| dc.description.abstract | The dynamics of the nervous system are characterized by the intricate interactions
among precisely arranged multiscale neural networks, arising from the innate capacity
of the system for self-organization and adaptation to regulatory cues from the microenvironment.
However, the fundamental principles governing organization and function of neural networks remain
largely unknown. In recent years, in vitro engineered interfaces have emerged as a viable approach
to study neural networks at the mesoscale, providing a highly controlled microenvironment and
allowing to mimic key aspects of in vivo neural networks, while minimizing the associated complexities
by adopting a reductionist approach. In this M.Sc thesis, we used novel models for in vitro neural
culturing to recapitulate significant aspects from in vivo topology and manipulate the structure-function
dynamics within populations of neurons. To attain modular organization of segregated neuronal
populations we used novel multi-nodal microfluidic devices, featuring afferent-efferent connectivity
promoted via Tesla-valve inspired microchannels. The presence of structural modularity and unidirectionality
was validated by employing viral tools for cellular delivery of genes for expressing distinct fluorescent
proteins within the segregated neural population in two-nodal microfluidics. To capture their functional
dynamics, we implemented and optimized a system for optogenetics and calcium imaging.
Furthermore, we demonstrate that altering the number of microchannels within a five-nodal microfluidic
coupled with microelectrode array can influence inter-nodal connectivity. We then selectively perturbed
one node within the five-nodal microfluidic and showed notable alternations in functional connectivity
among the nodes, as well as in firing rates within each node. Finally, we tested the biocompatibility of
novel 3D microfabricated interfaces with neuronal cultures, aiming to facilitate the formation of tridimensional
neural networks while retaining the simplicity of handling neural cultures. This work highlights the immense potential of advanced neuroengineered in vitro models to recapitulate microscale topological organization
of neural networks and investigate their structure-function dynamics in healthy and perturbed conditions. | |