In vitro study of silver nanoparticles-induced embryotoxicity using a rat whole embryo culture model

Recently, our in vivo experiment showed that silver nanoparticles (AgNPs) did not cause developmental toxicity. However, the putative influences of direct exposure of AgNPs on the embryo-fetuses could not be elucidated because the embryo-fetus was exposed to AgNPs through their dams. In this study, the potential impact of AgNPs on embryonic development during the critical phase of organogenesis was examined utilizing a rat whole embryo culture model. This system could separate the direct effects of AgNPs from those that are maternally mediated. To evaluate the embryotoxic potential of AgNPs, embryos were exposed to 1.67, 5, and 15 μg/mL of AgNPs for 48 h. At the conclusion of the culture period, embryonic growth and development were assessed, and morphological abnormalities were systematically evaluated. Also, apoptosis induced by AgNPs was evaluated by TUNEL and immunohistochemistry for caspase-3. At 15 μg/mL, a retardation in embryonic growth and differentiation, accompanied by a heightened frequency of morphological abnormalities, including abnormal axial rotation, open neural tube, absent optic vesicle, and growth retarded were observed in a dose-dependent manner. At this concentration, caspase-3-positive cells appeared in the treated embryonic tissues compared to controls. At 5 μg/mL, AgNPs also caused a decrease in the embryonic otic system, somite number, and total morphological score. No adverse effects on embryonic growth and development associated with the treatment were observed at 1.67 μg/mL. The findings demonstrated that the direct exposure of AgNPs to rat embryos induces developmental delays and morphological abnormalities, and that AgNPs can induce a direct developmental toxicity and caspase-dependent apoptosis in rat embryos.