Three-dimensional cytosensor for glioblastoma cell analysis using coumarin-integrated nanofiber scaffolds on ITO electrodes

Glioblastoma is the most common and aggressive brain tumor. In this study, a three-dimensional (3D) platform was developed for glioblastoma cell growth and electrochemical analysis. Polyacrylonitrile (PAN) nanofiber scaffolds, serving as a 3D support for U-87 MG and LN-18 cell proliferation, were deposited onto indium tin oxide (ITO) surfaces, enabling electrochemical measurements. The scaffolds were further functionalized with fibronectin and collagen IV. Cellular morphology was assessed by fluorescence microscopy, and electrochemical measurements were conducted to evaluate the analytical performance of the platform. Cell distribution and structural organization were characterized by fluorescence imaging and hematoxylin & eosin (H&E) staining. The electrochemical responses of cells on nano-fiber coated ITO surfaces were determined. Coumarin (C500)-integrated PAN nanofiber scaffolds significantly enhanced the viability and morphological organization of glioblastoma cells, with further improvement observed in U-87 MG cells upon ECM functionalization, particularly with collagen IV. For the ITO/PAN/C500/FN1/LN-18 cytosensor, the limit of detection (LOD) was determined to be 12 cells/mL, the limit of quantification (LOQ) 30 cells/mL, and the relative standard deviation (RSD) 0.34 %. For the ITO/PAN/C500/COL10/U-87 MG sensor, the LOD was 60 cells/mL, LOQ 200 cells/mL, and RSD 1.05 %. This 3D platform, designed to support glioblastoma cell growth, could provide a robust in vitro basis for anticancer drug screening and preclinical investigations, and with further development it may be adapted to evaluate chemotherapy response using patient-derived tumor samples.