Carbon Nanotube Hydrogels Reveal Threshold‐Dependent Regulation of Neuroblastoma Cell Growth and Maturation by Mechanical and Chemical Factors

Carbon nanotube (CNT)-based hydrogels have the potential to serve as 3D platforms for nerve regeneration. However, the interplay between different cues governing the formation of a complex tissue-like cellular structure is still ambiguous. Herein, two approaches are adopted to develop PVA/CNT hydrogels using phase inversion method and low kinetic gelation, enabling unprecedented CNT loading capacity (75% w/w) without compromising their elasticity. By controlling key factors affecting cell coverage and maturation, including Young's modulus (YM), CNT concentration, and pore size, distinct thresholds are identified where these factors dominate cell coverage. Results demonstrated that when CNT exceeds 60% w/w or a coating is applied to enhance CNT-cell interaction, CNT effect dominates, increasing cell coverage with increasing CNT concentration. However, below a specific YM threshold, YM dominates cell growth, covering up to 50% of the scaffold surface regardless of CNT concentration or exposure. Lastly, controlling the pore size to 100-250 μm further increased cell coverage to >70%, breaking through previous plateau and upregulating TUBB3 maturity marker. Additionally, certain key factors are seen to synergistically codominate in determining cell growth.