Exploring the Potential of Aligned Electroconductive Nanofibrous Membranes for Culturing Peripheral Nervous System Cells

A large variety of substances (i.e. pollutants and therapeutics) have been reported to cause peripheral demyelinating neuropathies that result from the injury to Schwann cells and sensory/motor neurons, by direct toxic effect or immune-mediated cytotoxicity. In vitro myelination/demyelination assays are carried out using organotypic cultures of dorsal root ganglia rat explants, so far. In this work, we proposed the development of an in vitro model reassembling the key features of the peripheral nerve composition by co-culturing Schwann cells (SC) and neurons on an engineered substrate having instructive cues to steer cell assembling. Instructive substrates were produced by electrospinning technology using a blend of polycaprolactone (PCL) and polyaniline (PANI) doped with Camphor-10-sulfonic acid (CSA). PCL, chosen as the core polymer of fibers, is characterized by high cytocompatibility and processability, whereas, PANI doped with CSA was added to enhance the conductive properties of the mats. Random or aligned oriented fiber were produced and characterized in terms of physicochemical (hydrophilicity, stiffness, electroconductivity and composition) and structural (topography, alignment and diameter of the fibers). Finally, preliminary in vitro experiments were carried out by seeding rat-derived SC onto our membranes to assess cell viability and proliferation by CellTiter-Blue® assay and cell organization by DAPI/Phalloidin staining. Thanks to these preliminary tests, we can state that PCL-PANI electrospun membranes represent a promising system for culturing peripheral nervous system cells. In particular, the aligned mats helped the cells to have a more physiological morphology and organization, replicating the anisotropic structure of in vivo nervous tissue. Acknowledgment: This project has received funding from Humatoxchip project - Bando Trapezio, Compagnia Sanpaolo and “Bando per il finanziamento ex-post di progetti di ricerca di Ateneo”, Compagnia San Paolo.