Background Obstructing dentinal tubules is a valuable approach for managing dentin hypersensitivity. Although various agents promote dentin remineralization, direct comparisons between theobromine, bioactive glass (BAG), and nano-hydroxyapatite (Nano-HAP) under simulated oral conditions remain limited. To fill this gap, this in vitro study aimed to evaluate and compare the effectiveness of these three treatments on exposed cervical dentin. The assessment focused on their chemical, morphological, and mechanical effects on dentin. Materials and methods Forty-eight human dentin slabs were obtained from the cervical portions of twelve sound premolar teeth. Baseline Raman spectroscopy and VMH tests were done to exclude outliers. All specimens were treated with 6 % citric acid (pH 2.0) for 2 min to remove the smear layer. They randomly assigned to four groups (n = 12): artificial saliva (AS), theobromine, BAG, and Nano-HAP. Evaluations were conducted using Raman spectroscopy (phosphate peak intensity at 960cm−1), Vickers microhardness testing (VMH), and morphological assessment under scanning electron microscopy (SEM). Results Theobromine, BAG, and nano-HAP groups demonstrated a statistically significant increase in Raman phosphate peak intensity (960cm−1) and Vickers microhardness values (p < 0.05), indicating surface remineralization. In contrast, the artificial saliva group exhibited a significant decrease in phosphate peak intensity and microhardness values (p < 0.05). Conclusion All tested agents significantly enhanced the Raman phosphate peaks and microhardness values compared to the control. Nano-HAP showed the highest potential for promoting the remineralization of exposed dentin surfaces. Within the study's limitations, it can be concluded that theobromine, BAG, and nano-HAP are effective in occluding dentinal tubules.