Endometriotic Patients Research Articles

Ceratonia siliqua L. pod Effects on Viability Gene Expression of Endometrial Mesenchymal Stromal/Stem Cells Isolated from Women with Endometriosis-Associated Infertility.

This study aims to investigate the effects of carob (Ceratonia siliqua L.) pod extract (CPE) on the viability of human endometrial mesenchymal stromal/stem cells (EnMSCs) and its impact on mRNA and protein expressions of DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B), histone deacetylase 1 (HDAC1), matrix metalloproteinase-2 (MMP2), and cyclooxygenase-2 (COX-2) in endometriotic patients. In this experimental study, EnMSCs were derived from endometrium of patients with ovarian endometrioma (OMA-EnMSCs group) and deep infiltrative endometriosis (DIE) samples of 10 endometriosisassociated infertility (EAI) women (E-EnMSCs group) and compared to EnMSCs derived from the endometrium of an endometriosis-free, normal woman as the control group (C-EnMSCs). The metabolic activity of the control and case groups were evaluated by treating them with different concentrations of CPE. Cell viability was analysed by MTT. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to evaluate the expression of specific genes at the mRNA and protein levels, respectively. Treatment with 0.8 and 2 μg/mL of CPE downregulated COX-2 and HDAC1 in the E-EnMSC group compared to the C-EnMSCs group. Treatment with 0.8 μg/mL of CPE also decreased MMP2 and DNMT3B gene expressions. The COX-2 and DNMT3A genes were significantly upregulated after treatment with 2 μg/mL of CPE. Expressions of the COX-2, HDAC1, DNMT1, DNMT3A, and DNMT3B peptides decreased in the all three groups after treatment with 0.8 and 2 μg/mL of CPE. Gas chromatography-mass spectroscopy (GC-MS) analysis of CPE identified 14 bioactive compounds. Molecular docking showed the best position of each bioactive compound on the different target proteins that are involved in the process of apoptosis in EnMSCs. In vitro and in silico analyses of CPE bioactive compounds show that they may downregulate the cell inflammatory pathway involved in the pathophysiology of endometriosis.

Questionnaire-based screening of adolescents and young adult women can identify markers associated with endometriosis.

Do adolescents and young adult women (YAW) with histologically proven endometriosis present a specific clinical history? Questionnaire screening of adolescents and YAW can identify clinical markers associated with histologically proven endometriosis. Some validated questionaries can contribute to an earlier endometriosis diagnosis in adults. None of these scores, however, have been validated for adolescents or YAW. This was an observational cross-sectional study using prospectively recorded data performed between January 2005 and January 2020 in a single university tertiary referral centre for endometriosis diagnosis and management. After a thorough surgical examination of the abdomino-pelvic cavity, women with histologically proven endometriosis were allocated to the endometriosis group, and symptomatic women without evidence of endometriosis were allocated to the endometriosis-free control group. The endometriotic patients were allocated into two sub-groups according to their age: adolescent (≤20 years) and YAW (21-24 years). Adolescents and YAW ≤24 years of age were operated for a symptomatic benign gynaecological condition with signed informed consent. A standardized questionnaire was prospectively completed in the month before the surgery and included epidemiological data, pelvic pain scores, family history of endometriosis, and symptoms experienced during adolescence. The study searched for correlations by univariate analysis to determine clinical markers of endometriosis in adolescents and YAW compared with endometriosis-free control patients. Of the 262 study participants, 77 women were adolescents (≤20 years of age) and 185 patients (70.6%) were YAW. The endometriosis group included 118 patients (45.0%) and 144 (55.0%) were assigned to the control group. A family history of endometriosis, absenteeism from school during menstruation, history of fainting spells during menstruation, and prescription of oral contraceptive pills for intense dysmenorrhea were significantly more frequently observed in the endometriotic patients. The prevalence and mean pain scores for dysmenorrhea, deep dyspareunia, non-cyclic chronic pelvic pain and gastrointestinal and lower urinary tract symptoms were significantly greater in the endometriosis group, as was experienced rectal bleeding. The study was performed in a single referral centre that treats patients with potentially more severe disease. This questionnaire was evaluated on a population of patients with an indication for endometriosis surgery, which can also select patients with more severe disease. Women with asymptomatic endometriosis were not considered in this study. These factors can affect the external validity of this study. Patient interviews are relevant to the diagnosis of endometriosis in adolescents and YAW. Combined with imaging and clinical examination, this approach will enable earlier diagnosis and treatment, while remaining non-invasive and rapid. The study received no funding from external sources. There are no conflicts of interest. N/A.

