The main scientific and technological achievement of WP1 was the biofabrication of the BoC platform comprising both brain and BBB models. With this aim, a novel soft-hydrogel was designed to mimic the BBB ECM and act as a bio-membrane that separates the vascularized cells from the brain. This BBB model consisted of a gelatine-based hydrogel incorporating heparin, as a pro-angiogenic growth factor immobilization molecule, and hyaluronic acid (HA) as an enhancer of the mechanical strength and regulator of the physiological processes related to BBB cells. The hydrogels were fully characterised in terms of mechanical properties, swelling, biodegradation, cell adhesion and proliferation, as well as gene expression. The results shown that the innovative bio-membrane is able to sustain and generate ideal conditions for the spread, adhesion and close junction of endothelial cells for several days, which represents a major advance in the current state-of-the-art to develop mimicking BBB models. This work presented in a Poster communication at the 2022 Discovery Brigham (Boston, USA).
In the return phase, WP2 was initiated with the synthesis of solid lipid nanoparticles (SLNs) described in literature as potential drug delivery nanosystems for the treatment of several neurodegenerative diseases. Several synthesis routes were tested to obtain optimised SLNs using bee wax as lipid precursor. Overall, our optimised synthesis route was achieved by ultrasonication mixing bee wax, tween 80 (as surfactant), PEG-DSPE-NH2 (as surface link), Resveratrol (a neuroprotective compound that increase the inhibition on amyloid beta peptide) and finally functionalised with transferrin (as a molecule to aid active transport across the BBB). In parallel, WP3 was initiated aimed to screen the developed BBB-targeting drug SLNs using BoC as a preclinical tool for the AD’s treatment. The validation of the BrainChip4MED prototype in dynamic flow was conducted during at least 5 days, in continuous flow. Additionally, immunostaining and live/dead analysis techniques were performed to screen the viability of the biomodels along the dynamic in-vitro culturing, and the BoC prototype used to screen the SLNs developed to cross BBB and deliver therapeutic compounds to the brain model. Also, the development of an optical biosensing system to detect AD's hallmarks was tracked, adding innovation to this action. Some of these results were presented as oral communication (Podium presentation) in the 33rd Annual Conference of the European Society for Biomaterials (ESB2023), Davos (Switzerland). The Researcher has also participated in the Fair INL Open Day 2024, presented a poster communication at INL (Annual Symposium), and served as organisation member of the workshop: Nanotechnology – the future of medicine, at XVI Minho Medical Meeting, Portugal.
