This study aimed to assess and compare the efficacy of Nacre and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on the remineralization of enamel using surface microhardness analysis, scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) spectroscopy. Twenty human maxillary premolars extracted for orthodontic reasons were collected. Under cool water spray, the crowns were sectioned mesiodistally into buccal and palatal halves using a diamond disc. The samples were subsequently mounted in self-cure acrylic resin. The samples were then subjected to Vickers hardness testing and SEM-EDX for baseline. To simulate carious lesions, all of the samples were acid-etched with 37% phosphoric acid for 30 s in a specific area on the enamel samples and subjected to surface microhardness testing and SEM-EDX. The enamel samples were randomly assigned to Group 1: Nacre water-soluble matrix (WSM), Group 2: Nacre varnish, and Group 3: CPP-ACP for remineralization. After 21 days, remineralization assessment of the test samples was done using SMH analysis and SEM-EDX analysis. Data obtained were statistically analyzed using the one-way analysis of variance to reveal the significant differences between the groups. Tukey's test was used for post hoc comparisons. All three groups showed a significant increase in surface microhardness. All three groups showed a significant calcium and phosphorous ratio increase after remineralization. Among the three groups, the highest Ca:P ratio was seen in the Nacre WSM group (0.58) followed by the Nacre Varnish (0.57) and CPP-ACP group (0.57). SEM images of the Nacre surface revealed the presence of extensive interlocking. A layer of packed hydroxyapatite particles was formed on the surface of the nacre through surface reactions. All the groups in the present study showed some extent of remineralizing ability irrespective of the different materials and mechanisms of action. Nacre WSM showed a remarkable hardness spike close to natural enamel after demineralization.