究對AgNP/NSP進行一系列毒性相關研究,利用人永生化表皮细胞(HaCaT cells)鑑定其細胞毒性。以MTT assay測試評估不同的AgNP/NSP重量比的細胞毒性,其結果顯示NSP的重量百分比越高,會造成較高的細胞毒性。但根據我們先前的研究,高百分比的NSP,合成出的AgNP/NSP其AgNP粒子較小且抗菌能力較好。綜合分析其對於各種不同格蘭氏陰性與格蘭氏陽性菌的最低抑菌濃度,AgNP/NSP重量百分比7/93在其最低抑菌濃度(AgNP: 3.9 ppm)有最高的HaCaT cells細胞存活率(95%)。
此外,利用四種不同之測試系統對AgNP/NSP的基因毒性做評估,分別為: 體外哺乳類細胞的染色體異常分析法 (In vitro Chromosome aberration test) ,體外彗星測試 (In vitro Comet assay) ,體外微生物基因突變分析 (In vitro Ames test) 和體內微核試驗 (In vivo micronucleus assay) 。由以上基因毒性測試下,AgNP/NSP皆無觀察到染色體異常與DNA的損害。這些結果印證AgNP/NSP為安全奈米混合材料可發展於生醫應用上。由TEM的觀察,我們發現AgNP/NSP低毒性機制來自於NSP的高表面積,其高表面積可阻止AgNP穿透細胞膜,並降低基因毒性。綜合以上各點,AgNP/NSP是安全且有效的抗菌劑適合發展生醫應用。
zh_TW
dc.description.abstractIn recent years, silver nanoparticles (AgNP) has been used in many antibacterial products, such as drinking water purification and bone cement or treatment of many skin conditions and the use as antibacterial textile. Since nanomaterials and nanotechnologies are expected to yield numerous health and health care advances, tighter the regulation of nanotechnology has been demanded.
  Silver nanoparticles (AgNP) are known for excellent bactericidal properties, but there are the concerns of the possible adversity toward normal cells. We have developed the one-nanometer-thick silicate platelets (NSP) from the natural silicate clays and employed them as the high surface support for enhancing antimicrobial efficacy but mitigating the toxicity of normal cells. The NSP nanoplatelets were in the geometric dimension of ca. 80 × 80 × 1 nm3 and possessing of intensive ionic charges on the local surface. The synthesis of the nanohybrids of AgNPs immobilized on NSP (AgNP/NSP) were proven to have the enhanced antimicrobial properties against the growth of bacteria including Gram-positive and Gram-negative. In this study, we synthesized AgNP/NSP nanohybrids in three different weight ratios (1/99, 7/93 and 15/85) and particle size in the range of 3.8-9.3 nm in diameter. Cytotoxicity by using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and genotoxicity by the methods of chromosomal aberration, comet, Ames and micronucleus assays were determined. Overall, AgNP/NSP showed none of significant cytotoxicity nor DNA damage at the bactericidal dose. By comparison, the synthesized AgNP in polymeric organic dispersants were shown high cytotoxicity and genotoxicity by the same tests. Due to the high surface support of the NSP, NSP prevents the immobilized AgNP enter cell membrane, which lower the risk associated in AgNP toxicity. Hence, the newly developed AgNP/NSP nanohybrids demonstrated low toxicity based on the cytotoxic and genotoxic tests and suitability for potential uses in biomedical areas.
