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resazurin 快速筛选有机和无机纳米粒子的抗菌活性:潜力、限制和注意事项。

Resazurin rapid screening for antibacterial activities of organic and inorganic nanoparticles: Potential, limitations and precautions.

机构信息

National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.

National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.

出版信息

Anal Biochem. 2022 Jan 15;637:114449. doi: 10.1016/j.ab.2021.114449. Epub 2021 Nov 8.

Abstract

Nanoparticles have been used as antibacterial agents in several products. To optimize their effectiveness, synthesis processes and particle modifications have been developed, creating the need for a rapid screening method to investigate their potencies. Owing to the opacity and insolubility of nanoparticles, a classical method to determine antibacterial activity-such as the minimum inhibitory concentration (MIC), which relies on turbidimetry-might not apply to them. In this study, we demonstrate the potential of a dye (resazurin)-based assay as an indicator of bacterial growth to rapidly screen the antibacterial activities of both organic and inorganic nanomaterials against both gram-negative (E. coli) and gram-positive (S. aureus) bacteria. The results indicate that the resazurin-based assay successfully determine the MIC of organic lipid nanocarriers, and several inorganic nanoparticles. However, the use of resazurin require a precaution for nanoparticles with photocatalytic properties, which may cause dye degradation at higher concentrations. In this study, resazurin bleaching was observed at approximately >50 mg/ml of TiO. In summary, the modified MIC assay with resazurin can evaluate antibacterial activity of nanomaterials, whose turbidity interferer conventional MIC assay. This modification conserves an advantage of MICs assay which are simple and reliable. This would be useful for screening of antibacterial nanomaterials.

摘要

纳米粒子已被应用于多种抗菌产品中。为了提高其效力,人们开发了合成工艺和颗粒改性方法,这就需要一种快速筛选方法来评估它们的效能。由于纳米粒子不透明和不溶性,一种经典的抗菌活性测定方法(如最小抑菌浓度 (MIC),该方法依赖浊度法)可能不适用于它们。在这项研究中,我们展示了一种基于染料(resazurin)的测定法作为细菌生长指示剂的潜力,可用于快速筛选有机和无机纳米材料对革兰氏阴性(大肠杆菌)和革兰氏阳性(金黄色葡萄球菌)细菌的抗菌活性。结果表明,基于 resazurin 的测定法成功地确定了有机脂质纳米载体和几种无机纳米粒子的 MIC。然而,对于具有光催化性能的纳米粒子,使用 resazurin 需要谨慎,因为在较高浓度下可能会导致染料降解。在本研究中,在约 >50mg/ml 的 TiO 时观察到 resazurin 漂白。总之,用 resazurin 改良的 MIC 测定法可以评估纳米材料的抗菌活性,其浊度会干扰传统的 MIC 测定法。这种改良保留了 MIC 测定法的一个优势,即简单可靠。这对于筛选抗菌纳米材料将非常有用。

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