The investigations will be devoted to cellular uptake of nanoscale structures and the influence of the protein corona on the phenomenon. Initially, a library of coumarin-6 loaded polymeric nanoparticles will be manufactured from the pH-responsive and biocompatible block copolymers PEOm-b-PDPAn, PMPCm-b-PDPAn e PHPMAm-b-PDPAn. They will be further deeply characterized by using essentially scattering techniques (dynamic light scattering - DLS, static light scattering - SLS and electrophoretic light scattering - ELS) besides atomic force microscopy - AFM. Subsequently it will be evaluated the cellular uptake of the produced entities and the relation with their structural features, particularly the nature of the hydrophilic stabilizing shell (PEO, PMPC and PHPMA). These investigations will be performed using the resources of flow cytometry (FACS) and laser scanning confocal microscopy (LSCM). Afterwards, the focus will be moved to the protein corona formation. The produced nanoparticles will be incubated in plasma environment and by using centrifugation, Polyacrylamide gel electrophoresis (SDS-PAGE) and Liquid chromatography-mass spectrometry (LC-MS) it will be identified and quantified the adsorbed macromolecules in distinct entities. The cellular uptake of the protein-nanoparticle complexes will be further investigated in order to shed some light on its effect on the cell internalization process. Finally, similar evaluation will be conducted in the surface functionalized entity RGD-PHPMAm-b-PDPAn. The entity will be functionalized by using the peptide sequence Arginylglycylaspartic acid (RGD). This is a tripeptide composed of L-arginine, glycine, and L-aspartic acid and the sequence is a common element in cellular recognition.
News published in Agência FAPESP Newsletter about the scholarship: