Synthesis, functionalization and characterization of Polysaccharide-coated Protein-based Nanoparticles for Curcumin delivery

Over the past decades, nanoparticles (NPs) have garnered significant research interest as drug delivery platforms due to properties such as targeted cell interaction and controlled drug release. In medicine, they show promise, especially in chemotherapy, by enabling precise delivery of therapeutic agents to cancer cells while minimizing damage to healthy tissues. This thesis focuses on the synthesis and characterization of protein-based nanoparticles composed of zein and sodium caseinate (Zein/NaCas), functionalized with polysaccharide coatings to enhance stability. Curcumin is a natural polyphenol derived from Curcuma longa, known for its antioxidant, anti-inflammatory, antimicrobial, and anticancer properties. Despite these benefits, its poor water solubility, instability, and rapid metabolism limit its bioavailability. To overcome these challenges, curcumin was encapsulated in Zein/NaCas nanoparticles, enhancing its solubility, protecting it from degradation, and enabling sustained release. Nanoparticles were synthesized using a pH-driven self-assembly method without organic solvents, ensuring biocompatibility and reproducibility. Various polysaccharide coatings i.e. alginate, chitosan, hyaluronic acid, and a chitosan-hyaluronic acid bilayer, were applied to improve colloidal stability, encapsulation efficiency, and active tumor targeting via interaction with CD44 receptors, often overexpressed in cancer cells. Characterization, including size, polydispersity index (PDI), zeta potential, and stability over time, were measured through Dynamic light scattering (Zetasizer NanoS). Encapsulation efficiency was assessed via UV-Vis spectroscopy (Jasco V650). Vitality of cells was measured via bright field microscope (LasX) by evaluating morphological changes as an indicator of citotoxicity. In vitro tests were performed on the human osteosarcoma U2OS cancer cell line, previously used to assess the efficacy of the antitumoral activity of curcumin. The nanoparticles were also tested on MCF10A cell line to evaluate their specificity towards tumor cells. Overall, this study demonstrates the potential of functionalized proteic nanoparticles as smart drug delivery systems for hydrophobic therapeutic agents, supporting future applications in cancer nanomedicine.