Abstract
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 Salmonella typhimurium 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 S. typhimurium. 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.
Highlights
Nanotechnology and related nanomaterials have become the most leading and promising areas for scientific research and technological development
UV-vis, TEM and zeta potential analyses confirmed the synthesis of silver nanoparticles from actinobacterial strain
Analyses showed that the biosynthesized AgNPs demonstrated dose-dependent cytotoxicity in both tested cell lines, the macrophages were more sensitive to AgNP treatment than MCF-7 cancer cells
Summary
Nanotechnology and related nanomaterials have become the most leading and promising areas for scientific research and technological development. The creation and utilization of nanomaterials and nanodevices already have multiple applications in various fields [1]. Nanomedicine brings nanotechnology and medicine together, in order to develop novel diagnostics, therapies and improve existing treatments [2]. Recent advances in nanotechnology have broadened the potential applications for silver nanoparticles (AgNPs), at the same time increasing human exposure to these materials and raising anxiety regarding their adverse biological effects [3]. The synthesis of nanoparticles (NPs) may be performed by chemical, physical, and biological approaches. The biological synthesis results in high yield, good solubility, high stability and biocompatibility of obtained nanoparticles [6]. In the extracellular synthesis of silver nanoparticles, the reducing agents, including amino acids, membrane proteins, NAD(P)+ reductases, dehydrogenases and various secondary metabolites, are involved, whereas the nanoparticle capping agents are formed by extracellular proteins, enzymes or peptides [7,8]
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