Sexual steroids in urogynecology.

The decline in sex hormone levels that accompanies the menopause has substantial effects on the tissues of the urogenital system, leading to atrophic changes. These changes can have negative effects on sexual and urinary function. The authors evaluate the repercussion of hypoestrogenism and sexual steroids on some elements of the pelvic floor and lower urinary tract. They summarize their research work and review significant published papers. They emphasize the changes in urinary mucosae, periurethral vessels, muscular layer, connective tissue, gene expression, autonomic nervous system receptors, as well as the main clinical aspects involved.

Estrogen effects on the vaginal pH, flora and cytology in late postmenopause after a long period without hormone therapy.

In this report we evaluated the action of conjugated equine estrogens (CEE) on vaginal symptoms, cytology, pH, and flora in late postmenopausal women without any previous hormone therapy. The study was a randomized, double-blind, placebo-controlled trial with 48 late postmenopausal women who received placebo or unopposed CEE (0.625mg/day of CEE orally) during three months of treatment. Vaginal and sexual complaints were evaluated through daily diary cards. We analyzed vaginal changes through cytology and pH measurements. After three months of treatment, 20% of placebo-treated patients and 80% of the CEE-treated patients reported improvement in vaginal dryness and irritation. In the latter group, the vaginal cells and Lactobacillus increased and the vaginal pH decreased, without other changes in sexual complaints. We concluded that estrogen ameliorated the genital tract of late postmenopausal women without any previous hormone therapy.

Effects of tamoxifen on the breast in the luteal phase of the menstrual cycle.

OBJECTIVES: The effect of tamoxifen on cyclic mastalgia and on chemoprophylaxis against breast cancer is little known, mainly due to the difficulties in studying the normal human gland. We proposed to evaluate the mitotic index and the nuclear volume of the lobule of women medicated with tamoxifen only during the luteal phase of the menstrual cycle in order to observe the effect of tamoxifen on the normal human mammary gland. METHODS: Twenty-four premenopausal women with fibroadenoma diagnosed via biopsy were studied. The phase of the cycle was determined by the date of menstruation and serum progesterone level in the luteal phase (> or = 3 ng/ml). The patients admitted to the study and were given written informed consent to participate in the investigation, which was previously approved of by the hospital Ethics Committee. Patients were divided at random into two groups: Group I consisted of 12 untreated women (control) and Group II consisted of 12 patients treated with 20 mg/day tamoxifen for 10 consecutive days beginning on the 13th day of the menstrual cycle. In both groups, the patients were submitted to biopsies of the nodule and of a 1-cm3 fragment of adjacent mammary parenchyma between the 23rd and 26th day of the cycle. The mitotic index (number of mitoses/1000 nuclei counted) and mean nuclear volume (mean of 10 nuclear volumes for each case) were measured. RESULTS: No mitosis was observed in Group II. There was a reduction in the mean nuclear volume in Group II (Mann-Whitney test). CONCLUSIONS: Tamoxifen, when administered only during the luteal phase of the menstrual cycle, significantly reduces the nuclear volume and mitotic activity of the epithelium. This data demonstrates an antagonistic action of tamoxifen on estrogen even when administered for short periods of time.

Effect of a half dose of tamoxifen on proliferative activity in normal breast tissue.

