Mariana Seke1, Ivan Jovanović2, Nadja Trklja2*, Nataša Mačak Stefanović2, Jovana Kuveljić2, Maja Živković2 and Aleksandra Stanković2
1Institute of Nuclear Sciences ''Vinca'', University of Belgrade
2''Vinca'' Institute of Nuclear Sciences, University of Belgrade
nadja.trklja [at] vin.bg.ac.rs
Abstract
Fullerenols C60(OH)n are widely regarded as biocompatible nanomaterials at low concentrations, but higher doses may induce cytotoxic effects. Previous studies indicate mitochondrial damage as a mechanism of toxicity, yet the broader molecular pathways involved remain poorly characterized.
We performed a transcriptomic reanalysis of publicly available dataset GSE3364 to investigate gene expression changes in human umbilical vein endothelial cells treated with fullerenol C60(OH)24 (100 μg/ml, 24 h). Differentially expressed genes (DEGs) were identified using a linear model with empirical Bayes statistics in the limma R package. Functional pathway and biological process enrichment analyses were conducted with iPathwayGuide (Advaita Bio), using thresholds of |log2 fold change| ≥ 0.6 and p ≤ 0.05.
Analysis identified 2161 DEGs, revealing enrichment of pathways related to membrane-associated processes. Notably, cell adhesion molecules, neuroactive ligand–receptor interaction, and other membrane-linked signaling pathways were affected. These findings suggest that fullerenol-induced cytotoxicity may involve perturbations of membrane integrity, receptor-mediated signaling, and cellular adhesion, complementing previous observations of mitochondrial dysfunction. Additional pathways, including those related to extracellular matrix organization, were also implicated, indicating a multi-faceted cellular response to high-dose fullerenol exposure.
Our results highlight membrane-associated pathways as potential mediators of fullerenol C60(OH)24 cytotoxicity in endothelial cells. Understanding these mechanisms is crucial for optimizing biomedical applications, including dosing strategies, treatment duration, and administration routes. This study provides new insight into the molecular effects of fullerenols and supports further research into their safe use in nanomedicine.
Keywords: fullerenol, transcriptomics, DEGs, pathways, HUVECs,
Acknowledgement: This research was supported by the Serbian Ministry of Science, Technological Development, and Innovation, Grant No. 451-03-33/2026-03/ 200017