Single-layer graphene oxide regulates energy metabolism, enhances protein synthesis and promotes inflammasome complex formation in human platelets

Objectives Graphene-based materials have considerable potential in biomedical applications; however, their effects on platelet metabolism and function remain incompletely understood. This study investigated the effects of graphene oxide (GO) on platelet bioenergetics, de novo protein synthesis, NLRP3 inflammasome activation and prothrombotic responses. Methods Human platelets were exposed to GO to assess glucose uptake, lactate production, mitochondrial reactive oxygen species, protein synthesis, NLRP3 inflammasome activation and procoagulant responses using biochemical, flow cytometric, immunofluorescence and functional assays. The involvement of glycolysis, mTOR signaling and NLRP3 inflammasome activation was examined using dichloroacetate, rapamycin and MCC950, respectively. GO-induced thromboembolic injury was evaluated in vivo in mice using multimodal ultrasound/photoacoustic imaging and histopathological analysis. Results GO exposure induced a Warburg-like metabolic shift characterized by increased glucose uptake, lactate production and GLUT3 surface expression. GO also stimulated de novo synthesis of pro-IL-1β, NLRP3 and tissue factor in a puromycin-sensitive manner. Inflammasome activation was supported by increased caspase-1 activity and NLRP3-ASC co-localization. Pharmacological inhibition of glycolysis and mTOR signaling attenuated GO-induced metabolic and inflammasome responses. Furthermore, GO promoted a procoagulant phenotype, evidenced by increased phosphatidylserine exposure and accelerated clot formation, which was partially attenuated by MCC950. In vivo , intravenous GO administration induced pulmonary thromboembolism accompanied by reduced pulmonary vascular perfusion and oxygen saturation. Conclusions GO induces coordinated metabolic, inflammatory and procoagulant changes in human platelets, characterized by enhanced glycolytic activity, mTOR-associated signaling, NLRP3 inflammasome activation and procoagulant responses. These findings provide mechanistic insight into GO-induced platelet dysfunction and emphasize the importance of evaluating hemocompatibility and thromboinflammatory effects when developing graphene-based materials for biomedical applications.