Use of carbon nanotubes for novel approaches towards spinal network repairing

Nanotechnology underwent a very rapid development in the last decades, thanks to the invention of different techniques that allow reaching the nanoscale. The great interest in this area arises from the variety of possible applications in different fields, such as electronics, where the miniaturization of components is a key factor, but also medicine. The creation of smart systems able to carry out a specific task in the body in a controlled way, either in diagnosis or therapy or tissue engineering, is the ultimate goal of a newborn area of research, called nanomedicine. In fact, to reach such an outstanding objective, a nanometer‐sized material is needed and carbon nanotubes (CNTs) are among the most promising candidates. The aim of this thesis was to study this opportunity and, in particular, the possible application of carbon nanotubes for spinal network repairing. After a review of the main features of neuronal network systems and the most common techniques to study their functionality, possible applications of nanotechnology for nanomedicine purposes are considered, focusing the attention on CNTs as neuronal interface in nerve tissue engineering. The work can be divided into two big parts. In the first part the impact of carbon nanotubes on various neuronal systems was studied. Different form of carbonaceous materials (carbon nanotubes, nanohorns and graphene) were deposited in a homogeneous way on a glass surface playing with organic functionalization and different deposition techniques. Hippocampal neuronal cells were grown on their surface to better understand how morphology and conductivity of the material could influence the activity of the neuronal network evidencing how both these characteristics could affect the electrophysiological properties of neurons. Then, also spinal neurons were grown on carbon nanotubes network deposited on a glass substrate to evaluate, for the first time, the impact of carbon nanotubes on this kind of cells. The tight interaction between these two materials appeared to cause a faster maturation of the spinal neurons with respect II to the control grown on a glass substrate. The long-term impact on a complex tissue (spinal cord slice) grown on carbon nanotubes carpet was also studied. The intimate interaction between the two materials observed by TEM and SEM analysis caused an increase in dimensions and number of neuronal fibers that comes out from the body of a spinal cord slice. An increase in electrophysiological activity of all neuronal network of the slice was also reported. In the second part of the work different conductive biocompatible nanocomposite materials based on carbon nanotubes and “artificial” polymers (such as Nafion, PVA, PET, PEI, PDMS and PANI) were investigated. The idea is to test these materials as neuronal prosthesis to repair spinal cord damage. All the prepared scaffolds showed CNTs on the surface favoring CNTs-neurons interaction. To address this aim different techniques and different organic functionalizations of CNTs were utilized to control supramolecular interactions between the nanomaterial and polymers orienting the deposition of the CNTs and preventing their aggregation. After that, an innovative method to study the possible ability of this nanocomposite materials to transmit a neuronal signal between two portions of spinal cord was designed. Functionalization of gold surfaces with thiolated carbon nanotubes have been conducted in order to develop suitable devices for neuronal stimulation and consequent spinal cord lesions repairing. In particular thiol groups were introduced on the graphitic surface of carbon nanotubes by means of covalent functionalization. First of all, the interaction of CNTs with gold nanoparticles has been evaluated, then a gold surface has been coated by means of contact printing technique with a homogeneous film of CNTs. This hybrid material could be useful to produce innovative electrodes for neuronal stimulation