Enhancing Neural Tissue Growth and Maturation Using Polycaprolactone/Polyaniline Conductive Nanofibrous Scaffolds in a Biomimetic Culture System

The growing prevalence of neurological disorders, driven by traumatic brain injuries and neurodegenerative diseases, underscores the urgent need for innovative therapies. Repairing or replacing damaged nerve cells remains challenging due to the limited regenerative capacity of the human nervous system. Traditional 2D cell cultures and in vivo animal models often fall short in replicating the complex cellular environments and interactions found in human tissues. Emerging technologies, including 3D cell cultures, co-culture systems, and bioreactors, offer new opportunities for studying neural cell behavior and mimicking brain functionality more accurately. Additionally, tissue-engineered models provide platforms for developing structured neural cell cultures, advancing the field by creating more physiologically relevant in vitro models that could bridge the gap between preclinical studies and clinical applications. In this project, a cutting-edge culture system was developed to promote neural stem cell growth and maturation in a biomimetic environment. The system is based on an electrospun aligned nanofibrous scaffold obtained from a polycaprolactone (PCL) and polyaniline (PANI) blend, leveraging the conductive properties of PANI to support neural activity. Plasma treatment was applied to the scaffold to enhance its surface properties, facilitating cell adhesion. Neural stem cells (NE- 4C cell line) were then seeded on the scaffold and exposed to a low concentration of retinoic acid (10^(-6) M), which acted as a differentiation agent. Cellular assays demonstrated that NE-4C cells remained viable when cultured on the nanofiber scaffolds, with evidence of neuronal differentiation. Notably, the aligned nanofibers promoted directional neurite extension, following the orientation of the fibers. In addition, NE-4C cells were cultured within IVTech’s LiveBox1 bioreactor in dynamic condition, which offers the capability to apply electrical stimulation to the scaffold. In particular, in vitro tests were conducted to assess the cytocompatibility of the stimulation system performing direct and indirect tests and monitoring cell viability at different time points (24h, 48h, 72h). The obtained results demonstrated that the developed culture system allows for a comprehensive study of neural cell behavior, growth, and interaction within a controlled, biomimetic environment. These initial findings are encouraging for creating advanced platforms for neural regeneration and disease modeling with enhanced functional characteristics. Such platforms represent a valuable approach for understanding disease mechanisms and could aid in the discovery of novel therapies.