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色素包覆银纳米颗粒对耐药菌株的生物制造及其对人表皮样喉癌(HEp-2)细胞的细胞毒性作用

Bio-fabrication of pigment-capped silver nanoparticles encountering antibiotic-resistant strains and their cytotoxic effect towards human epidermoid larynx carcinoma (HEp-2) cells.

作者信息

Muthukrishnan Lakshmipathy, Chellappa Muralidharan, Nanda Anima, Thukkaram Sudhakar, Selvaraj Gracyfathima, Muthiah Bavanilatha, Sagadevan Suresh, Lett J Anita

机构信息

Leather Process Technology, Tannery Division, CSIR - Central Leather Research Institute (CLRI) Adyar Chennai Tamilnadu 600 020 India

Faculty of Bio & Chemical Engineering, Sathyabama Institute of Science and Technology (Deemed to be University) Rajiv Gandhi Salai Chennai Tamilnadu 600119 India.

出版信息

RSC Adv. 2019 May 21;9(28):15874-15886. doi: 10.1039/c9ra01072f. eCollection 2019 May 20.

Abstract

Bacterial biomolecule-mediated nanoparticle (NP) synthesis constitutes a reliable, eco-friendly approach that ameliorates green-chemistry principles. In this study, stable silver nanoparticles were synthesized by exposing aqueous silver ions to an extracellular diffusible pigment produced by (PA6) under optimized laboratory conditions. Spectroscopic and microscopic analyses showed the typical characteristics of silver with an average size of ∼28.30 nm and spherical shape. The particles were polydispersed and showed no definite agglomeration with a zeta potential of -32.3 mV, conferring stability. Antimicrobial studies were carried out using 5, 15, 25 and 50 μg mL concentrations of pcAgNPs, which showed significant antibacterial activity toward clinically important pathogens at all concentrations compared to with the control sample. The bactericidal effect induced by pcAgNPs associated with cell damage was well demonstrated using electron microscopic studies. ROS production was measured using the DCFH-DA method and the oxidative stress was assessed by measuring the reduced glutathione (GSH) levels. Cytotoxicity studies on HEp-2 (Human Epidermoid Larynx Carcinoma) cells exposed to pcAgNPs showed dose-dependent cytotoxic effect with IC of 14.8 μg mL compared to with IC of 7.38 μg mL for the Vero cell control. Mechanistically, the pcAgNPs activated p53 that induced catalase, leading to apoptosis and DNA fragmentation a p53 transcriptional pathway and electron transport arrest, which resulted in cell death. This synergistic efficacy of pigment-AgNPs demonstrated excellent antimicrobial and anti-proliferative activities, providing a potential lead for developing a broad-spectrum antibacterial agent and improving the therapeutic modalities targeting carcinoma cells at the gene level.

摘要

细菌生物分子介导的纳米颗粒(NP)合成是一种可靠、环保的方法,符合绿色化学原则。在本研究中,在优化的实验室条件下,通过将水相银离子暴露于由(PA6)产生的细胞外可扩散色素中来合成稳定的银纳米颗粒。光谱和显微镜分析显示了银的典型特征,平均尺寸约为28.30 nm,呈球形。颗粒多分散,无明显团聚,zeta电位为-32.3 mV,具有稳定性。使用浓度为5、15、25和50 μg/mL的pcAgNPs进行抗菌研究,结果表明,与对照样品相比,所有浓度的pcAgNPs对临床重要病原体均具有显著的抗菌活性。通过电子显微镜研究充分证明了pcAgNPs诱导的与细胞损伤相关的杀菌作用。使用DCFH-DA方法测量ROS产生,并通过测量还原型谷胱甘肽(GSH)水平评估氧化应激。对暴露于pcAgNPs的HEp-2(人表皮样喉癌)细胞进行的细胞毒性研究显示出剂量依赖性细胞毒性作用,IC为14.8 μg/mL,而Vero细胞对照的IC为7.38 μg/mL。从机制上讲,pcAgNPs激活了诱导过氧化氢酶的p53,导致凋亡和DNA片段化——这是一条p53转录途径和电子传递阻滞,最终导致细胞死亡。色素-AgNPs的这种协同功效表现出优异的抗菌和抗增殖活性,为开发广谱抗菌剂和在基因水平上改善针对癌细胞的治疗方式提供了潜在的线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6a69/9064322/ba9269791b8e/c9ra01072f-f1.jpg

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