Nano-Ag Particles Embedded in C-Matrix: Preparation, Properties and Application in Cell Metabolism

Abstract

The application of nano-Ag grains as antiviral and antibacterial materials is widely known since ancient times. The problem is the toxicity of the bulk or big-size grain materials. It is known that nano-sized silver grains affect human and animal cells in some medical treatments. The aim of this study is to investigate the influence of nano-Ag grains embedded in a carbonaceous matrix on cytotoxicity, genotoxicity in fibroblasts, and mutagenicity. The nanocomposite film is composed of silver nanograins embedded in a carbonaceous matrix and it was obtained via the PVD method by deposition from two separated sources of fullerenes and silver acetate powders. This method allows for the preparation of material in the form of a film or powder, in which Ag nanograins are stabilized by a carbon network. The structure and morphology of this material were studied using SEM/EDX, XRD, and Raman spectroscopy. The toxicology studies were performed for various types of the material differing in the size of Ag nanograins. Furthermore, it was found that these properties, such as cell viability, genotoxicity, and mutagenicity, depend on Ag grain size.

Keywords: cytotoxicity; genotoxicity; mutagenicity; nanocomposite nano-Ag.

Figure 1

(a) SEM image of surface of C-nAg film on alumina substrate (equivalent to Si substrate), (b) EDX spectrum from this sample, (c) SEM image of cross-section of the film deposited on Si substrate, (d) statistical analysis of Ag nanograins size distribution.

Figure 2

X-ray diffraction patterns for: (a) The P1 sample (red line); (b) the P2 sample (blue line).

Figure 3

Raman spectrum of the prepared nano-Ag-C film deposited on alumina substrate.

Figure 4

The result of the decomposition of the Raman spectrum of nano-Ag-C film in the area of 800–2200 cm⁻¹. The line colors are as follows: Black line—experimental spectrum; brown line—calculated spectrum; other colors—arbitrary assigned by Peak Spectroscopy Software.

Figure 5

Cytotoxic effect of P1 and P2 in L929 cell line, detected with the MTT reduction assay (A,B), with the NRU assay (C,D), and with the LDH release assay (E,F). * p < 0.05, significance of difference compared with control.

Figure 5

Cytotoxic effect of P1 and P2 in L929 cell line, detected with the MTT reduction assay (A,B), with the NRU assay (C,D), and with the LDH release assay (E,F). * p < 0.05, significance of difference compared with control.

Figure 6

Typical fluorescence microscopic images of L929 cells exposed to P1 (A) and P2 (B) and comets formed after exposure of P1 (C).

Figure 7

Fluorescence microscopic images (micronuclei) of the acridine orange stained L929 cells exposed to P1 (A) and P2 (B).