Aim: This study evaluated and compared the remineralization potential of nano-seashell, nano-pearl, and nano-hydroxyapatite (nHA) pastes versus fluoride-based toothpaste by assessing enamel surface microhardness and laser fluorescence analysis (DIAGNOdent). Materials and methods: Eighty premolars were distributed into 4 groups of 20 teeth each, according to the remineralizing agent used. Group A: nano-seashell paste; Group B: nano pearl paste; Group C: nHA paste; Group D: fluoride-based toothpaste (positive control) group. Ten teeth of each group were subjected to microhardness assessment at baseline, after demineralization for 72 hours and after 28 days of remineralization with different remineralizing materials.The rest 10 teeth of each group were assessed using the DIAGNOdent pen at baseline, after demineralization, after 14 days, and after 28 days of remineralization. The data were statistically analyzed. Results: After 28 days of remineralization, all groups showed increase in enamel surface microhardness. The highest mean percent increase of enamel microhardness was recorded in group D (458.59), followed by group B (405.97), then group A (286), with the least value recorded in group C (207.8). After 14 days remineralization, intergroup comparison of DIAGNOdent readings showed a percent decrease of laser fluorescence with no significant difference between groups. After 28 days remineralization, the greatest mean percent decrease was recorded in group A, followed by group D, then group C, with the least percent decrease recorded in group B. Conclusion: Nano-seashell, nano-pearl, nHA pastes were demonstrated to own remineralization potential comparable with fluoride-based toothpaste on initial non-cavited enamel lesions in an effective noninvasive manner.