Final Report Summary - MESOTAS (Chatting with Neurons: A novel approach to the study of neurophysiologic responses of neuronal tissue in vitro, combining nanotechnology, tissue engineering, microfluidics and neuroelectrophysiology)
The MESOTAS research team developed a number of new nanotechnologies to be able to investigate neuronal cells and their ability to form neuronal networks. These novel approaches study the neurophysiologic responses of neuronal tissue in vitro, combining nanotechnology, tissue engineering, microfluidics and neuroelectrophysiology. One of the main research devices is a micro- and nanofabricated multi-electrode array (MEA) containing 3D-pores. The MESOTAS technology platforms have gained immense interest in neuroscience and -technology because of improved spatial resolution and ease of handling in 3D cultured neurons. More recently, Laboratory on a Chip technology was introduced in the field of neuroscience. We contributed to this field of work by reducing the complexity of animal models by brain on chip technology and by reducing the number of animals required in pre-clinical research.
Here, the combination of microfluidics, tissue engineering and neuroelectrophysiology on advanced MEA-chip technology platform has been realized. Because neuronal tissue on chip differs from the neurons in their natural environment, the first objective was to follow a systems engineering approach to realize a platform technology, which allows us to reliably co-culture cells in a 3D interconnected configuration and analyze their morphology. We realized such culture system by the out-of-plane integration of microfluidic device, a microbioreactor, which provides an artificially vascularized system to the MEA chip. For on-line monitoring of the culturing conditions, we implemented also a micro-total analysis systems (TAS) with the microbioreactor. Being able to retrieve a sample from the microbioreactor culture chamber utilizing microchip capillary electrophoresis, potentially we are able to couple such an analysis device with mass spectrometry. Such an approach paves the way to correlate electrophysiology with neurochemistry. Although such technology validation is still ongoing, we have put this approach into context of the different length scales of biology and realized a miniaturized culture chamber with integrated active microfluidic and nano-functionality. Previously, it has been demonstrated that physical and chemical micro- and nanostructures influence cell guidance, viability and cell differentiation. So far, unfortunately without a unifying theory to explain the involved mechanisms. Therefore, our second objective was to further our understanding with respect to the influence of nanocues, implementing microfluidic programming to activate porous nanostructures in the MESOTAS 3D culture approach. We were able to investigate cellular signaling and pathway reactions in relation to the cell’s adhesion mechanism within the porous nanostructure of the scaffolds. Combining the first (3D culture) and the second objective (active porous nanostructures) will allow us to work towards clinical questions of neurodynamic diseases such as epilepsy.
Of course, there is ample of room for improvements of the MESOTAS technology platforms, however, the systems achieved so far point us towards a confirmation of our hypothesis that for neuronal disorders, 3D cell co-culture models resemble the natural neural networks more closely than 2D and show a larger extent of neuronal activity in the astrocytes of the neuronal network. Subsequently, our new 3D MESOTAS culture platforms serve as a novel brain model to study therapeutic procedures, for instance, by selective neurostimulation using our newly developed nanomechanical actuator chip. Demonstrating the possibility of local microfluidically programmed actuation allowed us to confirm our third objective that nanostimulation of neuronal subsystem within our 3D-MESOTAS cell culture model can be utilized for the in vitro investigation of neuroelectrophysiologic dynamics of brain diseases.
Here, the combination of microfluidics, tissue engineering and neuroelectrophysiology on advanced MEA-chip technology platform has been realized. Because neuronal tissue on chip differs from the neurons in their natural environment, the first objective was to follow a systems engineering approach to realize a platform technology, which allows us to reliably co-culture cells in a 3D interconnected configuration and analyze their morphology. We realized such culture system by the out-of-plane integration of microfluidic device, a microbioreactor, which provides an artificially vascularized system to the MEA chip. For on-line monitoring of the culturing conditions, we implemented also a micro-total analysis systems (TAS) with the microbioreactor. Being able to retrieve a sample from the microbioreactor culture chamber utilizing microchip capillary electrophoresis, potentially we are able to couple such an analysis device with mass spectrometry. Such an approach paves the way to correlate electrophysiology with neurochemistry. Although such technology validation is still ongoing, we have put this approach into context of the different length scales of biology and realized a miniaturized culture chamber with integrated active microfluidic and nano-functionality. Previously, it has been demonstrated that physical and chemical micro- and nanostructures influence cell guidance, viability and cell differentiation. So far, unfortunately without a unifying theory to explain the involved mechanisms. Therefore, our second objective was to further our understanding with respect to the influence of nanocues, implementing microfluidic programming to activate porous nanostructures in the MESOTAS 3D culture approach. We were able to investigate cellular signaling and pathway reactions in relation to the cell’s adhesion mechanism within the porous nanostructure of the scaffolds. Combining the first (3D culture) and the second objective (active porous nanostructures) will allow us to work towards clinical questions of neurodynamic diseases such as epilepsy.
Of course, there is ample of room for improvements of the MESOTAS technology platforms, however, the systems achieved so far point us towards a confirmation of our hypothesis that for neuronal disorders, 3D cell co-culture models resemble the natural neural networks more closely than 2D and show a larger extent of neuronal activity in the astrocytes of the neuronal network. Subsequently, our new 3D MESOTAS culture platforms serve as a novel brain model to study therapeutic procedures, for instance, by selective neurostimulation using our newly developed nanomechanical actuator chip. Demonstrating the possibility of local microfluidically programmed actuation allowed us to confirm our third objective that nanostimulation of neuronal subsystem within our 3D-MESOTAS cell culture model can be utilized for the in vitro investigation of neuroelectrophysiologic dynamics of brain diseases.