OBJECTIVES: To investigate the proliferative activity of the mammary gland epithelium and plasma levels of progesterone, estradiol, prolactin, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and sex hormone-binding globulin (SHBG) in premenopausal women treated with 10 and 20 mg of tamoxifen (TAM) for 22 days. PATIENTS AND METHODS: A randomized double-blind study was performed with 43 premenopausal women with a diagnosis of fibroadenoma of the breast. The patients were divided into three groups: A (n = 15, placebo); B (n = 15, TAM 10 mg/day) and C (n = 13, TAM 20 mg/day). They started taking an oral dose of TAM or placebo on the very first day of the menstrual cycle. Lumpectomy was performed on the 22nd day of therapy. Normal breast tissue samples were collected during surgery, immediately immersed in 10% buffered formalin, processed for routine histology and immunohistochemistry for proliferating cell nuclear antigen (PCNA) detection. Two peripheral blood samples were collected, both on the 22nd day of the menstrual cycle, in order to evaluate the hormone levels. PCNA expressing epithelial cells were quantified by using a digital program Kontron Image System KS-300 in 1000 cells (400 x ). RESULTS: The percentage of cells expressing PCNA was significantly higher in the group receiving placebo (group A, 50.3%) when compared to groups receiving TAM 10 or 20 mg/day (group B, 24.1%; and group C, 23.2%, respectively) (P < 0.001). Differences between groups B and C were not significant. Levels of progesterone, estradiol and SHBG were significantly higher in B and C groups compared to group A. Increasing concentrations of FSH (P < 0.0045) and lower levels of prolactin (P < 0.0055) were only found in the group receiving 20 mg/day of TAM (group C). CONCLUSIONS: A 22-day TAM therapy, either with 10 or 20 mg/day, significantly reduced the PCNA expression and therefore the proliferative activity of the normal human breast tissue. Increasing levels of estradiol, progesterone and SHBG were associated with TAM therapy at 10 or 20 mg/day. However, a significant change of the level of FSH and prolactin was reached only with a 20-mg/day dose.

Effects of the association of androgen/estrogen on the bladder and urethra of castrated rats.

PURPOSE: To study the effects of methyltestosterone, isolated or associated to estrogens, on the bladder and urethra of castrated adult rats. MATERIAL & METHODS: A total of 59 castrated animals were studied. They were divided into the following groups: I--placebo; II--equine conjugated estrogens; III--methyltestosterone and conjugated estrogens; IV--methyltestosterone. After 28 days of medication the animals were sacrificed and bladder and urethra cuts were obtained for the evaluation of the number of vessels, the thickness of the epithelia, and the quantity of collagen and muscular fibers. RESULTS: The group receiving the androgen/estrogen association presented a higher number of vessels, epithelial thickness and quantity of muscular fibers (p < 0.05). A smaller quantity of collagen fibers was observed in the group utilizing isolated conjugated estrogen (p < 0.05). CONCLUSION: We concluded that the association of androgen/estrogen positively modifies important parameters in the urinary continence mechanism. Therefore, it could constitute an option for hormone replacement in postmenopausal stress urinary incontinence cases.

Histomorphometric aspects of adult castrated rat endometrium after the use of estrogen, progesterone and tamoxifen.

The aim of this study was to analyse the morphologic and morphometric aspects of endometrium in rats receiving hormone replacement therapy with conjugated equine estrogen (CEE), medroxyprogesterone acetate (MPA) and tamoxifen (TMX). Thirty-five adults rats, 2-3 months of age were ovariectomized four days prior to using the drugs. Rats were divided according to the following treatments for 60 days: CEE (50 microg); CEE/MPA (50 microg/2 mg); MPA (2mg); TMX (250 microg); vehicle (propyleneglycol). Fragments of endometrium were removed and analysed by light microscopy. The endometrium suffered evident morphologic modifications under the action of hormones and TMX. The endometrium was significantly thicker in the CEE, CEE/MPA and TMX group when compared to the control, however the MPA group showed no differences when compared to the control group.

Ultrastructural aspects of the postmenopausal endometrium after oral or transdermal estrogen administration.