Horizons in Endometriosis: Proceedings of the Montreux Reproductive Summit, 14-15 July 2023.

Endometriosis is a complex and chronic gynaecological disorder that affects millions of women worldwide, leading to significant morbidity and impacting reproductive health. This condition affects up to 10% of women of reproductive age and is characterised by the presence of endometrial-like tissue outside the uterus, potentially leading to symptoms such as chronic pelvic pain, dysmenorrhoea, dyspareunia, and infertility. The Montreux summit brought a number of experts in this field together to provide a platform for discussion and exchange of ideas. These proceedings summarise the six main topics that were discussed at this summit to shed light on future directions of endometriosis classification, diagnosis, and therapeutical management. The first question addressed the possibility of preventing endometriosis in the future by identifying risk factors, genetic predispositions, and further understanding of the pathophysiology of the condition to develop targeted interventions. The clinical presentation of endometriosis is varied, and the correlation between symptoms severity and disease extent is unclear. While there is currently no universally accepted optimal classification system for endometriosis, several attempts striving towards its optimisation - each with its own advantages and limitations - were discussed. The ideal classification should be able to reconcile disease status based on the various diagnostic tools, and prognosis to guide proper patient tailored management. Regarding diagnosis, we focused on future tools and critically discussed emerging approaches aimed at reducing diagnostic delay. Preserving fertility in endometriosis patients was another debatable aspect of management that was reviewed. Moreover, besides current treatment modalities, potential novel medical therapies that can target underlying mechanisms, provide effective symptom relief, and minimise side effects in endometriotic patients were considered, including hormonal therapies, immunomodulation, and regenerative medicine. Finally, the question of hormonal substitution therapy after radical treatment for endometriosis was debated, weighing the benefits of hormone replacement.

Assessment of Differential Expression of Progesterone Receptors A & B in Endometriosis Undergoing IVF Treatment: A Personalized Approach for Better IVF Success

Objective: Estrogen and Progesterone hormones play a pivotal role in the pathogenesis of Endometriosis. In the present study, relative quantification of PR-A and PR-B expression in eutopic endometrial tissue and endometriotic implants was done. By examining their expression profiles, we aim to gain deeper insights into the molecular mechanisms that can influence IVF success or failure in endometriotic patients. Our study also observed certain ART outcomes in both endometriosis and control groups, to evaluate the IVF success rate. Methods: This study was conducted at MHRT, a tertiary care center, and referral centers from other hospitals. The research encompassed an examination of 125 cases of infertility spanning from January 2019 to January 2023. Finally, biopsy samples were collected from 60 patients with endometriosis during laparoscopic procedures, while control biopsy samples were gathered from 18 patients who exhibited no signs of endometriosis lesions during the same laparoscopic procedures. The biopsy samples were then sent for Progesterone assay and later evaluated for ART outcomes. Results: It was observed from the results of our study that there was a significant downregulation of PR-A in stage III, and IV compared to Stage I and II. However, there was no significant change in PR-B expression in grades I and II, III and IV. Our study also observed certain ART outcomes in both endometriosis and control groups, revealing that women with endometriosis have lower chances of achieving pregnancy than women with other causes of infertility (95% CI, 22/60 vs. 12/18, p-value = 0.024). Conclusion: Our study results shed light on the complex role of progesterone isoforms in endometriosis. Progesterone receptor-A significant downregulation in severe cases challenges existing paradigms and opens new avenues for research.

