Abstract
The interaction of single-layer graphene oxide (SLGO) and multi-layered
graphene oxide (MLGO) with a cell culture medium (Le. DMEM) was studied
by evaluating fetal bovine serum (FBS) protein corona formation towards
in vitro nanotoxicity assessment and nanobiointeractions. SLGO and MLGO
exhibited different colloidal behavior in the culture medium, which was
visualized by cryogenic transmission electron microscopy in situ
analysis. Exploring proteomics and bioinformatics tools, 394 and 290
proteins were identified on the SLGO and MLGO hard corona compositions,
respectively. From this amount, 115 proteins were exclusively detected
on the SLGO and merely 11 on MLGO. SLGO enriched FBS proteins involved
in metabolic processes and signal transduction, while MLGO enriched
proteins involved in cellular development/structure, and lipid
transport/metabolic processes. Such a distinct corona profile is due to
differences on surface chemistry, aggregation behavior and the surface
area of GO materials. Hydrophilic interactions were found to play a
greater role in protein adsorption by MLGO than SLGO. Our results point
out implications for in vitro studies of graphene oxide materials
concerning the effective dose delivered to cells and corona bioactivity.
Finally, we demonstrated the importance of integrating conventional and
modern techniques thoroughly to understand the GO-FBS complexes towards
more precise, reliable and advanced in vitro nanotoxicity assessment.
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