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Protein adsorption onto Fe3O4 nanoparticles with opposite surface charge and its impact on cell uptake

Nanoparticles (NPs) engineered for biomedical applications are meant to be in contact with protein-rich physiological fluids. These proteins are usually adsorbed onto the NP surface, forming a swaddling layer called protein corona that influences cell internalization. We present a study on protein adsorption onto different magnetic NPs (MNPs) when immersed in cell culture medium, and how these changes affect the cellular uptake. Two colloids with magnetite cores of 25 nm, same hydrodynamic size and opposite surface charge were in situ coated with (a) positive polyethyleneimine (PEI-MNPs) and (b) negative poly(acrylic acid) (PAA-MNPs). After few minutes of incubation in cell culture medium the wrapping of the MNPs by protein adsorption resulted in a 5-fold size increase. After 24 h of incubation large MNP-protein aggregates with hydrodynamic sizes 1500 to 3000 nm (PAA-MNPs and PEI-MNPs respectively) were observed. Each cluster contained an estimated number of magnetic cores between 450 and 1000, indicating the formation of large aggregates with a "plum pudding" structure of MNPs embedded into a protein network of negative surface charge irrespective of the MNP_core charge. We demonstrated that PEI-MNPs are incorporated in much larger amounts than the PAA-MNPs units. Quantitative analysis showed that SH-SY5Y cells can incorporate 100 per cent of the added PEI-MNPs up to about 100 pg per cell, whereas for PAA-MNPs the uptake was less than 50 percent. The final cellular distribution showed also notable differences regarding partial attachment to the cell membrane. These results highlight the need to characterize the final properties of MNPs after protein adsorption in biological media, and demonstrate the impact of these properties on the internalization mechanisms in neural cells.