Department of Microbiology, Nicolaus Copernicus University, Lwowska 1, 87100 Torun, Poland.
Department of Immunology, Nicolaus Copernicus University, Lwowska 1, 87100 Torun, Poland.
Molecules. 2020 Jul 2;25(13):3022. doi: 10.3390/molecules25133022.
The development of nanotechnology in the last two decades has led to the use of silver nanoparticles (AgNPs) in various biomedical applications, including antimicrobial, anti-inflammatory, and anticancer therapies. However, the potential of the medical application of AgNPs depends on the safety of their use. In this work, we assessed the in vitro cytotoxicity and genotoxicity of silver nanoparticles and identified biomolecules covering AgNPs synthesized from actinobacterial strain SH11. The cytotoxicity of AgNPs against MCF-7 human breast cancer cell line and murine macrophage cell line RAW 264.7 was studied by MTT assay, cell LDH (lactate dehydrogenase) release, and the measurement of ROS (reactive oxygen species) level while genotoxicity in cells was testing using the Ames test. The in vitro analysis showed that the tested nanoparticles demonstrated dose-dependent cytotoxicity against RAW 264.6 macrophages and MCF-7 breast cancer cells. Moreover, biosynthesized AgNPs did not show a mutagenic effect of . The analyses and identification of biomolecules present on the surface of silver nanoparticles showed that they were associated with proteins. The SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) analysis revealed the presence of 34 and 43 kDa protein bands. The identification of proteins performed by using LC-MS/MS (liquid chromatography with tandem mass spectrometry) demonstrated their highest homology to bacterial porins. Capping biomolecules of natural origin may be involved in the synthesis process of AgNPs or may be responsible for their stabilization. Moreover, the presence of natural proteins on the surface of bionanoparticles eliminates the postproduction steps of capping which is necessary for chemical synthesis to obtain the stable nanostructures required for application in medicine.
在过去的二十年中,纳米技术的发展导致了银纳米粒子(AgNPs)在各种生物医学应用中的使用,包括抗菌、抗炎和抗癌疗法。然而,AgNPs 医学应用的潜力取决于其使用的安全性。在这项工作中,我们评估了银纳米粒子的体外细胞毒性和遗传毒性,并鉴定了覆盖从放线菌菌株 SH11 合成的 AgNPs 的生物分子。通过 MTT 测定、细胞 LDH(乳酸脱氢酶)释放和 ROS(活性氧)水平的测量研究了 AgNPs 对 MCF-7 人乳腺癌细胞系和 RAW 264.7 鼠巨噬细胞系的细胞毒性,而细胞遗传毒性则使用 Ames 试验进行测试。体外分析表明,测试的纳米粒子对 RAW 264.6 巨噬细胞和 MCF-7 乳腺癌细胞表现出剂量依赖性的细胞毒性。此外,生物合成的 AgNPs 没有显示出诱变作用。对银纳米粒子表面存在的生物分子的分析和鉴定表明,它们与蛋白质有关。SDS-PAGE(十二烷基硫酸钠-聚丙烯酰胺凝胶电泳)分析显示存在 34 和 43 kDa 的蛋白质带。使用 LC-MS/MS(液相色谱-串联质谱)进行的蛋白质鉴定表明它们与细菌孔蛋白具有最高的同源性。天然来源的封端生物分子可能参与 AgNPs 的合成过程,或者可能负责其稳定性。此外,生物纳米粒子表面存在天然蛋白质消除了化学合成获得用于医学应用所需的稳定纳米结构所需的后生产封端步骤。