Electrochemically optimized multi-component polyacrylonitrile nanofiber scaffolds as a platform for three-dimensional glioblastoma cell culture

Glioblastoma multiforme represents the most aggressive primary brain malignancy with limited treatment options. Current research critically lacks three-dimensional culture platforms that can both recapitulate native tumor microenvironments and enable quantitative monitoring of cellular behavior. This study presents the first electrochemically monitored three-dimensional glioblastoma culture platform developed using multi-component PAN nanofiber scaffolds. Six sensor configurations incorporating Ho-MOF, C500, and GO were systematically developed: ITO/PAN, ITO/PAN/MOF, ITO/PAN/C500, ITO/PAN/GO, ITO/PAN/GO/MOF, and ITO/PAN/GO/C500. Electrochemical impedance spectroscopy unveiled a critical discovery: optimal electrical performance does not guarantee biological compatibility. GO/MOF exhibited lowest electron transfer resistance (31.3 Ω) yet failed completely in cell adhesion. C500-modified scaffolds achieved the important finding 39.8 Ω resistance with > 95% cell viability. Cyclic voltammetry delivered current responses (280.53 μA). Precisely-controlled electrospun nanofibers (392-507 nm diameters, 9-10 μm pores) enabled authentic three-dimensional cellular infiltration. Cytotoxicity testing and Hoechst staining of U-87 MG and LN-18 glioblastoma cells demonstrated high cell viability and a well-organized cellular distribution, particularly on C500-modified PAN nanofiber scaffolds among the tested compositions, consistent with three-dimensional culture conditions. This platform obliterates the gap between reductionist 2D models and clinical reality, delivering real-time electrochemical monitoring within physiologically-relevant three-dimensional scaffolds for transformative glioblastoma research applications.