脫層黏土之抑菌功能與生物界面作用機制探討

Jiun-Chiou Wei; 魏郡萩

請用此 Handle URI 來引用此文件： http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63384

完整後設資料紀錄

DC 欄位	值	語言
dc.contributor.advisor	林江珍(Jiang-Jen Lin)
dc.contributor.author	Jiun-Chiou Wei	en
dc.contributor.author	魏郡萩	zh_TW
dc.date.accessioned	2021-06-16T16:38:23Z	-
dc.date.available	2017-10-01
dc.date.copyright	2012-10-01
dc.date.issued	2012
dc.date.submitted	2012-09-27
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dc.identifier.uri	http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63384	-
dc.description.abstract	奈米技術發展數十年，各式各樣的奈米材料已被發現與製備包括天然的無機矽酸鹽層黏土。為了延伸應用無機矽酸鹽層黏土，將其用於蛋白質固定化、抗菌劑與奈米銀粒子/矽酸鹽層黏土複合材之探討。首先，發現牛血清蛋白 (BSA) 與矽酸鹽層黏土可經由直接插層法或逐步插層法製得。逐步插層法為藉由離子交換反應將聚醚胺 (poly-amine) 插層撐開層距隨後可允許蛋白質進入層間。為克服層狀黏土之立體障礙，更進一步發展脫層型奈米矽片用以直接吸附蛋白質，經由逐步程序使生物性巨分子進入層間或以脫層型奈米矽片直接吸附固定於表面，已被成功製備。延伸脫層型奈米矽片之研究，發現奈米矽片具有抵抗多重種類之微生物，包括耐甲氧西林金黃色葡萄球菌及耐銀離子大腸桿菌，由SEM證實奈米矽片會經由物理捕抓方式貼附於細菌表面，更以電子順磁共振光譜證實奈米矽片經光激發可產生1O2進而達到殺菌效果。另外，利用體外彗星測試 (In vitro Comet assay) 評估奈米矽片的基因毒性得知，濃度為1000 μg/mL之奈米矽片與中國倉鼠卵巢癌細胞培養24小時後不會造成DNA損害。利用MTT assay評估奈米矽片的細胞毒性，濃度為1000 μg/mL之奈米矽片和中國倉鼠卵巢癌細胞培養12小時後，顯示為低毒性。更進一步設計新型奈米複合材料-奈米矽片銀，藉由控制奈米矽片與銀之比例，獲得不同尺寸3.6‒35 nm之奈米銀粒子 (AgNP)。將不同比例之奈米矽片銀水溶液進行抗菌測試，發現抗菌能力受銀粒子大小及奈米矽片含量影響。且奈米矽片使銀粒子固定在表面而可能不經由穿透細胞即造成死亡。	zh_TW
dc.description.abstract	Nanotechnology has been developed for decades and various nanomaterials are synthesized including the materials derived from the natural inorganic silicate clays. To extend the applications of nanomaterials, the inorganic silicate clays were used in protein immobilize, antimicrobial agent and silver nanoparticle/silicate clay nanohybrids. In the first part of this study, the hybrids of the model bovine serum albumin (BSA) protein and layered clay were prepared from the direct and stepwise intercalation. The stepwise intercalation involves the uses of the polyamine salts to expand silicate space and subsequently embedding BSA into layered clays. To overcome the steric hindrance of the narrow spacing in the layered clays, the randomized nanoscale silicate platelets (NSP) was used to direct the adsorption of proteins on the surface. Both techniques of embedding large molecules into the silicate gallery and absorbing onto the exposed NSP surface have been successfully developed. We further observed the unexpected antimicrobial behavior for NSP, prepared from the exfoliation of natural clays. A broad spectrum of microorganisms including methicillin-resistant Staphylococcus aureus (MRSA) and silver ion resistance Escherichia coli (E. coli) are tested. The NSP enabling of physically adhering onto to microbe surfaces and resulting antimicrobial ability is attributed to the singlet oxygen (1O2) generation tested for the photoexcited NSP using spectroscopic methods (EPR spin-trapping). The material safety on toxicity was investigated by using the Comet assay test in vitro and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The Comet assay showed no DNA damage after incubation with 1000 μg/mL of NSP and MTT assay showed a low cytotoxicity below 1000 μg/mL and a dose-dependent effect. Further manipulating the function of silicate clays, we found that the silver nanoparticles (AgNP) could be tailored by using NSP as the supports. The amount of adding the thin silicate platelets at the composition ratio from 0.5/99.5 to 50/50 of Ag/silicate can largely affect the formation of the AgNP size ranging from 3.6 nm to 35 nm. The evaluation for antibacterial activity has shown a silver size-dependent effect on controlling the growth of dermal pathogens. The antimicrobial efficacy can be corresponded to the size of AgNP and the amount of NSP presence.	en
