Barreira hematoencefálica-num-chip: plataforma de deteção neuromimética para estudar a interface nanotecnologia-biologia no tratamento da doença de Alzheimer

NeuroTech4Med aims to go beyond the current state-of-the-art by addressing the two main challenges in neuroscience, particularly in AD: (1) the inefficiency of AD-approved drugs for targeting and bypass the BBB, and (2) the lack of robust BBB-sensing preclinical tools for the screen of drugs to cross the BBB and reach diseased neurological cells. To address these challenges, the project has two main research objectives (RO): RO1: Develop efficiency NFs, using novel nanoengineering methodologies (magnetic and/or fluorescent tracking reports) to cross BBB, target-AD hallmarks and deliver medicines. RO2: Develop a robust BBB model, equipped with a monitoring biosensing system that allows the accurate, automatic and continuous measurement of biomarkers secreted by neuronal biomodels, to determine the efficiency of AD treatment. To accomplish these ROs, four tasks will be developed by a multidisciplinary team that gathers complementary expertise in the fields of nanomedicine, microfluidics, OoC, numerical simulation and micro(bio)sensing (Fig1-2, Annex). Therefore, RO1 will be completed with Task1, under the supervision of the PI, RR, which has experience in develop NFs as drug delivery nanocarriers [20-24]. In Task1, LbNPs, with and without inorganic magnetic core, will be synthesised and fully characterised. This strategy of incorporate magnetic NPs can enable the tracking of the NFs to cross BBB using MRI. However, other NP-tracking strategies will be exploited, such as using fluorescence probes to target AD-hallmarks, as CRANAD-2, creating a NFs-AD tracking model, that to be best of our knowledge, has never been reported (completing RO1). In Task2, led by RR and ST, the neuromimetic device will take-off based on previous technological advances made in the concluded project EXPL/EMD-EMD/0650/2021, where the applicant served as PI. Herein, a BBB model (containing vascular endothelial cells, astrocytes and pericytes) will be combined thought a biomembrane that allows the seeding and spread of the BBB monolayer, creating an extracellular matrix that emulates the in vivo physiological and biochemical microenvironment. To recapitulate the BBB, the brain platform will consist of a multiple-layer chip using a biocompatible, flexible and transparent polymer - PDMS. Different configurations of the microfluidic device will be modelled and pre-validated using simulation software, such as ANSYS, to study parameters such as the transport of diluted species (i.e., diffusion of O2/CO2, nutrients, drugs and biomarkers), velocity fields and mass transport of NFs, coordinated by ST. In Task3, led by GM, the design and fabrication of the multi-biosensing system, will follow porous microneedles integration as sensing aid-tool (developed at EXPL/EMD-EMD/0650/2021,[16]) within the perfusion chamber and connected to standard EC and optical sensors. The biosensing system will be settled using the experience of the team members to computer simulate (VC, ST), develop (PS, RR) and integrate biosensors (GM, PS, JC) in microfluidic devices [10-13,15,25]. In Task4, led by RR, the LbNPs will be used as A?-targeting and drug nanocarrier model to screen the ability to cross the BBB. Task4 will allow the validation of novel NF strategies, gaining potential insight into the mechanism of tracking-NFs to cross BBB and deliver AD-drugs, completing the purpose of this project by delivering a neuromimetic-sensing prototype for the progress of research in AD (completing RO2).