Impact of Graphene Oxide Nanoparticles on In Vitro Development of a Mouse Preimplantation Embryo and Interaction With the Zona Pellucida.

The development of assisted reproductive technologies increases the likelihood of nanoparticles' (NPs) direct contact with gametes and embryos in in vitro conditions. Analyzing the influence of nanomaterials on the early mammalian embryo becomes increasingly relevant. This work is devoted to the effect of graphene oxide (GO) NPs on the in vitro development of mammalian embryos. Mouse 2-cell embryos were preincubated with GO NPs. The interaction of GO with the Zona Pellucida (ZP) of the embryo was investigated using fluorescence lifetime imaging with two-photon excitation (2p-FLIM). During embryo development, the NPs penetration into ZP (blastocyst stage) and perivitelline space (blastocyst hatching stage) was observed. Despite this, GO did not affect the embryo's ability to develop till late and hatching blastocysts. The mechanism of the NPs getting into the perivitelline space and the consequences of NP-embryo direct contact are discussed. The 2p-FLIM efficiency for studying NP interaction with mammalian embryos is evaluated.