Molecular dynamics simulations of cRGD-conjugated PEGylated TiO$_2$ nanoparticles for targeted photodynamic therapy

Abstract: The conjugation of high-affinity cRGD-containing peptides is a promising approach in nanomedicine to efficiently reduce off-targeting effects and enhance the cellular uptake by integrin-overexpressing tumor cells. Herein we utilize atomistic molecular dynamics simulations to evaluate key structural-functional parameters of these targeting ligands for an effective binding activity towards $\alpha_V\beta_3$ integrins. An increasing number of cRGD ligands is conjugated to PEG chains grafted to highly curved TiO$_2$ nanoparticles to unveil the impact of cRGD density on the ligand's presentation, stability, and conformation in an explicit aqueous environment. We find that a low density leads to an optimal spatial presentation of cRGD ligands out of the "stealth" PEGylated layer around the nanosystem, favoring a straight upward orientation and spaced distribution of the targeting ligands in the bulk-water phase. On the contrary, high densities favor clustering and internalization of cRGD ligands in the inner region of the PEGylated layer, driven by a concerted mechanism of enhanced ligand-ligand interactions and reduced water accessibility over the ligand's molecular surface. These findings strongly suggest that the ligand density modulation is a key factor in the design of cRGD-targeting nanodevices to maximize their binding efficiency into over-expressed $\alpha_V\beta_3$ integrin receptors.