In this report we examined the ultrastructural features of the postmenopausal endometrial cells of women treated with different doses of conjugated equine estrogen (CEE), or transdermal 17beta-estradiol. Eight women with uterine prolapse and at least 5 years of menopause were randomly divided into four groups and treated as follows: (I) no hormonal treatment; (II) 0.625mg/day of CEE orally; (III) 1.25mg/day of CEE orally; (IV) 50microg/day of 17beta-estradiol transdermally. Hormones were administered for 28 days followed by vaginal hysterectomy. Fragments of the endometrium were prepared for transmission electron microscopic analysis. We observed that the postmenopausal endometrium of the untreated group was atrophic with lined superficial epithelial cuboidal cells. The presence of gland and stroma cells with clear cytoplasm containing few organelles and heterochromatin nuclei were also observed. On the contrary, the endometrium of the group that received 0.625mg/day of CEE showed signs of proliferative cells such as the presence of numerous organelles in the cytoplasm and euchromatic nuclei. All of the proliferative effects on the endometrium were more pronounced in the groups that received 1.25mg/day of CEE and 50microg/day of transdermal 17beta-estradiol. We concluded that the ultrastructural proliferative changes of the postmenopausal endometrium induced by 1.25mg/day of CEE were similar to 50microg/day of transdermal 17beta-estradiol.

Clinical and urodynamic evaluation of women with detrusor instability before and after functional pelvic floor electrostimulation.

Detrusor instability is the second most frequent cause of female urinary incontinence. There are many therapeutic options, including non-invasive and surgical procedures. In this study, we evaluated the effects of pelvic floor vaginal electrostimulation using equipment designed in our institution, over three consecutive months, for treatment of 29 women with detrusor instability. After treatment 22 patients (76%) considered themselves cured or symptomatically improved; seven patients (24%) had no change in symptoms after therapy. There was objective cure and improvement in ten (34.5%) and in eight (27.5%) patients, respectively, and the urodynamic parameters did not change in 11 patients (38%). Electrical stimulation resulted in a gradual decrease in the number of urinary leakage episodes and increase in maximum cystometric capacity in first desire to void and in urinary volume.

A morphologic and morphometric study of the vesical mucosa and urethra of castrated female rats following estrogen and/or progestogen replacement.

PURPOSE: The aim of this study was to analyze the morphologic and morphometric changes in the urethra and vesical mucosa following hormonal replacement METHODS: We analyzed the changes in the urethra and vesical mucosa of 35 castrated adult female rats that had been subjected to estrogen and/or progestogen replacement. RESULTS: Estrogen replacement, whether or not accompanied with progestogen replacement, provoked metaplasia, hyperplasia and an increased occurrence of stratified epithelia. In the proximal urethra hypoestrogenism caused a higher occurrence of pseudo-stratified and transition epithelia, whereas in the urethral-vesical junction it caused a higher frequency of pseudo-stratified epithelia. The thickness of the epithelium increased following estrogen replacement whereas only a trend towards an increase of the propria lamina thickness was identified. Nuclear volume was only altered in the bladder epithelium. CONCLUSION: Estrogen replacement acted both morphometrically and morphologically on the lower urinary tract.

Morphologic and morphometric study of the breast parenchyma of rats in persistent estrus treated with tamoxifen and conjugated estrogens.

PURPOSE: To evaluate the morphological and morphometric alterations produced by tamoxifen and conjugated estrogens in the mammary epithelium of rats in persistent estrus. METHODS: 33 adult female rats with persistent estrus induced by 1.25 mg testosterone propionate were divided at random into three groups: group I (n=12), receiving only water and used as a control; group II (n=10), treated with 500 microg tamoxifen daily; group III (n= 11), treated with 30 microg conjugated estrogens daily. The first abdominal-inguinal pair of breasts was extirpated and processed for morphological and morphometric study. Data were analyzed statistically by the Kruskal-Wallis rank analysis of variance (p<0.05). RESULTS: The morphological study revealed signs of epithelial atrophy and the morphometric study showed a significant reduction in mean number of ducts and alveoli in groups II (10.1 and 1.9, respectively) and III (11.1 and 3.5, respectively) compared to the control group 1 (25.0 and 6.6, respectively). There was no significant difference between groups II and III. CONCLUSIONS: The present results indicate that, at the doses and during the time of treatment used, both tamoxifen and conjugated estrogens induced atrophy of the mammary epithelium of rats in persistent estrus.

Quantitative evaluation of collagen and muscle fibers in the lower urinary tract of castrated and under-hormone replacement female rats.

