Abstract
Glioblastomas (GBM) are aggressive brain tumors with very poor prognosis. While silver nanoparticles represent a potential new strategy for anticancer therapy, the silver/silver chloride nanoparticles (Ag/AgCl-NPs) have microbicidal activity, but had not been tested against tumor cells. Here, we analyzed the effect of biogenically produced Ag/AgCl-NPs (from yeast cultures) on the proliferation of GBM02 glioblastoma cells (and of human astrocytes) by automated, image-based high-content analysis (HCA). We compared the effect of 0.1–5.0 µg mL−1 Ag/AgCl-NPs with that of 9.7–48.5 µg mL−1 temozolomide (TMZ, chemotherapy drug currently used to treat glioblastomas), alone or in combination. At higher concentrations, Ag/AgCl-NPs inhibited GBM02 proliferation more effectively than TMZ (up to 82 and 62% inhibition, respectively), while the opposite occurred at lower concentrations (up to 23 and 53% inhibition, for Ag/AgCl-NPs and TMZ, respectively). The combined treatment (Ag/AgCl-NPs + TMZ) inhibited GBM02 proliferation by 54–83%. Ag/AgCl-NPs had a reduced effect on astrocyte proliferation compared with TMZ, and Ag/AgCl-NPs + TMZ inhibited astrocyte proliferation by 5–42%. The growth rate and population doubling time analyses confirmed that treatment with Ag/AgCl-NPs was more effective against GBM02 cells than TMZ (~ 67-fold), and less aggressive to astrocytes, while Ag/AgCl-NP + TMZ treatment was no more effective against GBM02 cells than Ag/AgCl-NPs monotherapy. Taken together, our data indicate that 2.5 µg mL−1 Ag/AgCl-NPs represents the safest dose tested here, which affects GBM02 proliferation, with limited effect on astrocytes. Our findings show that HCA is a useful approach to evaluate the antiproliferative effect of nanoparticles against tumor cells.
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Acknowledgements
This work was supported by the National Council for Scientific and Technological Development (CNPq), the Carlos Chagas Filho Foundation for Research Support of the State of Rio de Janeiro (FAPERJ) and the Coordination for the Improvement of Higher Education Personnel (CAPES).
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Supplementary Fig.
1. Representative images showing the effect of Ag/AgCl-NPs on the proliferation of GBM02 cells and astrocytes. Cells labeled with Hoechst 33258 were left untreated, or were treated with different concentrations of Ag/AgCl-NPs (0.1, 0.5, 1.0, 2.5 and 5.0 µg mL-1) for 72 h. Images are representative of 3 independent experiments (6 fields imaged per experimental condition, in each experiments). Scale bar: 50 µm. (TIFF 20027 kb)
Supplementary Fig.
2. Representative images showing the effect of TMZ on the proliferation of GBM02 cells and astrocytes. Cells labeled with Hoechst 33258 were left untreated, or were treated with different concentrations of TMZ (9.7, 19.4, 29.1, 38.5 and 48.5 µg mL−1) for 72 h. Images are representative of 3 independent experiments (6 fields imaged per experimental condition, in each experiments). Scale bar: 50 µm. (TIFF 20027 kb)
Supplementary Fig.
3. Representative images showing the effect of Ag/AgCl-NPs + TMZ on the proliferation of GBM02 cells and astrocytes. Cells labeled with Hoechst 33258 were left untreated, or were treated with different concentrations of Ag/AgCl-NPs + TMZ (0.1 + 9.7, 0.5 + 19.4, 1.0 + 29.1, 2.5 + 38.5 and 5.0 + 48.5 µg mL−1) for 72 h. Images are representative of 3 independent experiments (6 fields imaged per experimental condition, in each experiments). Scale bar: 50 µm. (TIFF 18706 kb)
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Eugenio, M., Campanati, L., Müller, N. et al. Silver/silver chloride nanoparticles inhibit the proliferation of human glioblastoma cells. Cytotechnology 70, 1607–1618 (2018). https://doi.org/10.1007/s10616-018-0253-1
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DOI: https://doi.org/10.1007/s10616-018-0253-1