Overall strategy

Figure illustrates in a diagram how these three aims will be achieved and the interplay between the different components of the proposal. We will fabricate surfaces with specific nanopatterns with a variety of geometries and substrates (WP1). These nanostructures will also be printed at the nanoscale with specific patterns of molecules (WP2). These surfaces will be integrated on a conventional MEA and a perfusion system (WP3) in order to allow electrical recordings and to perform appropriate changes of the extracellular medium where cells are cultivated. The effect of nanopatterns on cell growth will be tested in two configurations: in the Neuro-Flip-Chip model (see Fig.6) where the nanopatterns will be lowered onto the neurons from above and by growing stem cells and neurons over a nanopatterned surface.

The methodology for exploring the interaction between nanostructures and cells will be based on several experimental tools able to obtain a nanoscale resolution. Some of these methodologies are already available to members of the present consortium, such as AFM and quantum dots and confocal microscopy. Other techniques require technological development to be carried out within the present proposal, such as electrochemical probes and sensors (WP4) and Electron Microscopy and Tomography (WP5).

Micro and nanopatterned substrates will be used to explore and understand the interactions of these nanostructures with stem cells, neurons and neuronal networks (WP6, 7 and 8). WP9 will establish methods in a statistical and significant way: which biological effects nanopattern and nanoprints will have on cells and neurons. These devices will be coupled with conventional optical tweezers for a controlled physical and chemical manipulation at a micro and nanoscale (WP10). WP11 will be dedicated to the evaluation of health risks of the new technologies under development in Nanotechnology, to the management of the project and to the dissemination of the obtained results. Nanopatterns will be produced primarily by CNR-INFM and NMI, while ENS and DTU will provide their expertise and access to their own facilities when necessary. ENS, SISSA and DTU will analyse the biological impact of different nanopatterns. GVT will support SISSA in the preparation of biological samples and provide some software tools for imaging/data analysis. MCS and NMI will integrate these nanopatterns to MEAs and will fabricate an appropriate micro/nanofluidic system.