OBJECTIVE: To evaluate the number of collagen and muscle fibers in the muscle layer of the urethra and in the bladder wall of castrated and under-hormone replacement female rats. METHOD: We studied 37 castrated female rats assigned to the following groups: Group C (n=9): received no medication; Group P (n=8) was given 0.1 ml of placebo, subcutaneous (SC) route; Group E (n=10): 17beta-estradiol, 10 microg/kg/day, SC route; Group PR (n=9): medroxyprogesterone acetate. 0.2 mg/kg/day, SC route; Group E+PR (n=9): association of 17beta-estradiol and medroxyprogesterone acetate. Sections were taken from the bladder wall and from the middle third of the urethra, and the specimens were stained with picrosirius for collagen and muscle fiber identification. RESULTS: Groups C and P showed a similar amount of collagen in the bladder and in the urethra, however greater than the other groups. Group E showed the smallest number of collagen fibers in the urethra. Groups E and E+PR presented a larger number of muscle fibers in the bladder. Group PR presented a larger number of muscle fibers than groups C and P, however smaller than groups E and E+PR. In the muscle layer of the urethra, the number of collagen fibers was smaller in Group E than in all the other groups, which were similar among one another. In regard to the urethral muscles, Group E was found to present the largest number of muscle fibers as compared to the other groups analyzed, while Group PR showed a significant decrease in the muscle layer, even in relation to the groups that were given no hormone medication. CONCLUSION: Estrogens significantly decrease the amount of collagen fibers, increase the amount of muscle fibers and determine a significantly decreased collagen/muscle ratio in both the detrusor muscle and in the urethral muscle layer. It is also noticed that isolated progestogen decreases the amount of collagen fibers and increases the number of muscle fibers in the detrusor muscle, but with less intensity than replacement with estrogens alone. It neither alters the number of collagen fibers nor decreases the muscle fibers in the muscle layer of the urethra, with increased collagen/muscle ratio in that structure. Finally, the estrogen-progestogen combination determines significantly decreased collagen fibers and increased muscle fibers in the detrusor muscle, causing no alteration to the collagen or muscle fibers in the muscle layer of the urethra.

Urodynamic and clinical evaluation of postmenopausal women with stress urinary incontinence before and after cyclic estrogen therapy.

OBJECTIVE: The purpose [corrected] of this study was to evaluate the effects of isolated cyclic estrogen therapy in menopausal women with stress urinary incontinence, and thus without the effects of progesterone. METHODS: Nineteen menopausal patients with stress urinary incontinence were selected and submitted to anamnesis and physical, gynecological and urodynamic examinations. The group was homogeneous in relation to parity, body mass index and degree of urogenital prolapse. All the patients received conjugated equine estrogens orally, at a dose of 0.625 mg, for 21 days each month. After three months the clinical and urodynamic evaluations in relation to urine loss, were performed again. RESULTS: Of the patients 57.9% were satisfied with the treatment. The urodynamic parameters remained unaltered in 36.85% of the patients. CONCLUSION: Our results show that estrogen is important for stress urinary incontinence in postmenopause, specially in patients without cystocele or with cystocele of degree I or II.

Does electrical stimulation of the pelvic floor make any change in urodynamic parameters? When to expect a cure and improvement in women with stress urinary incontinence?

OBJECTIVE: Our study aimed at determining the effects of pelvic floor electrical stimulation assessed by the number of leakages per day recorded in a voiding diary over 90 days of treatment and urodynamic parameters. STUDY DESIGN: This prospective study was carried out with 34 patients presenting stress urinary incontinence who were treated and evaluated by voiding diaries and urodynamic tests. The primary outcome measure was the number of leakages during the 90 days of treatment. Urodynamic tests were performed before and after treatment. RESULTS: In our series, average and maximum flow rates and residual urine volume were within normal range in all subjects before and after treatment. Maximum urethral closure pressure and functional profile length on urethral pressure profiles did not change after treatment. In the cystometry, bladder capacities at the first (p < 0.0082) and maximum sensations (p < 0.01) improved significantly after treatment. During the 90 days of treatment, we observed a gradual drop in the number of leakages. This decrease began around day 22. It dropped in half around day 45, tending to zero close to day 90 of treatment (p < 0.01). CONCLUSIONS: The number of incontinent leakage dropped to half around the 8th week, and on average, there was a tendency of the patients to be cured after the 12th week of treatment. At urodynamic studies we observed a significant increase in bladder capacity at the first desire to void and in the maximum cystometric capacity.

