This study aimed to evaluate the enamel remineralization efficacy of enamel matrix derivative (EMD), experimental bioactive glass (BAG), and fluoride varnish in vitro. Artificial initial caries lesions were developed on fifty human enamel specimens using demineralization solution (pH 4.5, 37°C, 96 hours). Specimens were randomly assigned to five groups (n=10): I-5% NaF varnish (Enamelast), II-experimental 58S5 BAG+37% phosphoric acid (PA), III-EMD (Emdogain) + Ethylenediaminetetraacetic Acid (EDTA), IV-EMD+37% PA, V-Control (untreated). All remineralization agents were applied with pH cycling for seven days. The specimens were scanned by spectral domain optical coherence tomography (SD-OCT) at baseline, at demineralization, and after pH cycling. Lesion depths were measured using image analysis software (ImageJ). Lesions were evaluated using surface microhardness (SMH) and two fluorescence methods (FluoreCam and DIAGNOdent Pen [DDPen]). The data were statistically analyzed by Kruskal Wallis, Friedman, and Wilcoxon tests (α=0.05). According to SD-OCT results, fluoride varnish was found to be the most effective agent in reducing lesion depth (p=0.005). All agents increased the SMH values after pH cycling. No significant difference was found among fluoride varnish, BAG, and EMD+PA groups. These SMH values were significantly higher than EMD+EDTA and control groups (p<0.001). All groups showed lower DDPen scores compared with the control group (p<0.001), however, no significant difference was found among the remineralization agents. In FluoreCam assessment, size and intensity values of all treated groups showed improvement. However, there was no significant difference between the treatment groups in terms of FluoreCam size measurements (p=0.186). 58S5 BAG and EMD+PA have remineralization capacity as effective as fluoride varnish. EMD+PA showed better SMH and lesion intensity results than EMD+EDTA.
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