ECM1 promotes migration and invasion in endometriosis

Extracellular matrix protein 1 (ECM1) is a glycoprotein that may be a key player in tumorigenesis and tumor progression. However, knowledge regarding the role of ECM1 in endometriosis (EM) is still lacking. Microarray analyses were performed to compare the mRNA expression patterns between paired EU tissues and ectopic endometrial (EC) tissues (n = 4) from EM patients. ECM1 expression was significantly increased in the eutopic endometrial (EU) tissues than paired EC tissues of endometriotic patients and normal endometrial (NE) tissues of controls without EM. Blocking ECM1 with siRNA attenuated the migration and invasion of hEM15A cells and modified the distribution of the F-actin cytoskeleton. We conducted microarray analyses and bioinformatics analyses to investigate the differentially expressed genes (DEGs) and related pathways regulated by ECM1. A total of 161 DEGs between the siECM1 and the negative control (siNC) treatments were identified, consisting of 79 downregulated genes and 82 upregulated genes. Enriched DEGs were associated with 9 gene ontology (GO) terms. Moreover, a protein-protein interaction (PPI) network was constructed for the hub genes and modules. Radixin (RDX) was the second most downregulated gene in the siECM1 group compared with the siNC group. ECM1 knockdown significantly decreased the expression of RDX, RhoC, ROCK1, N-cadherin and β-catenin but not ROCK2. ECM1 showed high tissue-specific expression in EU tissues from EM patients, and may contribute to the migration, invasion and reorganization of the F-actin cytoskeleton in eutopic endometrial stromal cells via the RhoC/ROCK1 signaling pathway in EM.

Evaluation of endometrioma size effect on ovarian reserve, embryo quality and pregnancy outcome after in vitro fertilization cycle; a cross-sectional study

IntroductionInvestigation of endometrioma size and its laterality on the quality of the embryo in patients with endometrioma compared to healthy subjects.Materials and methodsIn this retrospective and cross-sectional study, 70 patients with unilateral and bilateral endometrioma were recruited and compared with 70 age-matched infertile patients as the control group in terms of AMH before ovum pick-up, embryo quality as well as pregnancy outcome. Additionally, in the case group, we divided both unilateral (n = 32) and bilateral endometrioma patients (n = 38) into three groups based on the size of endometrioma. (1–3 cm, 3–6 cm, 6–10 cm)ResultsThere was no difference in terms of age, BMI, parity, and age of menarche between the case and control groups. Moreover, no significant difference was observed in the baseline level of AMH between the case 2.96 ± 2.72 ng/dl (0.21–11.3) and control 2.73 ± 2.39 (0.21–12.8) groups. (P = 0.59) There was also no significant difference concerning AMH level between unilateral 3.58 ± 3.20 ng/dl (0.21–12.8) and bilateral endometrioma 2.45 ± 2.14 (0.21 − 0.20) groups. In terms of the quality and number of embryos, there was no significant difference between the case and control groups. (P = 0.30) Although the AMH level decreased with the increase in endometrioma size, this difference was not statistically significant. (P = 0.07) There was no significant difference in terms of the embryo quality between the groups based on the size of endometrioma. (P = 0.77) In addition, no significant difference was observed between the case and control groups in the terms of birth weight and pregnancy complications, such as premature delivery, cesarean section rate, neonatal respiratory distress, jaundice, as well as hospitalization rate. Head circumference of the newborns was higher in the endometrioma group while their Apgar score was lower in the case compared to the control group.ConclusionThe presence of endometrioma by itself does not affect the main result of IVF procedures, including the number and quality of embryos and pregnancy outcome. Thus, IVF and embryo preservation and even pregnancy before surgery seem to be reasonable for endometriotic patients.

Comparative Study of Serum miRNA as A Biomarker for Non-Invasive Diagnosis in Suspected Versus Proven Cases of Endometriosis

Endometriosis is a chronic systemic inflammatory disorder characterized by endometrial tissue outside the uterine cavity in females. So far, the invasive laparoscopic method is the only gold standard diagnostic option for endometriosis. The study aims to develop a non-invasive diagnosis of endometriosis in patients presenting with one or more symptoms by analyzing the peripheral circulating miRNAs from the serum of the patients. A panel of miRNA 125b-5p, 342-3p, and Let-7b was developed to diagnose endometriosis. We performed the demographic profiling in 56 patients eliciting one or more symptoms of endometriosis without imaging evidence and compared them with 40 patients with the laparoscopically established endometriotic condition. Patients presenting with one or more of the clinical symptoms of endometriosis (n=56) served as the study group, and patients who were proven to be endometriotic by laparoscopy (n=40) served as the training (control) group. The fasting peripheral blood sample was collected, serum was separated, and cryo-preserved. qRT-PCR analysis of the selected miRNAs (miR 125b-5p, miR 342-3p, and Let-7b) was studied in both training (control) and study groups. The results were analyzed using a random forest (RF) approach machine learning algorithm and 10-fold cross-validation. Results indicate significant upregulation of miRNA125b-5p and miRNA 342-3p and downregulation of Let- 7b (p <0.001). Further, the samples derived from the study group with one or more symptoms of endometriosis and the proven endometriotic patients exhibited similar dysregulation of selected miRNAs (miR 125b-5p, miR 342-3p, and Let-7b). Based on our study, we propose the miRNAs panel consisting of miR 125b-5p, miR 342-3p, and Let-7b to be used as an early non-invasive diagnostic marker for endometriosis efficiently without the patients being subjected to invasive laparoscopy, which will reduce the time taken for diagnosis and commence the treatment earlier and prevent morbidity.