Mechanical properties of polypropylene mesh used in pelvic floor repair.

The aim of this study was the comparison of the stiffness of different meshes under two types of mechanical tests. Five different mesh types were mechanically tested. The methods used consisted on uniaxial tension test (tensile stiffness) and tape ring tests, experimental continuous compression of the mesh loops (flexural stiffness). The most significant difference of tensile stiffness behaviour appears between Aris and TVTO. From the analysis of the experimental data, we divided the flexural stiffness, in two main groups. The first group includes Auto Suture and Aris meshes. The two meshes seem to have a similar flexural behaviour. The second group includes TVTO, Uretex and Avaulta. The difference between these two groups is clearly evident comparing TVTO and Aris. This study shows that there are significant differences on the mechanical properties between urogynecology meshes.

Impact of pregnancy and childbirth on female rats' urethral nerve fibers.

This study aims to evaluate the urethral nerve fibers of adult female rats during pregnancy and after vaginal birth, cesarean section or simulated birth trauma. For immunohistochemical analysis of nerve fibers, 70 female rats were distributed in seven groups of ten female rats: group 1, control; group 2, pregnant; group 3, cesarean section; group 4, vaginal birth; group 5, virgin female rats with simulated birth trauma; group 6, cesarean section followed by simulation of birth trauma; and group 7, vaginal birth followed by simulation of birth trauma. The number of nerve fibers in groups 1, 2, and 3 were significantly higher than the other groups. Pregnancy and cesarean section did not cause alterations in the nerve fibers number. Vaginal birth and simulated birth trauma significantly decreased the number of nerve fibers in the female rats' middle urethra.

The impact of pregnancy and childbirth in the urethra of female rats.

The aim of this study was to evaluate the modifications in the amount of collagen, muscular, and elastic fibers in the mid-urethra of adult female rats during the pregnancy and after the natural childbirth, cesarean, and after simulated trauma of childbirth. The authors evaluated the histomorphometric aspects (collagen, muscular, and elastic fibers) in the mid-urethra of 70 animals distributed in seven groups: group 1 (n = 10)--control, group 2 (n = 10)--pregnant female rats, group 3 (n = 10)--female rats submitted to cesarean, group 4 (n = 10)--female rats with natural childbirth, group 5 (n = 10)--virgin female rats with simulated trauma of childbirth, group 6 (n = 10)--female rats submitted to cesarean followed by simulation of childbirth trauma, and group 7 (n = 10)--female rats with natural childbirth followed by simulation of childbirth trauma. The average concentration of collagen and elastic fibers and the collagen/muscular fiber correlation in groups 1, 2, and 3 were similar and significantly inferior to groups 4, 5, 6, and 7. The average of muscular fibers was similar in groups 1, 2, and 3 and significantly superior to groups 4, 5, 6, and 7. Pregnancy and cesarean did not induce alterations in collagen, muscular, and elastic fibers. However, the vaginal delivery and simulation of childbirth trauma determined the decrease in muscular fibers and the increase in collagen and elastic fibers and the correlation collagen/muscular fiber.

Urodynamic and ultrasonographic evaluation after continence surgery.

