Carbon-based nanomaterials induce ovarian dysfunction via NLRP3 inflammasome-mediated pyroptosis in granulosa cells

Carbon-based nanomaterials have attracted increasing attention due to their routine exposure and potential health risks. However, the impact of carbon-based nanomaterials on ovarian function remains poorly understood.

This study aimed to systematically evaluate the ovarian toxicity of three representative carbon-based nanomaterials and to elucidate the underlying mechanisms of ovarian dysfunction.

Female mice were exposed to multi-walled carbon nanotubes (MWCNTs), graphene, or fullerene. Estrous cycle, hormone levels, and follicular development were assessed. Transcriptomic sequencing and molecular analyses were performed to explore potential mechanisms, and the protective effect of NLRP3 inhibition was validated using mouse, granulosa cell, and human ovarian cortex models.

All three carbon-based nanomaterials induced estrous cycle disruption in mice. Hormone analysis revealed elevated serum follicle-stimulating hormone levels in all exposure groups, while a significant decrease in estradiol was observed only in MWCNTs group. MWCNTs and graphene significantly reduced the numbers of primordial and growing follicles and increased the number of atretic follicles. In contrast, the fullerene exhibited milder effects, with only an increase in atretic follicles. None of the three nanoparticles significantly affected female fertility in mice. Further transcriptomic analysis identified the NLRP3 inflammasome as a key mediator of granulosa cell injury. Unlike classical apoptotic pathways that mediate granulosa cell death, exposure to carbon-based nanomaterials induced pyroptosis in granulosa cells by activating the NLRP3 inflammasome pathway, subsequently leading to follicular atresia. Notably, intervention experiments using granulosa cells, mouse model, and cultured human ovarian cortex demonstrated that the NLRP3 inhibitor MCC950 effectively alleviated carbon-based nanomaterials induced ovarian dysfunction.

These findings reveal a novel mechanism by which carbon-based nanomaterials impair ovarian function through NLRP3 inflammasome-mediated pyroptosis and highlight NLRP3 inhibition as a potential therapeutic strategy for mitigating carbon-based nanomaterials associated ovarian injury.