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Objective: To explore the effect of topical estrogen therapy on the expression of estrogen receptor β (ERβ) in bladder tissue of female patients with overactive bladder (OAB)., Methods: A total of 58 female OAB patients who were treated in the Affiliated Hospital of Qinghai University were included in this retrospective study. The patients were divided into an estrogen group (28 cases) and a tolterodine group (30 cases). In the estrogen group, patients received topical vaginal estrogen treatment (0.5 mg daily) for 3 consecutive weeks and this was followed by a one-week interval. In the tolterodine group, patients received tolterodine (4 mg once daily) for 3 consecutive weeks and this was followed by a one-week interval. All patients underwent cystoscopy and completed Overactive Bladder questionnaire short form (OAB-q SF). The expression of ERβ in the bladder tissue was detected by immunohistochemistry., Results: After 12 weeks, there was no statistical difference in the OAB-q scores between the tolterodine and estrogen groups. However, tolterodine treatment significantly improved urinary incontinence than estrogen treatment ( P = 0.03). After 12 weeks of estradiol treatment, the expression of ERβ in the bladder tissue was significantly higher than that in the tolterodine group ( P < 0.05)., Conclusion: Topical estrogen therapy ameliorates OAB in female patients, and this may be related to improved ERβ expression in the bladder mucosa., Competing Interests: None., (AJTR Copyright © 2023.)