A novel therapeutic approach for endometriosis using adipose-derived stem cell-derived conditioned medium- A new hope for endometriotic patients in improving fertility.

Endometriosis is defined as the growth of endometrial glands and stromal cells in a heterotopic location with immune dysregulation. It usually leads to chronic pelvic pain and subfertility. Although various treatments are available, the recurrence rate remains high. Adipose tissue is an abundant source of multipotent mesenchymal adipose-derived stem cells (ADSCs). ADSCs display effects on not only tissue regeneration, but also immune regulation. Thus, the current study aims to test the effects of ADSCs on the growth of endometriosis. ADSCs isolated from lipoaspiration-generated adipose tissue and their conditioned medium (ADSC-CM) were subjected to quality validation, including karyotyping as well as growth promotion and sterility tests for microbial contamination under Good Tissue Practice and Good Manufacturing Practice regulations. An autologous endometriosis mouse model was established by suturing endometrial tissue to peritoneal wall followed by treating with DMEM/F12 medium, ADSC-CM, ADSCs or ADSC-CM+ADSCs for 28 days. The area of endometriotic cysts and the degree of pelvic adhesion were measured. ICAM-1, VEGF and caspase 3 expression was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. Moreover, the mice were allowed to mate and deliver. The pregnancy outcomes were recorded. The ADSC-CM was subjected to proteomics analysis with further data mining with Ingenuity Pathway Analysis (IPA). Both ADSC-CM and ADSCs passed quality validation. ADSC-CM reduced the area of endometriotic cysts. The inhibition by ADSC-CM was obliterated by adding ADSCs. The presence of ADSCs with or without ADSC-CM increased the peritoneal adhesion. ADSC-CM inhibited ICAM-1 and VEGF mRNA and protein expression, whereas the addition of ADSCs not only did not inhibit by itself, but also blocked the inhibition by ADSC-CM. The resorption rate was reduced by ADSC-CM. The number of live birth/dam and the survival rate of pup at 1 week-old were both increased by ADSC-CM in mice with endometriosis. IPA demonstrated that PTX3 was potentially critical for the inhibition of endometriosis by ADSC-CM due to its anti-inflammatory and antiangiogenic properties as well as its importance in implantation. ADSC-CM inhibited endometriosis development and improved pregnancy outcomes in mice. Potential translation to clinical treatment for human endometriosis is expected.

Review of the Potential Therapeutic Effects and Molecular Mechanisms of Resveratrol on Endometriosis.

Endometriosis is a hormone-dependent inflammatory disease characterized by the existence of endometrial tissues outside the uterine cavity. Pharmacotherapy and surgery are the current dominant management options for endometriosis. The greater incidence of recurrence and reoperation after surgical treatment as well as the adverse effects of medical approaches predispose patients to potential limitations for their long-term usage. Consequently, it is essential to explore novel supplementary and alternative drugs to ameliorate the therapeutic outcomes of endometriotic patients. Resveratrol is a phenolic compound that has attracted increasing interest from many researchers due to its pleiotropic biological activities. Here, we review the possible therapeutic efficacies and molecular mechanisms of resveratrol against endometriosis based on in vitro, animal, and clinical studies. The potential mechanisms of resveratrol include anti-proliferative, pro-apoptotic, anti-angiogenic, anti-oxidative stress, anti-invasive and anti-adhesive effects, thereby suggesting that resveratrol is a promising candidate for endometriosis. Because most studies have investigated the effectiveness of resveratrol on endometriosis via in vitro trials and/or experimental animal models, further high-quality clinical trials should be undertaken to comprehensively estimate the clinical application feasibility of resveratrol on endometriosis.

