Purpose: To evaluate the clinical efficacy and safety of intravaginal application of 25 µg micronized oestradiol in postmenopausal women from the Greek population suffering from symptoms related to vaginal atrophy. Materials and Methods: 91 women suffering from vaginal dryness, vaginal itching and dyspareunea were treated with 25 µg 17β-oestradiol vaginal tablets. The duration of treatment was 12 weeks. During the first two weeks the women inserted one vaginal tablet intravaginally once daily. Thereafter, the women inserted one tablet twice per week with at least a 3-day interval between treatments to maintain therapeutic response for ten weeks. Efficacy was evaluated by the relief of vaginal symptoms and safety by the con­centrations of serum oestradiol (E2) and follicular-stimulating hormone (FSH). Pretreatment and post-treatment findings were com­pared and each patient served as her own control. Results: The rates of symptoms of vaginal dryness, vaginal itching and dyspareunea showed statistically significant differences over the course of the trial (Cochran Q test, p < 0.001). No one complained of vaginal dryness and vaginal itching after four and six weeks of treatment respectively, while in one patient the sensation of dyspareunea remained constant after the fourth week of treatment. Despite the statistically significant increase in blood oestradiol levels in relation to baseline values (ANOVA model of repeated measures, p < 0.001), these levels were within the normal range for postmenopausal women. Also, serum FSH levels were statistically significantly reduced from 47.4 mIU/mL at entry into the study to 45.5 mIU/mL after two weeks of treatment (dependent samples t-test, p < 0.003), but were clearly within the postmenopausal range. Conclusions: The twice-weekly local single treatment with vaginal tablets containing 25 µg of 17β-oestradiol was effective and safe for the relief of symptoms related to atrophic vaginitis in postmenopausal women from the Greek population.