Silver nanoparticle induced toxicity and cell death mechanisms in embryonic zebrafish cells

Abstract

From their first contact with continuously cultured ZF4 cells, 10, 30 and 100 nm PVP-capped AgNPs induce receptor mediated endocytotic processes, cellular repair mechanisms, and at higher doses, cell death via autophagy, apoptosis or necrosis.

Cellular repair mechanisms triggered by exposure to silver nanoparticles and ionic silver in embryonic zebrafish cells

“…ZF4 cells established from 1 day old zebrafish embryo were purchased from ATTC (https://www.atcc.org/products/all/CRL-2050.aspx) and cultured as described on the manufacturer's website and in ref. 22. Briefly, ZF4 cells were cultured in vented cap T75 flasks (Corning, CLS430641) in a total volume of 10 mL of CCM and incubated in a humidified atmosphere of 5% CO 2 at 28°C.…”

“…24 This modified protocol was selected due to recorded interferences from the absorbance properties of the AgNPs when applying the manufacturer's protocol. [24][25][26] The methodology is described on cancer cells in the original publication by Ali-Boucetta et al (2011), 24 and in a previous ZF4 cell publication by Quevedo et al (2021). 22 Briefly, ZF4 cells were seeded in 96well plates at a density of 8000 cells in a total volume of 200 μL per well in CCM or SFM and incubated at 28°C and 5% CO 2 .…”

“…[24][25][26] The methodology is described on cancer cells in the original publication by Ali-Boucetta et al (2011), 24 and in a previous ZF4 cell publication by Quevedo et al (2021). 22 Briefly, ZF4 cells were seeded in 96well plates at a density of 8000 cells in a total volume of 200 μL per well in CCM or SFM and incubated at 28°C and 5% CO 2 . The next day, cells were treated with AgNPs or AgNO 3 in either CCM (10% FBS) for 3, 24, 48 and 72 hours or SFM for 3, 6, 12, and 24 hours.…”

“…Further characterisation of the NPs using different concentrations and testing media is broadly described in a previous publication. 22…”

“…† For more information about other tested concentrations, exposure time points and the full methodology used for treatments prepared in CCM, see Quevedo et al, (2021). 22 To understand the cytotoxic effects of the AgNPs and its ionic counterpart under serum free conditions, the EC 50 in ZF4 cells was also determined. The AgNP concentrations in SFM were equal to those in the CCM treatments; however, due to the high toxicity recorded (reduction in cell viability %) after 24 hours (<10% viability) results are displayed only up to 24 hours of incubation (Fig.…”