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Elucidating the mechanism of cellular uptake

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Currently m NPs are employed in clinic in MRI, which allows intra-tissue and intracellular detection.

The major hurdle presented to cellular internalisation is the cell plasma membrane, which acts as a barrier to block the molecules that are usually not actively imported by cells.

This selective permeability of the cell membrane is due to the presence of the lipid bilayer, which is interspersed with transmembrane proteins.

The hydrophobic nature of these lipids prevents the diffusion of various polar solutes such as proteins and peptides across the cell membrane, leading to the prevention of unconstrained influx and efflux of various solutes [5].

In the study herein, fluorescently tagged 200nm m NPs functionalised with a penetratin were introduced to monolayer fibroblast cells in the presence and absence of a magnetic field.

Cell internalisation was quantified, and the mechanism of uptake was analysed using PCR and western blot (clathrin and caveolin) alongside internalisation when cultured with the specified blockers.

In addition to the use of CPPs, the efficacy of m NP internalisation may be further enhanced by the use of external static magnetic fields [21] through a process termed ‘magnetofection’, which is capable of increasing the transfection rates to about 100 fold using extremely low concentrations of m NPs [22].

We have previously shown that both the use of magnetic field and penetratin increased cellular uptake of 500 nm NPs [23].

It involves the recruitment of the endocytic structures that are stabilized by clathrin triskelia that form the clathrin coat, and contain a N-terminal β-propeller domain (TD) that acts as a hub for protein-protein interaction [8].

Therefore, this paper employs two different blockers of endocytosis, Pitstop 2 and Dyngo 4a, in a bid to determine the cellular uptake route.

Pitstop 2 impedes the receptor-mediated endocytosis by arresting the clathrin-coated pit dynamics at multiple stages [10], whilst Dyngo 4a is a structural analog of Dynasore with increased efficacy and is a potent therapeutic target, for the treatment of Botulism as well as various other diseases involving the dynamin-dependent uptake mechanism [25].

Due to their small sizes, the nanoparticles can cross most of the biological barriers such as the blood vessels and the blood brain barrier, thus providing ubiquitous access to most tissues.

In all biomedical applications maximum nanoparticle uptake into cells is required.