Background: This study assessed the efficacy of novel herbal-based toothpaste in reducing dentin hypersensitivity compared to commercially available toothpaste. Materials and Methods: Thirty dentin discs of 1.0± 0.1mm thickness were prepared and assigned into two main groups: Group 1 (n=15) samples without acid challenge. Group 2 (n=15) samples with acid challenge. Each group was subdivided into three subgroups according to the toothpaste: Subgroup A: discs treated with 0.5% Ashwagandha (n=5). Subgroup G: discs treated with 0.5% Ginger (n=5). Subgroup F: discs treated with Fluoride-based toothpaste (+ve control) (n=5). Samples were evaluated for the percentage of opened dentinal tubules per surface area using a Scanning Electron Microscope (SEM) and Image J Analysis at baseline, after etching with 40% citric acid for 30 seconds, and after 14 days of the treatment regimen. Results: The highest percentage of opened dentinal tubules per surface area was recorded in the subgroup G1 (7.23±2.45), followed by F1 (6.47±3.15), and then A1 (5.9±1.7) with no statistically significant difference (p = 0.685). However, the highest percentage of opened dentinal tubules per surface area was recorded in the subgroup A2 (2.59±0.96), followed by G2 (2.34±0.81), then F2 (0.39±0.18) with a statistically significant difference (p=0.003). Group 2 showed statistically significantly higher values regarding the percentage of opened dentinal tubules per surface area than Group 1. Conclusion: All the toothpastes tested successfully treated dentin hypersensitivity concerning decreased opened dentinal tubules per surface area. Ginger and Ashwagandha were efficient as natural products that could successfully replace commercial products in managing dentin hypersensitivity