Purpose: This in vitro study was established to compare the effect of natural versus synthetic desensitizing agents on dentin permeability. Materials and Methods: Class V cavity preparation was done on the labial surface of 40 premolars for cervical dentin assessment, while for coronal dentin; class I cavity was prepared on the occlusal of 40 premolars. Cavities were etched with phosphoric acid then classified according to treatment either by Bio active glass, Pro-Argin, Moringa extract or untreated (control). The dentin permeability was done using dye penetration test with Evans blue dye. The depth of dye penetration in dentin was examined using stereomicroscope. Results: The results showed that the lowest statistically significant mean depth of dye penetration values were found with Bio active glass followed by Pro-Argin, then Moringa extract. Conclusion: Among the three treatment modalities, Bio active glass and Pro-Argin can be considered as effective materials in the management of hypersensitivity