Periodic Reporting for period 2 - BrainChip4MED (Brain-on-a-chip as a preclinical model tool for the screening of theranostic nanoformulations for neurodegenerative diseases)
Reporting period: 2023-03-01 to 2024-02-29
Dementia, which includes a broad category of brain disorders that decline brain function, has for a long time been a global challenge that was dismissed by pharmaceutical companies due to the low penetration efficiency of drugs that are able to cross the blood-brain barrier (BBB), as well as to the limited understanding of the underlying biophysical mechanism, which is still mostly unknown.
Recently, brain-on-a-chip (BoC) has emerged as an advanced microfluidic platform combined with 3D tissue culture techniques, with the potential to create an accurate and simple-to-use preclinical model tool, by decoupling a complex organ, such as the brain, into different cellular structures, while maintaining their interconnections. Also, the possibility of integrating biosensors on it could extend its monitoring and workability for longer periods of time.
Motivated by the lack of an appropriate in-vitro model to study brain-targeting drug nanocarriers, and the potentiality of BoC technology to recapitulate human biology and predict in-vivo response, the overreaching aim of this action is to develop an advanced microfluidic preclinical device with the structural and functional aspects of the brain tissue and BBB, using BoC as a preclinical model tool to assess and study possible alternatives for the diagnosis and treatment (i.e. theranostic) of neurodegenerative disorders, and to develop multifunctional novel stimuli-responsive drug nanocarrier systems (Figure 1).
By so, BrainChip4MED prototype (mimicking the neural tissue and BBB integrated with multiplexed biosensors) proposed in this engineered platform is a cutting-edge technology that allows for the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease (AD).
Overall, the most innovative achievements during this action were (1) the development and optimisation of a novel pro-angiogenic hydrogel for (bio)engineering of the BBB and, (2) the development of a novel strategy of optical biosensing of AD hallmarks (i.e. amyloid beta) for the continuous and automatised monitoring of the AD models during preclinical trials. These scientific and technological achievements create an innovation in the current state-of-the-art for BoC as preclinical tools. The presented technology, represent a first step to achieve automatised BoCs with high potential to impact the society by delivering an advanced microfluidic device capable to reduce the use of animal tests, increase the successful rates in the translation of novel nanoformulations from laboratory to clinical use, and find a successful strategy that allows a better understanding and treatment of AD and dementia.
Recently, brain-on-a-chip (BoC) has emerged as an advanced microfluidic platform combined with 3D tissue culture techniques, with the potential to create an accurate and simple-to-use preclinical model tool, by decoupling a complex organ, such as the brain, into different cellular structures, while maintaining their interconnections. Also, the possibility of integrating biosensors on it could extend its monitoring and workability for longer periods of time.
Motivated by the lack of an appropriate in-vitro model to study brain-targeting drug nanocarriers, and the potentiality of BoC technology to recapitulate human biology and predict in-vivo response, the overreaching aim of this action is to develop an advanced microfluidic preclinical device with the structural and functional aspects of the brain tissue and BBB, using BoC as a preclinical model tool to assess and study possible alternatives for the diagnosis and treatment (i.e. theranostic) of neurodegenerative disorders, and to develop multifunctional novel stimuli-responsive drug nanocarrier systems (Figure 1).
By so, BrainChip4MED prototype (mimicking the neural tissue and BBB integrated with multiplexed biosensors) proposed in this engineered platform is a cutting-edge technology that allows for the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease (AD).
Overall, the most innovative achievements during this action were (1) the development and optimisation of a novel pro-angiogenic hydrogel for (bio)engineering of the BBB and, (2) the development of a novel strategy of optical biosensing of AD hallmarks (i.e. amyloid beta) for the continuous and automatised monitoring of the AD models during preclinical trials. These scientific and technological achievements create an innovation in the current state-of-the-art for BoC as preclinical tools. The presented technology, represent a first step to achieve automatised BoCs with high potential to impact the society by delivering an advanced microfluidic device capable to reduce the use of animal tests, increase the successful rates in the translation of novel nanoformulations from laboratory to clinical use, and find a successful strategy that allows a better understanding and treatment of AD and dementia.
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.
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.
The expected outcomes and impacts of this research project are relevant at technological, human health and economic levels, and are fully aligned with the UN Sustainable Development Goal (SDG 3) – Good health and well-being. Specifically, the analytical BoC tool and NFs for AD, in development in this action, are essential to understand the progress and prognosis of neurodegenerative diseases, and to support therapeutic decisions of drug therapy (currently ineffective). The BoC prototype is expected to deliver an innovative advanced preclinical platform that mimics the human brain physiology and surpass the use of animal models (3R’s), which besides the ethical issues, is the main responsible for the bias results and failure of new medicines in clinical trials. Thus, this project delivers for the first time, a prototype that integrates in the same platform, BBB and brain models in situ monitored by biosensor’s analysis. This achievement represents a step further in the automatisation and robustness of these advanced preclinical platforms, with the potential to reduce the typical extended costs and time consumed in the development/screening of new medicines. Thus, possibly instigating the research of new strategies to treat dementia at lower market prices. Also, nanomedicines in study show the potential to increase the efficiency of drugs for AD, decreasing their side effects and ensuring better population ageing condition. Moreover, the analytical tool/prototype obtained in this action has great potential as a commercial product. At the same time, the proposed application has great outreach for academia and pharmaceutical research, contributing to the advancement of nanomedicine and neurosciences in Europe and beyond.