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Effect of capping agents on the cytotoxicity of silver nanoparticles in human normal and cancer skin cell lines

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Abstract

Silver nanoparticles (AgNPs) are among the most widely used nanomaterials in medical and consumer products. However, safety in the uses of AgNPs is still controversial. The toxicity of AgNPs toward various cell types has been reported to depend on the surface properties of the nanoparticles. In this study, the effect of AgNPs with the average size of 5–15 nm on the viability of the CCD-986SK human normal skin fibroblast cell line and A375 human malignant melanoma cell line was evaluated. Comparative toxicity studies, based on MTT assay, were performed by using either sodium alginate or poly (4-styrenesulfonic acid-co-maleic acid) sodium salt (PSSMA) as capping agent in the nanoparticle preparation. The cytotoxicity tests revealed that AgNO3 alone was highly toxic to both cell types while both alginate and PSSMA alone were not toxic. AgNPs capped with alginate were selectively toxic to the cancer cell line but not to the normal cell line while AgNPs capped with PSSMA were toxic to both cancer and normal cell lines. Judging from the 50 % inhibition concentration (IC50), it was found that the cancer cell line was more sensitive to AgNPs than the normal cell line. Study on the mode of cell death by annexin V and propidium iodide staining revealed that AgNPs induced more apoptotic cell death (84–90 %) than necrosis (8–12 %) in the skin cancer cell line. These results suggest that the toxicity of AgNPs depended on the type of capping agent and the type of cell line.

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Acknowledgments

This research was financially supported by Chulalongkorn University, Ratchadaphiseksomphot Endowment Fund (grant no. GF_58_05_61_01), and Royal Thai Government budget (grant no. GBR_BSS_100_59_61_07).

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Authors and Affiliations

  1. Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Ponsawan Netchareonsirisuk

  2. Antibody Production Research Unit, Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, 10330, Thailand

    Songchan Puthong & Kittinan Komolpis

  3. Petroleum and Petrochemical College, Chulalongkorn University, Bangkok, 10330, Thailand

    Stephan Dubas

  4. Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Tanapat Palaga

Authors
  1. Ponsawan Netchareonsirisuk
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  2. Songchan Puthong
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  3. Stephan Dubas
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  4. Tanapat Palaga
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  5. Kittinan Komolpis
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Correspondence to Kittinan Komolpis.

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Electronic supplementary material

Supplementary Fig. 1

UV-Vis absorbance spectra of AgNPs solutions capped with Alginate (0.04–4.56 mM) using NaBH4 at (A) 3.8 mM, (B) 19 mM and (C) 38 mM. Each line represents spectra of the AgNPs solution prepared at different molar ratio of AgNO3 to alginate to NaBH4 (GIF 127 kb)

High-resolution image (TIFF 74933 kb)

Supplementary Fig. 2

UV-Vis absorbance spectra of AgNPs solutions capped with PSSMA (0.04–4.56 mM) using NaBH4 at (A) 3.8 mM, (B) 19 mM and (C) 38 mM. Each line represents spectra of the AgNPs solution prepared at different molar ratio of AgNO3 to PSSMA to NaBH4 (GIF 124 kb)

High-resolution image (TIFF 75731 kb)

Supplementary Fig. 3

Size distribution plots of AgNPs prepared at different ratios of the precursors (GIF 160 kb)

High-resolution image (TIFF 54392 kb)

Supplementary Fig. 4

Cell viability of human skin normal cell (CCD-986SK) (♦) and human skin cancer cell (A375) (▲) after treatment with (a) AgNO3, (b) alginate and (c) PSSMA for 72 h (GIF 168 kb)

High-resolution image (TIFF 64149 kb)

Supplementary Fig. 5

Cytotoxicity of AgNPs capped with (a) alginate and (b) PSSMA at (1) 0.23 mM, (2) 1.14 mM and (3) 4.56 mM to human skin cancer cell (A375) after exposure for 24, 48, and 72 h (GIF 188 kb)

High-resolution image (TIFF 47026 kb)

Supplementary Fig. 6

Annexin V-PI staining of CCD-986SK cells treated with 600 μg/mL AgNPs for 72 h. A: untreated and unstained cells, B: untreated cells stained with Annexin V and PI, C: cells treated with AgNPs capped with alginate and stained with Annexin V and PI, D: cells treated with AgNPs capped with Copss and stained with Annexin V and PI, E: cells treated with 1 μg/mL doxorubicin and stained with Annexin V and PI (positive control). Quadrant 1: necrosis, quadrant 2: late apoptosis, quadrant 3: live cell, quadrant 4: early apoptosis (GIF 227 kb)

High-resolution image (TIFF 8120 kb)

Supplementary Fig. 7

Annexin V-PI staining of A375 cells treated with 600 μg/mL AgNPs for 72 h. A: untreated and unstained cells, B: untreated cells stained with Annexin V and PI, C: cells treated with AbNPs capped with alginate and stained with Annexin V and PI, D: cells treated with AgNPs capped with Copss and stained with Annexin V and PI, E: cells treated with 1 μg/mL doxorubicin and stained with Annexin V and PI (positive control). Quadrant 1: necrosis, quadrant 2: late apoptosis, quadrant 3: live cell, quadrant 4: early apoptosis (GIF 215 kb)

High-resolution image (TIFF 7985 kb)

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Netchareonsirisuk, P., Puthong, S., Dubas, S. et al. Effect of capping agents on the cytotoxicity of silver nanoparticles in human normal and cancer skin cell lines. J Nanopart Res 18, 322 (2016). https://doi.org/10.1007/s11051-016-3624-6

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