Developmental vulnerability to fluoride toxicity: enamel and clearance differences in adolescent versus mature mice

Fluoride plays a dual role in dental health—preventing caries at optimal levels but causing fluorosis when excessive. While most animal studies focus on young mice, age-related susceptibility to fluoride remains poorly understood. This study presents the first comprehensive analysis of developmental stage–dependent differences in fluoride toxicity, focusing on enamel formation and systemic fluoride clearance. Male C57BL/6J mice—adolescent (5–9 weeks) and mature (16–20 weeks)—were exposed to fluoride in drinking water (0, 50, 100, or 125 ppm) for 6 weeks. Adolescent mice developed pronounced dental fluorosis, characterized by chalky white incisors, elevated Quantitative Light-induced Fluorescence (QLF) values, reduced enamel microhardness, and lower enamel mineral density (EMD). Histological analysis revealed disrupted ameloblast morphology, reduced KLK4 expression, and aprismatic enamel, with more severe effects in adolescents. In contrast, mature mice exhibited minimal changes in QLF, enamel hardness, and EMD. Systemic fluoride analysis showed significantly lower serum and urinary fluoride levels in adolescent mice compared to mature mice, indicating reduced excretion and increased tissue accumulation. These findings demonstrate that younger mice are more vulnerable to fluoride-induced enamel defects due to lower clearance than mature mice. This study provides critical evidence of age-related differences in fluoride toxicity, revealing heightened vulnerability during developmental stages. Our findings have significant public health implications, supporting the need for age-specific fluoride exposure guidelines to balance caries prevention and developmental fluoride toxicity.