Uncovering the potential mechanisms of aspartame-induced oral leukoplakia via integrated network toxicology and molecular dynamics simulations

Objective: Aspartame, a widely used artificial sweetener, has attracted increasing attention for its potential systemic and oral toxicity. However, the molecular mechanisms by which it induces oral diseases remain largely unexplored. This study aims to investigate the potential molecular mechanisms of oral leukoplakia (OLK) induced by aspartame.

Methods: This study conducted an integrative analysis of multiple gene expression datasets associated with OLK, combining network toxicology, Mendelian randomization, machine learning, and molecular dynamics simulations to investigate the interactions between aspartame and disease-related target genes. Subsequently, preliminary validation was conducted using clinical OLK tissues, cell lines, and C57BL/6 mice, respectively.

Results: A total of 27 candidate genes were identified as potential molecular targets linking aspartame exposure to the pathogenesis of OLK. Among them, ITGAV, PLAU, MMP16, and NOS3 were recognized as core genes with the highest diagnostic value. These genes were significantly upregulated in OLK tissues and represented key regulatory nodes associated with extracellular matrix remodeling, oxidative stress, angiogenesis, and epithelial dysplasia. Molecular simulations suggested potential interactions between aspartame and these target genes. Both in vitro and in vivo experiments further confirmed that aspartame exposure markedly upregulated the expression of these genes and was accompanied by histopathological alterations indicative of epithelial dysplasia.

Conclusion: This study presents an exploratory, integrative analysis identifying molecular features potentially linking aspartame exposure with OLK-associated epithelial phenotypes. The findings support the generation of testable hypotheses regarding ITGAV-, PLAU-, MMP16-, and NOS3-related pathways but do not establish exposure-response relationships or causal mechanisms. Further studies incorporating physiologically relevant dosing, pharmacokinetic validation, appropriate metabolic controls, and pathway-specific interventions are required to determine the biological relevance of these observations to real-world oral exposure.