綠色介質活化含銀非晶相固體對抗藥性致病菌之抑菌研究

Antibacterial activity of silver-containing non-crystalline solids by green-mediated against drug-resistant pathogenic bacterial

李忠霖(Chung-Lin Lee)

指導教授 ： 石啟仁

高雄醫學大學/藥學院/香粧品學系碩士班/碩士(2018年)

摘要

抗藥型致病菌的危害日益嚴重；2014年10月英國抗藥性研究組織以「Review on Antimicrobial Resistance」為題，發布總結報告預測於 2050 年時，每年將會有一千萬人口因抗藥型致病菌感染致死。相較於抗生素，奈米銀(AgNPs)因不具抗藥性，為極具潛力之抗菌候選材料。考量其生物親和性，合成 AgNPs 研究主要以綠色介質合成為主，包含蛋白質合成和植物萃取合成。然而相關研究大多以硝酸銀作為Ag+ 源，製程中易造成凝聚、粗化及析出等現象，導致成分不均。含銀非晶相固體(silver-containing non-crystalline solid, SCNS)兼具骨傳導、誘導和易釋放離子的特性，常應用於硬組織之填補、修復和抗菌；然而受限於銀離子釋放能力不足，導致抗菌效果不佳。
本研究先合成 SCNS，並以 AgNO3比較；分別使用蛋白和檄樹葉萃取活化以形成 AgNPs 試劑，並比較其對抗藥型致病菌的抑菌效能。實驗結果顯示：Ag+ 源和 pH 值影響 AgNPs 的形貌和表面電漿共振；其中以蛋白與檄樹葉萃取活化形成的AgNPs 原始粒徑約為 5 nm，Ag+ 濃度最大可達68 ppm。經 Time-Kill curve (時間抑菌曲線)和Colony-forming capacity assay (菌落形成能力試驗)測定其對於3種致病菌 (金黃色葡萄球菌、綠膿桿菌、克雷伯氏鏈球菌) 與4種抗藥型致病菌(耐甲氧西林金黃色葡萄球菌、對萬古黴素感受性下降之金黃色葡萄球菌)的抑菌結果顯示：以蛋白萃取活化形成AgNPs時，則SCNS抑菌性優於AgNO3；以檄樹葉萃取活化形成AgNPs時，則AgNO3抑菌性優於SCNS。

關鍵字

綠色合成 ； 奈米銀 ； 含銀非晶相固體 ； 抗藥性致病菌

並列摘要

Drug-resistant pathogenic bacteria are becoming more and more dangerous. In October 2014, a summary report titled “Review on Antimicrobial Resistance” was issued by the British Drug Resistance Research Group. It was predicted that 1 billion people will die from drug-resistant pathogenic bacteria infections by 2050. Compared with antibiotics, silver nanoparticles (AgNPs) are potential antibacterial candidates due to their non-resistance. Considering its biological affinity, synthesis of AgNPs is mainly based on green media synthesis, including protein-mediated synthesis and plant-mediated synthesis. However, most of the related researches use silver nitrate as Ag+ precursor, and the process is easy to produce agglomeration, coarsening, and precipitation, resulting in uneven composition. Silver-containing non-crystalline solid (SCNS) having the characteristics of bone conduction, bone induction, and easily ions release is often applied to the filling, repair, and antibacterial of hard tissues. However, due to the insufficient release of silver ions, SCNS has a poor antibacterial efficacy.
In this study, SCNS was first synthesized, and AgNO3 was used as a control group. Then SCNS combining with Casein or Morinda citrifolia leaves were extracted and activated to form AgNPs reagents, and their antibacterial efficacy against drug-resistant pathogenic bacteria were compared. The experimental results show that the Ag+ precursor and pH value affect the morphology and surface plasmon resonance (SPR) of AgNPs. The primary size of AgNPs formed by Casein extraction and Morinda citrifolia leaves extract and activation is about 5 nm, and the maximum Ag+ concentration is up to 68 ppm. The antibacterial efficacy of the AgNPs against three pathogenic bacterial and four types drug-resistant pathogenic bacterial were determined by Time-Kill curve and Colony-forming capacity assay. The experimental results show that when the AgNPs were extracted and activated by Casein, the SCNS was superior to AgNO3 in antibacterial efficacy; when the AgNPs were extracted and activated by Morinda citrifolia leaves, AgNO3 was superior to SCNS

並列關鍵字

Green synthesis ； Silver nanoparticles ； Silver-containing non-crystalline solid ； Drug-resistant pathogenic bacterial

參考文獻

1. Marambio-Jones, Catalina, and Eric MV Hoek. "A review of the antibacterial effects of silver nanomaterials and potential implications for human health and the environment." Journal of Nanoparticle Research 12.5 (2010): 1531-1551.

2. Dallas, Panagiotis, Virender K. Sharma, and Radek Zboril. "Silver polymeric nanocomposites as advanced antimicrobial agents: classification, synthetic paths, applications, and perspectives." Advances in colloid and interface science 166.1-2 (2011): 119-135.

3. Choi, Okkyoung, et al. "The inhibitory effects of silver nanoparticles, silver ions, and silver chloride colloids on microbial growth." Water research 42.12 (2008): 3066-3074.

4. Ahamed, Maqusood, Mohamad S. AlSalhi, and M. K. J. Siddiqui. "Silver nanoparticle applications and human health." Clinica chimica acta 411.23-24 (2010): 1841-1848.

5. Ahmed, Shakeel, et al. "A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: a green expertise." Journal of advanced research 7.1 (2016): 17-28.