Time-Effect Comparative Evaluation of Three Remineralizing Agents on Artificial Enamel Lesions: A SEM-EDX In Vitro Study

<b>Objective:</b> This in vitro study quantitatively compared the time-dependent remineralization potential of three professional agents on artificially induced enamel lesions using Scanning Electron Microscopy (SEM) and energy-dispersive X-ray analysis (EDX). <b>Methods:</b> Sixty extracted sound molars were randomly assigned to three groups (number = 20): G_CPP-ACP, treated with casein phosphopeptide-amorphous calcium phosphate; G_Zn-HA, treated with zinc-hydroxyapatite; and G_F-ACP, treated with fluoridated amorphous calcium phosphate. The crown of each tooth was divided into three areas: one represented the control (CTRL, sound enamel), one underwent demineralization (DEMIN, demineralized enamel), and the third one was at first demineralized and then treated with a remineralizing agent, allowing intra-sample comparison. Artificial lesions were produced by immersion in 0.1 M lactic acid (72 h). Groups were subdivided according to remineralization time (7, 14, 21, and 28 days). Samples underwent daily treatment under a pH-cycling regimen. Surface morphology and Ca/P ratios were evaluated by SEM-EDX, and data were statistically analyzed (<i>p</i> < 0.05). <b>Results:</b> All agents promoted a progressive increase in Ca/P ratio over time, confirming a time-dependent remineralization effect. At day 7, G_Zn-HA showed higher Ca/P values, but from day 14 onward, G_F-ACP produced significantly greater mineral gain than the other groups (<i>p</i> < 0.05). By day 21, G_F-ACP reached Ca/P values approaching CTRL, while G_CPP-ACP and G_Zn-HA remained at lower levels, reaching a plateau respectively at 21 and 14 days. SEM observations supported these findings: G_CPP-ACP and G_Zn-HA showed partial surface recovery, whereas G_F-ACP exhibited a compact, homogeneous enamel-like structure at 28 days. <b>Conclusions:</b> All tested agents demonstrated time-dependent remineralization, enhanced with prolonged exposure, suggesting that the time of application represents a key factor for clinical success.