Biosynthesis of gold nanoparticles by two bacterial and fungal strains, Bacillus cereus   and Fusarium oxysporum  , and assessment and comparison of their nanotoxicity in vitro by direct and indirect assays

Background: Although nanoparticles (NPs) have many advantages, it has been proved that they may be absorbed by and have toxic effects on the human body. Recent research has tried to evaluate and compare the nanotoxicity of gold nanoparticles (AuNPs) produced by two types of microorganisms in vitro by two different methods. AuNPs were produced by Bacillus cereus and Fusarium oxysporum, and their production was confirmed by visible spectral, transmission electron microscope, and X-ray diffraction (XRD) analyses. The human fibroblast cell line CIRC-HLF was treated with AuNPs, and the induced nanotoxicity was measured using direct microscopic and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays.
Results: The results showed that the produced AuNPs had a maximum absorbance peak around 510–530 nanometer (nm), with spherical, hexagonal, and octagonal shapes and average sizes around 20–50 nm. The XRD results confirmed the presence of GNPs in the microbial culture supernatants. An MTT assay showed that GNPs had dose-dependent toxic effects, and microscopic analysis showed that GNPs induced cell abnormalities in doses lower than the determined half-maximal inhibitory concentrations (IC50s).
Conclusions: In conclusion, the biologically produced AuNPs had toxic effects in the cell culture, and direct techniques such as microscopic evaluation instead of indirect methods such as MTT assay were more useful for assessing the nanotoxicity of the biologically produced AuNPs. Thus, the use of only MTT assay for nanotoxicity evaluation of AuNPs is not desirable.

Keywords
Biologically produced nanoparticles; Drug delivery; In vitro nanotoxicity; In vivo nanotoxicity; Nanoparticle production; Nanotechnology; Nanotoxicity analysis; Purification of nanoparticles; Surface plasmon resonance; Visible spectral analysis; X-ray diffraction analysis