The objective of this study was to evaluate the effects of irradiation on microhardness of dentin. Dentin blocks from the cervical region of bovine incisors were treated as follows: (1) no irradiation; (2) irradiation of specimens up to 60 Gy (2 Gy/day, 5 days/week); (3) no irradiation, but fluoridation of specimens for 5 min/d; (4) irradiation of specimens and daily fluoridation. Knoop hardness number (KHN) of the control specimens was 62.63 ±14.75 (mean ± SD). This was significantly different from the irradiated dentin samples (8.74 ± 2.59 KHN). Hardness of the fluoridated dentin specimens was 11.19 ± 1.95 KHN in the non‐irradiated group and 10.03±2.76 KHN in the irradiated groups, respectively. Within the limitations of an in vitro study, it is concluded that dentin is severely affected by irradiation. This could be an explanation for the frequently observed side‐effects of irradiation like loss of enamel, gap formation at the amelodentinal junction, and caries of the cervical region. Fluoridation with acidic gels decreases microhardness of dentin surface, and does not prevent softening due to radiation, when saliva is absent.