Objective: Dental caries is a chronic infectious disease that is prevalent worldwide in all age groups. Numerous attempts have been made to develop conservative approaches to halt caries progression and restore enamel defects. This study aimed to investigate the effect of applying grape seed extract and chicken eggshell extract on the microhardness of demineralized enamel in permanent teeth. Methods: Forty-eight sound upper first premolars were used. Following demineralization with the demineralizing solution for 96 hours, they were distributed into four groups consistent with the treatment agent used: group A was treated with casein phosphopeptide amorphous calcium phosphate (as a control group), group B was treated with grape seed extract, group C was treated with chicken eggshell extract solution, and group D was treated with grape seed extract followed by chicken eggshell extract solution. Vickers microhardness measurements were performed on sound enamel, after demineralization and remineralization. Results: Paired T-test, one-way ANOVA, and Tukey’s HSD test were used for statistical analysis. Enamel microhardness was significantly reduced following demineralization (p=0.000) and significantly increased after remineralization, with group D showing the highest values (mean microhardness=218.99). Conclusion: Grape seed extract and chicken eggshell extract solutions have a synergistic effect on enamel remineralization which was interpreted from the increase in surface microhardness values.