Abstract Objective To analyze the dermal collagen of 15 postmenopausal women who had being treated with systemic estrogen replacement before and after using topical a 0.01% estrogen treatment. Methods Fifteen patients were included in this clinical trial using the systemic estrogen therapy for at least 1 year (minimum and maximum lengths of therapy were 13 and 40 months, respectively). A facial punch was performed in the preauricular area for collecting samples before and after the 16 weeks of treatment. Blood samples were also collected for estradiol level determination. The morphometric determination of epithelial and dermal thickness as well as dermal collagen were measured using a suitable software. The paired Student's t-test was used for statistical analysis. Results The epithelial and dermal thickness enhanced after the topic estrogen therapy ( P < 0.01). The amount of collagen significantly increased after 16 weeks of treatment ( P < 0.001). The estrogen levels did not significant increase after the topical therapy ( P ≥ 0.05). Conclusion Our data suggested that topical estrogen associated to systemic estrogen therapy seems to increase the expression of skin collagen amount, which may prove to be beneficial for the postmenopausal facial skin.