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SCCS/1596/18 Final Opinion on Colloidal Silver (nano) __________________________________________________________________________________________ *Evaluation of genotoxicity was impossible due to the observed cytotoxic effect of the tested sample. The conclusion from the study is that within the tested scope, the sample is non-genotoxic. Without metabolic activation, cytotoxicity of the tested sample has been observed. Ref.: 59 SCCS comment: The information provided in the additional study on cytotoxicity and genotoxicity is not acceptable, nor sufficient. The results of the whole study (on both cytotoxicity and genotoxicity testing) are not reliable for the following reasons: SCCS comments on the cytotoxic study: - According to the data provided, only one concentration of 50 ppm (50 μg/mL) was tested and no cytotoxicity was observed. However, from the information available in published literature, it is known that the EC50 for silver nanoparticles may vary and can be below 50 μg/mL, depending on cell types and particle sizes (Liu et al, 2010, Braydich-Stolle et al, 2005, Rosario et al, 2016, Park et al, 2011, Huk et al, 2014). - No information on control substances used was given, neither positive nor negative. - No data are provided on stability of the silver nanoparticle suspension and how it was applied on the agar. - No information on number of replicates is given. - The agar diffusion test used is not considered suitable to determine cytotoxic properties of silver nanoparticles. According to PN-EN ISO 10993-5:2009 ('8.4.1 Agar diffusion 8.4.1.1) the test allows only a qualitative assessment of cytotoxicity. Also, ISO 10993-5 is dedicated mainly to the testing of extracts of medical devices and not pure chemicals. - More specifically for nanomaterials, ISO 19007 describes an in-vitro MTS assay for measuring cytotoxic effects of nanomaterials. Other quantitative assessments also might be used (such as the NRU cytotoxicity test; the colony formation cytotoxicity test; the MTT cytotoxicity test and the XTT cytotoxicity test) under the condition that assay interference is considered. The SCCS is therefore of the opinion that a method that is not prone to interference should be preferably used, such as colony-forming efficacy. - The cytotoxicity test should be carried out at different concentrations to enable calculation of EC50 to compare the relative toxicity of the various colloidal silver dispersions in nano form. SCCS comments on the genotoxicity study: Ref.: 61, 62, 63, 64, 65 25PDF Image | Consumer Safety SCCS OPINION ON Colloidal Silver (nano)
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