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標題: 奈米銀脫層黏土之抗菌機制及應用
Antimicrobial mechanism and applications of silver nanoparticles on the exfoliated platelet clay
作者: 包翊絹
Bau, I-Juian
關鍵字: 奈米銀;silver nanoparticles;抗菌機制;掃描式電子顯微鏡;金黃色葡萄球菌;大腸桿菌;沙門氏桿菌;鮑氏不動桿菌;antimicrobial mechanism;Scanning Electron Microscope;Staphylococcus aureus;Escherichia coli;Salmonella typhimurium;Acinetobacter baumannii
出版社: 生命科學系所
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銀或含銀化合物能抗菌力,而經奈米化的銀粒子也具有殺菌的功效,但其抗菌機制目前尚不清楚。本研究使用的奈米銀黏土材料AgNP/NSP與AgNP/SWN是使用NSP脫層黏土及SWN蒙脫土作為分散劑,以防止奈米銀粒子在製程中產生聚集,藉以研究奈米銀的抗菌機制。首先利用AgNP/NSP與AgNP/SWN對具多重抗藥性菌株鮑氏不動桿菌(Acinetobacter baumannii)及具有銀離子抗性之大腸桿菌菌株( E. coli strain J53 pMG101)作抑菌能力分析並與硝酸銀的抗菌能力作比較,比較後的結果顯示在相同銀濃度下, AgNP/NSP對此兩種菌均具有最好的抑菌能力。以掃瞄式電子顯微鏡(Scanning Electron Microscope)觀察經奈米銀黏土處理的細菌發現材料將其表面包覆,且使細菌聚集。加入propidium iodide (PI)後24小時,吸附於奈米銀黏土上的細菌內的DNA並無呈現紅色螢光,證明細菌的細胞膜大致完好,仍可抵抗PI進入細菌內,但是否因細胞膜仍有輕微損害而導致抑制細菌生長仍不清楚。使用大腸桿菌E. coli strain ML-35p,觀察到奈米銀黏土的確會增加細菌細胞膜的通透性,因此推測奈米銀黏土材料藉由此方式影響細菌的分裂而達到殺菌的功效。也發現只有高於0.1wt%奈米銀黏土可顯著降低β-galactosidase的酵素活性,證明奈米銀導致蛋白質變性的能力並不強。除細菌外,也發現AgNP/NSP與AgNP/SWN具有抑制真菌生長之特性並會將真菌表面包覆。另外在奈米銀脫層黏土材料應用於治療家禽細菌性下痢以取代抗生素的實驗中,顯示當雛雞感染沙門氏桿菌後的十八小時內,經口投與AgNP/NSP,可能可以減緩細菌經血液蔓延至其他器官而達到治療的效果。使用轉移子 ( transposon ) 對奈米銀粒子抗性基因進行篩選,也未發現突變細菌株,此結果顯示奈米銀並無基因毒性。最後在奈米銀黏土材料的致突變性試驗 (Ames test )中,結果顯示在0.5mg/plate 劑量以下不具有致突變的作用。而未來將更深入探討奈米銀黏土的抑菌機制並將其應用於細菌感染的治療上。

Silver or silver compound can inhibit bacteria growth. Silver nanoparticle also has the bactericidal effect but the antibacterial mechanism is still unknown. AgNP/NSP and AgNP/SWN, the materials used in this article, are synthesized by using nano silicate platelet clays and artificial organic clays as dispersing to stabilize and immobilize silver nanoparticles and consequently used to study the antibacterial mechanism of silver nanoparticles. Firstly, the growth inhibition capability of AgNP/NSP and AgNP/SWN to multidrug-resistant Acinetobacter baumannii and silver ion-resistant E. coli strain J53 pMG101 were analyzed and compared with silver nitrate. The result showed that among these three silver compound AgNP/NSP has the best bactericidal effect under the same silver concentration.Observation by using Scanning Electron Microscope(SEM) revealed that AgNP/NSP encapsulated the bacteria and form multiple clay-cells aggregates. Absorbed bacteria on AgNP/NSP repelled the entry of propiodium iodide(PI) indicating that bacteria cell membrane was not ruptured. But whether the mild lesion of cell membrane leads the growth retardation is still unknown. By using E. coli strain ML-35p, we found that Ag nanoparticles clay indeed could increase the membrane permeability.Therefore we speculate that Ag nanoparticles clay has bactericidal effect through interfering intracellular homeostasis. We also find only higher than 0.1wt% Ag nanoparticles clay can obviously decrease β-galactosidase activity and this result demonstrated the oxidization of functional proteins by Ag nanoparticles was not strong. Besides bacteria , AgNP/NSP and AgNP/SWN also could inhibit the growth of infectious fungi, including Trichophyton rubrum、Trichophyton fluviomuniense、Trichophyton mentagrophytes、Microsporum canis and Microsporum gypseum , the common pathogens of Tinea pedis. For the substitution of antibiotics, Ag nanoparticles clay was applied to cure fowl diarrhea after infection with salmonella. After eighteen hours, AgNP/NSP via oral route retarded bacteria spread to other organ. The Ag nanoparticles- resistant bacteria was not found, showing the safety of clinical application. We further demonstrated that geno-safety of AgNP/NSP by Ames test, showing lower than 0.5mg/plate dose have no genetic mutation. In the future , we will further explore the mechanism of bactericidal effect of AgNP/NSP and apply Ag nanoparticles clay to clinical treatment of bacterial infection.
其他識別: U0005-1908200812315100
Appears in Collections:生命科學系所

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