en
dc.description.provenanceMade available in DSpace on 2021-06-16T05:37:08Z (GMT). No. of bitstreams: 1
ntu-103-R01549001-1.pdf: 5576000 bytes, checksum: 4f1f1586de63b8d310e86b58170f9320 (MD5)
Previous issue date: 2014
en
dc.description.tableofcontents口試委員會審定書
Acknowledgements
中文摘要
Abstract
Contents
Figure captions
Table captions
Scheme captions
Chapter 1 Introduction and Literature Review 1
1.1. Introduction of Nanomaterials 1
1.2. Introduction of Antibacterial Nanomaterials 2
1.3. Introduction of Silver Nanoparticles 4
1.3.1. Antibacterial Mechanism of Silver Nanoparticles 5
1.3.2. Toxicity of Silver Nanoparticles 7
1.4. Introduction of Nanoscale Silicate Platelets 8
1.5. Synthesis of AgNP/NSP Nanohybrids 9
1.6. Antimicrobial Ability of AgNP with Various Sizes 11
1.7. Cytotoxicity 13
1.7.1. MTT Assay 13
1.7.2. Cytotoxicity of Metal Nanoparticles 14
1.8. Genotoxicity 14
1.8.1. Ames Mutagenicity Test 16
1.8.2. Micronucleus Assay 18
1.8.3. Comet Assay 18
1.8.4. Chromosome Aberration Test 19
1.9. Cytotoxicity and Genotoxicity of NSP 19
1.10. Research Objectives 20
Chapter 2 Experimental Section 21
2.1. Materials 21
2.2. Synthesis of AgNP/NSP Nanohybrids 21
2.3. Synthesis of AgNP/SMA Nanohybrids 22
2.4. Transmission Electron Microscopy 23
2.5. Sources of Bacteria 23
2.6. Evaluation of Antibacterial Activity 23
2.7. Evaluation of Bactericidal Ability for AgNP with Various Sizes 24
2.8. Process of Preparing TEM Samples for Bactericidal Mechanism of AgNP 25
2.9. Cell Culture and Preparation 25
2.10.Cytotoxicity Assay 26
2.10.1. MTT Viability Assay 26
2.11. Genotoxicity assay 26
2.11.1. Ames test in vitro 26
2.11.2. Chromosome Aberration Test in vitro 27
2.11.3. Comet assay in vitro 28
2.11.4. Micronucleus assay in vivo 29
Chapter 3 Results and Discussion 30
3.1. Cytotoxicity Assay of AgNP/NSP in vitro 30
3.2. Genotoxicity Assay of AgNP/NSP 32
3.2.1. Chromosome Aberration test of AgNP/NSP in vitro 32
3.2.2. Comet Assay of AgNP/NSP in vitro 34
3.2.3. Micronucleus Assay of AgNP/NSP in vivo 36
3.2.3. Ames test of AgNP/NSP in vitro 38
3.3. Investigation for the Possible Bactericidal Mechanism of AgNP 47
Chapter 4 Conclusion 53
Chapter 5 References 56
dc.language.isoen
dc.subject低基因毒性zh_TW
dc.subject奈米銀粒子zh_TW
dc.subject奈米混合材料zh_TW
dc.subject奈米矽片zh_TW
dc.subject抗菌zh_TW
dc.subject低細胞毒性zh_TW
dc.subjectsilicate nanoplateletsen
dc.subjectnanohybridsen
dc.subjectlow genotoxicityen
dc.subjectlow cytotoxicityen
dc.subjectsilver nanoparticlesen
dc.subjectantimicrobialen
dc.title銀粒子/矽片奈米複合物之細胞與基因毒性與機制探討zh_TW
dc.titleStudies on Cytotoxicity and Genotoxicity of Silver Particle/Silicate Platelet Nanohybridsen
dc.typeThesis
dc.date.schoolyear102-2
dc.description.degree碩士
dc.contributor.oralexamcommittee謝國煌(Kuo-Huang Hsieh),徐善慧(Shan-hui Hsu),彭福佐(Fu-Chuo Peng)
dc.subject.keyword奈米混合材料,奈米銀粒子,奈米矽片,低細胞毒性,低基因毒性,抗菌,zh_TW
dc.subject.keywordnanohybrids,silver nanoparticles,silicate nanoplatelets,antimicrobial,low cytotoxicity,low genotoxicity,en
dc.relation.page60
dc.rights.note有償授權
dc.date.accepted2014-08-12
dc.contributor.author-college工學院zh_TW
dc.contributor.author-dept高分子科學與工程學研究所zh_TW
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