Innovative drug-loaded nanoconstruct for theranostic applications against pancreatic cancer

In the past few years, progresses in nanomedicine towards cancer treatment have exhibited that the use of nanoparticles (NPs) can be truly promising, since they have shown to be capable of damaging and killing cancer cells throughout different mechanisms. While various nanomaterials have been de-veloped for this purpose, their use brings with it several potential drawbacks which are still scarcely considered, such as the non-specific uptake of these materials in healthy organs and their potential immunogenicity. This Master Thesis work aims to develop an innovative theranostic immuno-compatible nanoconstruct based on Zinc Oxide nanocrystals (ZnO-NCs), whose properties have revealed huge potential for in vitro and in vivo applications, focusing on one of the most lethal diseases, pancreatic cancer. The nanoparti-cles’ therapeutic potential is due to their biocompatibility and pH-sensitivity to ensure targeted drug release, while, for diagnostic purposes, ZnO-NCs doped with rare earth elements (e.g., Gadolinium and Manganese) can be tested as contrast agent in magnetic resonance imaging (MRI). Therefore, doped and undoped ZnO-NCs were synthesized via the co-precipitation method, characterized in terms of Zeta potential, hydrodynamic diameter, morphology, chemical structure and crystallinity, and functionalized to expose NH2 groups. The study is mostly focused on Gd-doped nanoparticles, loaded with Gemcitabine, which is the reference drug for pancreatic cancer treatment. The drug loading was obtained by adsorption on their surface, and the nanocrystals were finally coated exploiting the freeze thaw technique with a lipidic shell consisting in lipids originating from extracellular vesicles (EVs), together with commercially available lipids coupled with functional peptides (e.g., pancreatic pathological microenvironment homing CKAAKN peptide) for targeting purposes. The obtained nanoconstruct was further characterized by Dynamic Light Scattering, Nanoparticle Tracking Analysis and Fluorescence Microscopy, and tested on two different pancreatic cancer cell lines (i.e. BxPC-3 and AsPC-1) in terms of cellular uptake, cytotoxicity, and apoptotic events. Collaterally, a slightly different nanoconstruct was developed, consisting in the same core of ZnO Gd-doped nanocrystals, but loaded with a poorly hydrophilic drug, i.e. Sorafenib, and coated employing only the above mentioned commer-cially available lipids. This nanoconstruct was then characterized and tested on pancreatic cancer cells similarly to the Gemcitabine-loaded one. The preliminary in vitro results indicate that both these innovative nanocon-structs can be extremely impactful on the standard treatment of pancreatic cancer.