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Introduction: Hypoestrogenism causes structural changes in the vaginal wall that can lead to sexual dysfunction. A reduction in vaginal wall thickness has been reported to occur after menopause, although without precise morphometry., Aim: To measure vaginal wall thickness in women with genital prolapse in normal and hypoestrogenic conditions and to correlate sexual dysfunction with vaginal wall thickness and estradiol levels., Methods: Surgical vaginal specimens from 18 normoestrogenic and 13 postmenopausal women submitted to surgery for genital prolapse grades I and II were examined. Patients were evaluated for FSH, estradiol, prolactin, glycemia, and serum TSH levels. For histological analysis, samples were stained with Masson's trichrome and hematoxylin-eosin. Sexual function was assessed by the Golombok-Rust Inventory of Sexual Satisfaction (GRISS)., Main Outcome Measures: GRISS questionnaire, histological analysis, morphometric methods, Masson's trichrome., Results: The vaginal wall was thicker in the postmenopausal than premenopausal group (2.72 +/- 0.72 mm and 2.16 +/- 0.43, P = 0.01, and 2.63 +/- 0.71 mm and 2.07 +/- 0.49 mm, P = 0.01, for the anterior and posterior walls, respectively). These thicknesses seem to be due to the muscular layer, which was also thicker in the postmenopausal group (1.54 +/- 0.44 and 1.09 +/- 0.3 mm, P = 0.02, and 1.45 +/- 0.47 and 1.07 +/- 0.44 mm, P = 0.03, for the anterior and posterior wall, respectively). The vaginal epithelium was thinner in the middle segment than in the proximal one in the posterior wall (0.17 +/- 0.07 mm, 0.15 +/- 0.05 mm, 0.24 +/- 0.09 mm, P = 0.02). There was no correlation between coital pain, vaginal wall thickness, and estradiol levels in either group., Conclusion: The vaginal wall is thicker after menopause in women with genital prolapse. In this study, vaginal thickness and estrogen levels were not related to sexual dysfunction.