SCHWANN CELLS BEHAVIOR ON SMART ELECTROCONDUCTIVE BIOMATERIALS FOR PERIPHERAL NERVE REGENERATION

Paravizzini G^1,2, Sandu E¹, Fregnan F^1,2, Shirosaki Y³ and Raimondo S^1,2 | ¹Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, Turin, Italy; ²Cavalieri Ottolenghi Neuroscience Institute, University of Turin, Regione Gonzole 10, Orbassano, Turin, Italy; ³Faculty of Engineering, Kyushu Institute of Technology, 1-1 Sensui-cho, Tobata-ku, Kitakyushu-shi, Fukuoka, Japan

Functional recovery following peripheral nervous system (PNS) injuries remains a significant challenge for clinical research. The PNS exhibits intrinsic regenerative capacity in which Schwann cells play a pivotal role, with their remarkable phenotypic plasticity, capable of modulating several processes to achieve axonal regeneration. Despite this intrinsic potential, when the injury is severe, with a gap exceeding 5 mm, or the surgical repair is delayed, recovery is often unsatisfactory, resulting in the formation of a non-functional nerve. The current clinical gold standard is autograft, however, it is associated with notable limitations, such as the sacrifice of healthy nerve tissue and potential surgical complication. Among the strategies aimed at promoting functional PNS regeneration, biomaterials have emerged as a promising alternative, thanks to their ability to provide mechanical support and recreate an extracellular environment that promotes regenerative responses. In the present study biocompatible and biomimetic biomaterials based of chitosan/carboxymethylcellulose were developed (Ch70CMC30 scaffolds). Reduced graphene oxide (RGO), an electroconductive biomaterial, was incorporated into these starting biomaterials, both in bulk form at different concentrations (0.3%, 0.6%, 1.0%) and as surface striped micropatterns with two distinct dimensions (100 μm and 200 μm). The glial cell behaviour to the films was assessed in terms of proliferation, morphological responce and protein expression. In the initial experimental phase, the RT4-D6P2T glial cell line derived, from Schwannoma, was used to conduct the experiments. Morphological analysis was then also performed with primary culture of rat Schwann cells, to evaluate a model more representative of the regenerative environment. The scaffolds demonstrated efficacy in supporting consistent cell growth and stable expression of markers associated with cell survival and proliferation, confirming their cytocompatibility. In particular, in the case of RGO micropatterns, morphological analysis revealed preferential cell adhesion to RGO coated regions and a pronounced alignment along the pattern. These findings highlight the potential of RGO-functionalized scaffolds in modulating cell growth and migration, laying the foundation for future studies pointing at developing advanced regenerative strategies.