Our objective was to evaluate urodynamic and ultrasonographic findings after continence surgery. The study consisted of three groups of women according to the surgery performed: group I (Burch colposuspension) with 12 patients; group II (Kelly-Kennedy plication) with 10 patients; and group III (Gittes surgery) with 9 patients. Urodynamic study was done preoperatively and after surgery (on the 7th and 30th postoperative days, and at least 6 months after surgery) and ultrasonography of the bladder neck was performed to evaluate its position in relation to the inferior edge of the pubic symphysis and its mobility, both preoperatively and after surgery (30th postoperative day). All patients remained continent. We observed an increase in the first desire to void and maximum cystometric capacity after 6 months in groups I and II, respectively. There was no change in the urethral closure pressure profile in the three groups. Elevation of the bladder neck and decrease of its mobility were found by ultrasonography. Urinary continence after surgery is not the result of alterations in the urethral closure pressure profile, but rather of an elevation in the bladder neck and limitation of its mobility, which probably improves the abdominal pressure transmission rate to the proximal urethra.

Effect of estrogen-progestogen hormonal replacement therapy on periurethral and bladder vessels.

This study assessed the effect of hormone replacement therapy using estrogens and/or progestogens on the number of vessels in the proximal and distal urethra, vesicourethral junction and bladder of castrated adult female rats. Forty-five virgin adult rats (Rattus norvegicus albinus) castrated for at least 30 days were used. They were assigned to five groups; group I (control) received no medication; the others received via the subcutaneous route, respectively, 17-beta-estradiol (group II), medroxyprogesterone acetate (group III), a maize oil and benzyl acid solution - placebo (group IV) and 17-beta-estradiol combined with medroxyprogesterone acetate (group V), for a minimum of 28 days. Increased vascularization throughout the urinary tract, except in the distal urethra, was found following estrogen replacement alone. In the group that received combined estrogens and progestogens, no increase was found. It was concluded that estrogen replacement in castrated rats significantly increased the number of vessels in the lower urinary tract.

Changes in the lower urinary tract in continent women and in women with stress urinary incontinence, according to menopausal status.

The aim of this study was to assess the impact of the postmenopausal period on clinical and urodynamic parameters and on the mobility of the bladder neck in continent women and in women with stress urinary incontinence. Fifty-seven postmenopausal women were studied: 30 were continent and 27 had stress urinary incontinence. They were subdivided according to postmenopausal stage into groups A (<5 years) and B (>5 years). Five years was a good marker to separate those women with mild and severe estrogen deficiency. Fifteen premenopausal incontinent women were selected for bladder neck ultrasound as controls. All underwent history, general physical and gynecologic examinations, LH and FSH determinations, type 1 urine and uroculture, circadian voiding diary, cotton-swab test, bladder neck ultrasound and urodynamic investigations. Analysis of the voiding diaries revealed a higher frequency of daytime micturition in both groups of incontinent patients than in the continent ones. Increased bladder neck mobility was also found, both in the cotton-swab test and an ultrasound, in group A and an ultrasound in the premenopausal incontinent women. Urodynamic investigation showed decreased bladder capacity at the first micturition urge, as well as decreased urinary volume in the group A patients compared to the continent ones. Decreased urethral closure maximum pressure was also found in group B patients in relation to the continent ones. We concluded that the effect of hypoestrogenism, manifested postmenopause, causes changes in the lower urinary tract of women, particularly those who are incontinent.

Analysis of collagen in parametrium and vaginal apex of women with and without uterine prolapse.

Our objective was to compare the amount of collagen in parametrium and vaginal apex between women with uterine prolapse at pre- and postmenopause, and in women without prolapse. The study included 22 premenopausal women without prolapse (group A), 10 premenopausal women with prolapse (group B), and 23 postmenopausal women with prolapse (group C) (total 55). Patients in group A underwent abdominal hysterectomy for uterine leiomyoma, and patients in groups B and C underwent vaginal hysterectomy. During the surgical procedure we obtained biopsies from the lateral parametrium and vaginal apex. The tissue was stained for histological analysis with picrosirius. We observed a lower amount of collagen in the parametrium of women with uterine prolapse, both in menacme and in postmenopause, than in the parametrium of women without prolapse. We observed no statistically significant difference in vaginal apex between the groups.