This study aimed to evaluate the effect of a novel bioactive glass (BAG)-containing dentin adhesive on the permeability of demineralized dentin. Bioactive glass (85% SiO2, 15% CaO) was fabricated using the sol-gel process, and two experimental dentin adhesives were prepared with 3 wt% silica (silica-containing dentin adhesive; SCA) or BAG (BAG-containing dentin adhesive; BCA). Micro-tensile bond strength (μTBS) test, fracture mode analysis, field-emission scanning electron microscopy (FE-SEM) analysis of adhesive and demineralized dentin, real-time dentinal fluid flow (DFF) rate measurement, and Raman confocal microscopy were performed to compare SCA and BCA. There was no difference in μTBS between the SCA and BCA (p > 0.05). Multiple precipitates were evident on the surface of the BCA, and partial occlusion of dentinal tubules was observed in FE-SEM of BCA-approximated dentin. The DFF rate was reduced by 50.10% after BCA approximation and increased by 6.54% after SCA approximation. Raman confocal spectroscopy revealed an increased intensity of the hydroxyapatite (HA) peak on the dentin surface after BCA application. The novel BAG-containing dentin adhesive showed the potential of both reducing dentin permeability and dentin remineralization.