Bioengineered in vitro models of human tissues and drug design and development.

This work is developed around two macro-themes, with a common purpose: the development of new drugs to fight inflammatory states that affect the retina and cardiovascular system. The dysregulation of metalloproteinases enzymatic activity is a major feature in many severe pathologies. The creation of an in vitro model of the sub-retinal space, the study of the metalloproteinase families’ regulation and their proteolytic activity, can provide key tools and insights for the design of suitable inhibitors of metalloproteinases, capable of counteracting the dysregulation of these enzymes involved in the progression of inflammatory diseases. The two topics pursued are: 1. Fabrication and characterization of a biotechnological Bruch’s membrane(BM) for an in vitro model of the Human Outer Blood-Retinal Barrier (oBRB). The aim of this work is to fabricate a biotechnological in vitro model BM to test the permeability and safety of novel drug candidates and to study their impact on retinal cells in physiological and AMD conditions. The results have proven that electrospun mats are capable to reproduce the in vivo properties of BM, indicating that these membranes can be used as a substrate for retinal cells cultivation. 2. Investigation of versicanase activity of ADAMTS (A Disintegrin And Metalloproteinase with Thrombospondin motifs) proteases. This project aimed to set-up and optimize a western blot-based versicanase assay that can be used to screen novel glycosylated compounds identifying ADAMTS variant-specific inhibitors. The study of new selective exosite ADAMTS-4 and -5 inhibitors showed good preliminary results, encouraging further studies on glycoconjugate compounds, as potential therapeutic agents for inflammatory processes.