Aim: To evaluate the effect of gamma radiation on bond strength and microleakage of nano-composite and nano-glassionomer, and to detect any alterations in their molecular structure due to gamma radiation. Materials and Methods: 80 specimens were used as follow; 40 specimens for shear bond strength evaluation, 20 specimens for microleakage assessment, while the remaining 20 specimens for deducing the chemical structure. For shear bond strength (SBS) test 2 mm thick wafers of dentine were sectioned and 3 mm diameter holes were drilled through the wafers. 20 specimens were restored with nano-composite and nano-glass ionomer without irradiation (Group A1, B1). The remaining 20 specimens were restored with nano-composite and nano-glass ionomer (Group A2, B2), then they were irradiated with therapeutic dose of 60 gray for 1 week (3days/week). For microleakage, 10 natural teeth with two prepared class V cavities were used. One of the cavities was restored with nano-composite while the other one with nano-glassionomer to be examined before and after gamma radiation. Spectrophotometric analysis was performed for all tested materials before and after radiation to trace any structural changes. Results: Significant increase in SBS of nano-composite after irradiation while nano-glassionomer was insignificantly increased. For microleakage no significant difference existed between the irradiated and non-irradiated groups of both materials. Conclusion: Therapeutic dose of head and neck gamma radiation had improved dentin shear bond strength of nano-composite. On the other hand, it had not an effect on shear bond strength of nano-glass ionomer and the microleakage of both tested materials. Gamma radiation did not alter the chemical structure of the tested material.