Overall the project, the consortium has developed and introduced novel technologies to improve cardiac and neuronal in-vitro toxicology. These technologies stem from the original TOX-Free ideas but expanded into different and novel applications that followed the general needs and trends of the toxicology field for neurons and cardiomyocytes.
Regarding cardiotoxicity, the consortium has developed a new device concept for the measurement of cardiac contraction forces with high spatial resolution, down to single cell, in large monolayer cultures. In addition, these measurements have been combined also with electrical recording of field potentials, to move toward multiparametric analyses. Additionally, the consortium has developed protocols for extreme long-time Action Potential recordings from cardiomyocytes on microelectrode arrays (MEA) for enabling new insights into chronic cardiotoxicity. Finally, we demonstrate a novel polymer based transistor that enables the recording of intracellular-like cardiac Action Potentials by exploiting laser cell optoporation. These polymer based transistors can potentially be fabricated and produced with very low-cost techniques and can be more environmental friendly than semiconductor based biosensors. Furthermore, the consortium has established new protocols for assessing cardiotoxicity on MEA by comparing the effects of several compounds on different cardiac cell models derived from human stem cells, determining which model can better resemble the human cardiac model for cardiotoxicity. Finally, we have also presented a new method for measuring in-vitro fiber-like structures of cardiomyocytes that can reveal quantitative information on signal conduction velocity and gap junctions.
Regarding neurotoxicity, we have developed new all-optical technologies based on Raman Spectroscopy for cell phenotyping and for assessing neuronal toxicity at earlier stages than established technologies. These optical technologies enabled us to discriminate differentiation stages of neuronal spheroids with high temporal resolution and enabled us to detect toxic effects of known compounds on neurons at earlier stages without damaging the spheroids. Potentially, these optical technologies can offer the possibility to follow the development of neuronal toxicity over time in complex three-dimensional spheroids or organoids. Moreover, we have also shown that Raman Spectroscopy combined with MEA electrophysiology can reveal subtle toxicological effects in neuronal cultures by correlating the optical and electrical measurements with advanced machine learning algorithms.
Finally, we have also introduced a laser cell poration technology that offers the recording of sub-threshold activity in neuronal culture on MEA in a non-invasive manner, enabling longitudinal analyses.
The consortium has been active in disseminating and communicating these results to the scientific community and the general audience. Among the main activities, the consortium has actively organized and supported the first three editions of the Biocube Meeting, a new winter-school and workshop focused on biophotonics, bioelectronics, and biomechanics.
