To study the effect of incorporating chitosan and fluoride-loaded chitosan nanoparticles into a glass-ionomer cement (GIC) to prevent secondary caries. A standard cervical cavity (mesio-distal width 6mm, cervico-occlusal width 2mm, and depth 2mm) was prepared on 30 molars for the following restoration groups: group 1, conventional GIC restoration; group 2, chitosan (10%) modified GIC restoration; group 3, fluoride loaded chitosan nanoparticles (10%) modified GIC restoration. The restored teeth were subjected to 1,500 thermal cycles before undergoing a multi-species cariogenic biofilm challenge. The restored teeth were examined by micro-computed tomography (micro-CT), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM/EDX). Data were analyzed by the one-way ANOVA, Tukey HDS, Kruskal Wallis, and Dunn's test. Micro-CT determined outer lesion depths for groups 1-3 were: 614 ± 20μm, 589 ± 17μm, and 560 ± 19μm respectively. Both modifications with chitosan and fluoride-loaded chitosan nanoparticles significantly affected outer lesion depth (p < 0.05). The modification with fluoride-loaded chitosan nanoparticles statistically significantly decreased the outer lesion depth compared to all other groups (p < 0.05). SEM/EDX showed an increase of calcium, phosphorus, and fluoride at the root dentine adjacent to the restoration in groups 2 and 3 (modified GIC). This increase was statistically significantly higher in the group modified with fluorine-loaded nano chitosan particles compared to the other groups (p < 0.05). Incorporation of 10% chitosan and 10% fluoride-loaded chitosan nanoparticles into GIC restorative material can prevent secondary root caries development. 10% fluoride-loaded chitosan nanoparticles were more effective. Glass ionomer cement modified with fluoride-loaded chitosan nanoparticles may be a promising restorative material in pediatric and preventive dentistry due to their controlled release properties.
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