This study aimed to evaluate the anticariogenic and remineralization effects of the glass ionomer dental luting cement containing nano-β-TCP in vitro. The β-Tricalcium Phosphate (β-TCP) are the components of dental enamel and bone mineral as biological apatites. In addition, β-TCP contains a significant amount of calcium and phosphate, which can promote remineralization of enamel subsurface lesions in animal and human. RelyXTM glass ionomer cement(3M/ESPE, USA) was used as dental luting cement. Film thickness, setting time, and compressive strength was measured for each group of pure glass GIC, 15% nano-β-TCP GIC. Human molars were prepared in box-shaped cavities that were filled with the GIC with and without the 15% nano-β-TCP were placed in 25ml of pH 5.0 acid buffer for 4 days at 37°C. After 4 days, longitudinal sections (1007m) were obtained through the center of each restoration. The sections were analyzed using a scanning electronic microscope (SEM) and confocal laser scanning microscopy (CLSM) to identify the change in the enamel surface. A significant difference in the CLSM images between pure GIC and nano-β-TCP-GIC. CLSM allows the demineralized surface layers of sound enamel to be visualized down to approximately 100 μm. The pure GIC specimens had a relatively thick fluorescent layer. On the other hand, the fluorescent layer of the nano-β-TCP-GIC specimens were thinner. The SEM images of micro surfaces demonstrate that nano-β-TCP-GIC is less rough than pure GIC. Therefore, the addition of nano-β-TCP enhanced protection against acid demineralization and promoted remineralization of enamel surface.