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细菌介导的具有抗菌、抗氧化和抗癌潜力的氧化铜纳米颗粒的合成与表征。

Bacterial-mediated synthesis and characterization of copper oxide nanoparticles with antibacterial, antioxidant, and anticancer potentials.

作者信息

Talebian Seyedehsaba, Shahnavaz Bahar, Nejabat Masoud, Abolhassani Yasaman, Rassouli Fatemeh B

机构信息

Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.

Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran.

出版信息

Front Bioeng Biotechnol. 2023 Mar 6;11:1140010. doi: 10.3389/fbioe.2023.1140010. eCollection 2023.

Abstract

The application of novel bacterial strains for effective biosynthesis of nanoparticles minimizes negative environmental impact and eliminates challenges of available approaches. In the present study, cell-free extract of sp. BS95. was used for synthesis of copper oxide nanoparticles (CuONPs). Characterization of crude and calcined CuONPs was carried out by UV-vis spectroscopy, X-ray diffraction (XRD), fourier transform infrared (FTIR) spectroscopy, zeta potential, dynamic light scattering, field emission scanning electron microscopy, transmission electron microscopy, and atomic force microscopy. Afterward, biogenic CuONPs were evaluated for antibacterial, antioxidant, and cytotoxic effects using broth micro-dilution method, DPPH assay and alamarBlue assay, respectively. Finally, molecular mechanisms behind anticancer effects of CuONPs was ascertained by real time PCR. UV-vis absorbance spectra registered surface plasmon resonance peaks at 286 nm and 420 nm for crude and calcined CuONPs, respectively. FTIR spectra exhibited bands associated with organic functional groups of bacterial proteins, confirming capping and functionalization of CuONPs. The average crystallite size of crude and calcined CuONPs was determined as 18.24 and 21.3 nm by XRD, respectively. The average zeta potentials of crude and calcined CuONPs were as -28.57 ± 5.13 and -29.47 ± 4.78 mV, respectively, indicating their high stability. Electron microscopy revealed that crude and calcined CuONPs were roughly spherical particles with an average size of 35.24 ± 4.64 and 43.68 ± 2.31 nm, respectively. Biogenic CuONPs induced antibacterial effects with minimal inhibitory concentrations ranging from 62.5 to 1,000 μg/ml against Gram-negative and Gram-positive strains. The antioxidant activity of crude and calcined CuONPs was found to be 83% ± 2.64% and 78% ± 1.73%, respectively. More intriguingly, CuONPs exerted considerable cytotoxic effects on human colon and gastric adenocarcinoma cells, while induced low toxicity on normal cells. Anticancer effects of biogenic CuONPs were confirmed by significant changes induced in the expression of apoptosis-related genes, including , , and . Hence, biosynthesized CuONPs could be considered as potential antimicrobial, antioxidant and anticancer agents.

摘要

应用新型细菌菌株进行纳米颗粒的有效生物合成可将负面环境影响降至最低,并消除现有方法所面临的挑战。在本研究中,使用了sp. BS95的无细胞提取物来合成氧化铜纳米颗粒(CuONPs)。通过紫外可见光谱、X射线衍射(XRD)、傅里叶变换红外(FTIR)光谱、zeta电位、动态光散射、场发射扫描电子显微镜、透射电子显微镜和原子力显微镜对粗制和煅烧后的CuONPs进行了表征。随后,分别使用肉汤微量稀释法、DPPH测定法和alamarBlue测定法评估了生物合成的CuONPs的抗菌、抗氧化和细胞毒性作用。最后,通过实时PCR确定了CuONPs抗癌作用背后的分子机制。紫外可见吸收光谱显示,粗制和煅烧后的CuONPs在286 nm和420 nm处分别出现表面等离子体共振峰。FTIR光谱显示出与细菌蛋白质有机官能团相关的谱带,证实了CuONPs的封端和功能化。通过XRD测定,粗制和煅烧后的CuONPs的平均微晶尺寸分别为18.24和21.3 nm。粗制和煅烧后的CuONPs的平均zeta电位分别为-28.57±5.13和-29.47±4.78 mV,表明它们具有很高的稳定性。电子显微镜显示,粗制和煅烧后的CuONPs均为大致球形颗粒,平均尺寸分别为35.24±4.64和43.68±2.31 nm。生物合成的CuONPs对革兰氏阴性和革兰氏阳性菌株均具有抗菌作用,最低抑菌浓度范围为62.5至1000μg/ml。发现粗制和煅烧后的CuONPs的抗氧化活性分别为83%±2.64%和78%±1.73%。更有趣的是,CuONPs对人结肠和胃腺癌细胞具有显著的细胞毒性作用,而对正常细胞的毒性较低。生物合成的CuONPs的抗癌作用通过凋亡相关基因(包括 、 、 和 )表达的显著变化得到证实。因此,生物合成的CuONPs可被视为潜在的抗菌、抗氧化和抗癌剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba57/10025390/93ed87a3261a/fbioe-11-1140010-g001.jpg

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