Suitability of the Composite Made of Multi Wall Carbon Nanotubes-Polyvinylpyrrolidone for Culturing Invertebrate <i>Helix aspersa</i> Neurons

Carbon nanotubes have been used as scaffolds for tissue engineering. However, the identification of these nanomaterials disperses in biological solutions and their direct interaction with nerve cells is still controversial. The aim of this work is to characterize the topographic and ultra-structural distribution of the composite made of multi wall carbon nanotubes-polyvinylpyrrolidone (MWCNTs-PVP) deposited on the Helix aspersa neurons and over glass coverslip. Scanning Electron Microscopy (SEM) and Confocal Microscopy (CM) studies were done to analyze the properties of such MWCNTs-PVP composite. The cerebral ganglion of Helix aspersa was treated and incubated with MWCNTs-PVP, fixing it in paraformaldehyde at 4% and was observed with SEM and CM. Although the nanotubes were not labeled or stained with fluorescent compounds, the MWCNTs-PVP deposited on glass and on nerve cells, was observed by the confocal microscope in the reflection mode. In SEM studies, it was observed that MWCNTs-PVP was attached to the surface on neurons. Moreover, in CM studies, it was possible to observe that MWCNTs-PVP was attached to the neuronal membrane, crossing the cell membrane and getting into the cytoplasm. These results support the hypothesis that carbon nanotubes interact with the neuronal cell membrane and can be useful for neuronal tissue engineering. In addition, these results open new alternatives for toxicological studies, in order to elucidate the cytotoxicity of MWCNTs-PVP composite in neurons and other excitable cells.