dc.description.provenance	Made available in DSpace on 2021-06-16T16:38:23Z (GMT). No. of bitstreams: 1 ntu-101-D95549012-1.pdf: 6483265 bytes, checksum: 7fe2f909e5fba212371bc64fce4116bc (MD5) Previous issue date: 2012	en
dc.description.tableofcontents	CONTENT I LIST OF TABLES III LIST OF SCHEMES IV LIST OF FIGURES V 摘要 IX ABSTRACT XI CHAPTER 1. INTRODUCTION AND LITERATURE REVIEW 1 1.1. INTRODUCTION OF NANOMATERIAL 1 1.2. IMMOBILIZING BIOMATERIAL 3 1.2.1. Organically Modified Layered Silicate (OLS) 4 1.2.2. Bio-polymers Immobilization in Layere Silicate 6 1.2.2.1. Immobilization by Chemical Methods 7 1.2.2.2. Immobilization by Physical Methods 7 1.2.2.3. Smectitic Clay as Substrate for Bio-polymers Immobilization 8 1.3. FUNCTIONALITY OF NANOMATERIALS 11 1.3.1. Antibacterial Property 12 1.3.1.1. Nano-antimicrobial Materials of Spherical Shape 14 1.3.1.2. Nano-antimicrobial Materials of Rod/Fiber Shape 15 1.3.1.3. Nano-antimicrobial Materials of Lamellar Shape 16 1.3.2. Cytotoxicity Property 19 1.3.2.1.Cytotoxicity of Nanoparticles 20 1.3.2.2. Cytotoxicity of Layer Materials‒smectite Clays 21 1.4. SYNTHESIS OF SILVER NANOPARTICLES 22 1.4.1. Synthesis of AgNP 23 1.4.2. Silver as Antibacterial Material 25 1.4.2.1. Bactericidal Mechanism of Silver 25 1.4.2.2. The Size and Shape Effect on the Antibacterial activity of AgNP 26 CHAPTER 2. EXPERIMENTAL SECTION 28 2.1. MATERIALS 28 2.2. PREPRATION OF PROTEIN-CLAY HYBRIDS 32 2.3. ANTIBACTERIAL ACTIVITY OF NSP 33 2.4. ROS DETECTED 34 2.5. TOXICITY OF NSP 35 2.6. PREPARATION OF Ag/CLAY HYBRID 37 2.7. PREPARATION OF Ag/POLYMER HYBRID 37 2.8. TEM EXAMINATION 38 2.9. INSTRUMENTS AND ANALYSES 38 2.10. STATISTICAL ANAYLSIS 39 CHAPTER 3. RESULTS AND DISCUSSION 40 3.1. NOVEL PREPARATION OF PROTEIN-CLAY HYBRIDS 40 3.1.1. pH-dependence in the Process of BSA Intercalation 41 3.1.2. Direct Intercalation of BSA with Pristine Caly 42 3.1.3. Two-Step Intercalation with the POP-Modified Clay with Property of Inverse Dispersion Temperature in Water 43 3.1.4. Second Step of BSA Replacement to a Compressed or Uncompressed Conformation 45 3.1.5. Direct Observation of BSA/MMT Hybrid by AFM 49 3.1.6. Direct Absorption of BSA with Exfoliated Silicates 50 3.1.7. Conclusion 54 3.2. INHIBITION OF BACTERIAL GROWTH BY THE EXFOLIATED CLAYS AND OBSERVATION OF PHYSICAL CAPTURING MECHANISM 55 3.2.1. Material Description: Preparation and Purification of NSP 56 3.2.2. Antimicrobial Potency 60 3.2.3. Physical Trapping Mechanism 64 3.2.4. Observation of Singlet Oxygen Species by the Exfoliated Clay Silicate Platelets 67 3.2.5. Evaluation on Cytotoxicity and Genotoxicity of the NSP 71 3.2.6. Conclusion 73 3.3. SIZE-CONTROLLED SYNTHESIS AgNP ON CLAY FOR BIOMEDICAL APPLICATIONS 75 3.3.1. Synthesis of AgNP/nanoplatelet Nanohybrids 76 3.3.2. Growth Inhibition Effect Against Bacteria 78 3.3.3. SEM and TEM Morphology 80 3.3.4. Conclusion 86 CHAPTER 4. CONCLUSIONS 87 FUTURE WORK 89 CURRICULUM VITAE 90 REFERENCES 94
dc.language.iso	en
dc.title	脫層黏土之抑菌功能與生物界面作用機制探討	zh_TW
dc.title	Mechanistic Studies on Antimicrobial Property and Bio-Interaction of Exfoliated Nanoscale Silicate Platelets	en
dc.type	Thesis
dc.date.schoolyear	101-1
dc.description.degree	博士
dc.contributor.oralexamcommittee	郭炳林(Ping-Lin Kuo),徐善慧(Shan-hui Hsu),鄭如忠(Ru-Jong Jeng),謝國煌(Kuo-Huang Hsieh),彭福佐(Fu-Chuo Peng)
dc.subject.keyword	抗菌,奈米矽片,活性氧化物,電子順磁共振光譜,細胞毒性,基因毒性,銀奈米粒子,	zh_TW
dc.subject.keyword	Antimicrobial,nanosilicate platelets,Reactive oxygen species,Electron paramagnetic resonance,cytotoxicity,genotoxicity,silver nanoparticles,	en
dc.relation.page	99
dc.rights.note	有償授權
dc.date.accepted	2012-09-27
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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