Introspective study of green-synthesized nanomaterials for the delivery of 2c, novel deubiquitinating enzymes inhibitor, for the treatment of cutaneous melanoma

The overall goal of the project was to develop a drug delivery system based on green chemistry for administering a novel deubiquitinating enzyme inhibitor, 2c, to treat advanced melanoma within the iToBoS project framework. Green chemistry in nanotechnology research is a rising demand, promoting economical, eco-friendly approaches by reducing waste and chemical toxins. The project began with synthesizing, characterizing, and functionalizing AuNPs derived from food extracts to bind the drug for administration to 2D melanoma cell cultures, assessing 2c’s effects on cell viability, cytoskeleton, and membranes. Four food extracts (lemon juice, coffee Arabica, tea leaves, cocoa powder) were evaluated, focusing on forming monodisperse spherical AuNPs under 20 nm. Only cocoa powder enabled successful synthesis, yielding 11 nm gold nanoparticles with a protective organic coating. Spectroscopic analyses (UV-Vis, Raman, ATR-FTIR, XPS) clarified the redox reaction and identified catechins as primary reducing and stabilizing agents, with fatty acids and proteins aiding the protective coating’s formation. However, this coating hindered functionalization, leading to a switch to lipid-based nanoparticles, particularly liposomes. Liposomes, optimized for biocompatibility, effectively encapsulated 2c, enhancing solubility, stability, and controlled release, improving therapeutic outcomes by delaying apoptosis. A 3D bioprinting approach was later adopted to mimic the tumor microenvironment, creating melanoma cell-laden hydrogels with bioinks, with and without hyaluronic acid (HA). HA improved hydrogel stability and viscoelasticity but reduced cell metabolic activity, forming an inert matrix limiting melanoma cell proliferation and migration. Without HA, hydrogels lost stiffness due to swelling, whereas HA-containing scaffolds retained rigidity. Cellularized scaffolds showed stiffness reduction due to enzymatic degradation. Drug delivery tests using 2c (free and liposome-loaded) indicated limited effects on proliferation within 48 hours but reduced scaffold stiffness, highlighting its mechanical modulation potential in 3D cultures. This work bridges 2D and 3D systems, demonstrating green nanotechnology’s role in drug delivery development while comparing natural materials' advantages and challenges.