• Biomaterials
• crispr-cas9
• genetic engineering
• nanoparticle
• pancreatic cancer
• scaffold
• tissue engineering

The cellular microenvironment is a network of structural and functional components that provide mechanical and chemical stimuli, which influence cell function and fate.

For this purpose cancer tissue engineering has start to emerge with the aim of understanding the biophysical features that affect cell growth to design biomimetic scaffolds which could be able to stimulate the growing of tumors cells into them to reproduce tumor-tissue like construct as alternative to 2D cells cultures and organoids.
Differently from these, 3D tissue models are able to reproduce the extracellular matrix

Aim of the present thesis is the design, developement and characterization of 3D biobased scaffolds to recreate a more realistic model of the pancreatic ductal adenocarcinoma.
These models are achieved by culturing the cells on these three-dimensional porous structures made of biocompatible materials.
For this purpose, spongy scaffolds made of:

-Polyvinyl alcohol (PVA) and gelatin (G)
-Polyvinyl alcohol (PVA) and alginate (A)
-Polyhydroxyalkanoates (PHAs)

were tested with different polymers weight compositions and synthetized with different techniques as freeze-drying and salt leaching.
The idea is to recreate a similar environment for the cells (ECM) and so, to obtain a real model of the cancer to better study its behaviour, morphology, characteristics and future targeting therapies.

In addition to these cancer models, one other aim of this thesis was to investigate the transfection efficiency of CRISPR-Cas9 using different non viral vectors.

To achieve our aim, that is the validation of a new safe transfection method to perform the "gene-editing" with the CRISPR- Cas9 system, choosing one of the tumor forms in which an alteration in the expression of the integrin-3 protein has been demonstrated, in vitro and in 3D models transfection assays have been performed using immortalized human tumor cells with lipofectamine.

The idea to functional validate the genes expression modifying PDAC cells using this genetic tool arises from the interest in the possibility to achieve the identication of genes associated with tumor specifics surface proteins that could represent an easiest way to target complex nanoparticles potentially able to selectively attach tumors cells and work as theranostic agent for future therapies.