A unique type of dental varnish (DV) containing surface pre-reacted glass (S-PRG) fillers of different concentrations was evaluated to determine the unpresented optimal ratio for inhibiting root dentin bio-demineralization. S-PRG DVs (10% to 40%)—10%-S, 20%-S, 30%-S, and 40%-S—were applied to bovine root dentin blocks and compared with controls—0%-f (no S-PRG) and 5%-NaF (5%-NaF). The Streptococcus mutans biofilm challenge was executed inside and outside an oral biofilm reactor for 7 days. The specimens were examined using a confocal laser scanning microscope and swept-source optical coherence tomography. Furthermore, they were observed using a scanning electron microscope and analyzed using energy-dispersive X-ray spectroscopy. The roughness (SzJIS) due to leaching of DV materials and demineralization depth were significantly less in the S-PRG groups than the control groups (p < 0.05). Complete or partially plugged dentinal tubules (DTs) were observed in 20%-S, 30%-S, and 40%-S, while wide-open DTs were observed more in controls. Cylindrical tags were present in groups containing more than 20% S-PRG. F, Na, Al, and Sr were detected in a higher percentile ratio in the 20%-S, 30%-S, and 40%-S groups compared to 0%-f and 10%-S. Nonetheless, it is suggested that incorporating 20% to 30% S-PRG fillers in DVs would be effective enough as an anti-demineralization coating, together with supplementing minerals; further evaluation is required to validate these findings.
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