Targeting YAP1 ameliorates progesterone resistance in endometriosis

Does YAP1 inhibition alleviate progesterone resistance in endometriosis? YAP1 inhibition reduces progesterone resistance in vitro and in vivo. Progesterone resistance not only causes treatment failure for endometriosis but also inhibits eutopic endometrial cell proliferation, dysregulates decidualization, and reduces the success rates of pregnancy. Hippo/yes-associated protein 1 (YAP1) signaling pathway plays an important role in the pathogenesis of endometriosis. Paraffin-embedded tissues containing paired endometriotic and endometrial specimens (n = 42) and serum samples isolated from normal controls (n = 15) or endometriotic patients with (n = 25) or without (n = 21) prior dienogest treatment were analyzed. A mouse model of endometriosis was also used to evaluate the effects of YAP1 inhibition on progesterone resistance. Primary endometriotic and endometrial stromal cells treated with YAP1 inhibitor or miR-21 mimic/inhibitor were used for the in vitro studies including decidualization induction, chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation. Tissue specimens and serum from human and mouse were used for immunohistochemistry staining, exosome isolation, and microRNA (miRNA) quantification, respectively. Herein, we report, by using ChIP-PCR and RNA-IP, that YAP1 inhibits progesterone receptor (PGR) expression through upregulation of miR-21-5p. Upregulation of miR-21-5p not only reduces PGR expression but also inhibits endometrial stromal cell decidualization. Indeed, levels of YAP1 and miR-21-5p are inversely correlated with the level of PGR in human endometrial samples. In contrast, knockdown of YAP1 or treatment with verteporfin (VP), a YAP1 inhibitor, reduces miR-21-5p expression, thus leading to an increase in PGR expression in ectopic endometriotic stromal cells. In the mouse model of endometriosis, treatment with VP increases PGR expression and enhances decidualization. More importantly, VP synergistically increases the treatment effect of progestin in causing the regression of endometriotic lesions and improves the decidualization capability of the endometrium. Interestingly, treatment with dienogest, a synthetic progestin, reduces YAP1 and miR-21-5p expression in human cells and in the mouse model of endometriosis. Patients who received dienogest treatment for 6 months show a significant decrease in serum extracellular vesicle-associated miR-21-5p level. A public dataset (GSE51981) containing a large cohort of endometriotic tissues is available from the Gene Expression Omnibus (GEO). A large cohort of clinical samples is needed to verify the current diagnostic value of miR-21-5p in future studies. The reciprocal regulation of YAP1 and PGR suggests that combined YAP1 inhibitor and progestin may be a better therapeutic approach for treating endometriosis. This study was supported by the Ministry of Science and Technology, Taiwan (MOST-111-2636-B-006-012, MOST-111-2314-B-006-075-MY3, and MOST-106-2320-B-006-072-MY3). The authors have no conflict of interest to disclose.

Estetrol Increases Progesterone Genetic Response without Triggering Common Estrogenic Effects in Endometriotic Cell Lines and Primary Cultures

Estetrol (E4), a natural estrogen produced by the human fetal liver, is actively studied for menopause and breast cancer treatment. It has low side effects and preferential estrogen receptor alpha (ERα) affinity. There are no data about its effects on endometriosis, a common gynecological disease affecting 6-10% of cycling women, generating painful pelvic lesions and infertility. Current combined hormone treatment (progestins and estrogens) is safe and efficient; nevertheless, one-third of patients develop progesterone (P4) resistance and recurrence by reducing P4 receptors (PRs) levels. We aimed to compare E4 and 17β-estradiol (E2) effects using two human endometriotic cell lines (epithelial 11Z and stromal Hs832 cells) and primary cultures from endometriotic patients. We evaluated cell growth (MTS), migration (wound assay), hormone receptors levels (Western blot), and P4 response by PCR array. Compared to E2, E4 did not affect cell growth or migration but increased estrogen receptor alpha (ERα) and PRs, and reduced ERβ. Finally, the incubation with E4 improved the P4 gene response. In conclusion, E4 increased PRs levels and genetic response without inducing cell growth or migration. These results suggest that E4 might be useful for endometriosis treatment avoiding P4 resistance; however, evaluating its response in more complex models is required.

Correlations between Gut Microbial Composition, Pathophysiological and Surgical Aspects in Endometriosis: A Review of the Literature

Background and Objectives: Endometriosis is an estrogen-dependent, inflammatory, gynecological disorder represented by the migration of endometrial tissue outside the uterus. It can manifest through gynecological and gastrointestinal (GI) signs. Given the hormonal imbalances in endometriosis and the effect of microbiota on immune dysfunction, it has been thought that the human microbiome may play a role in its pathogenesis, acting differently before and after laparotomy. The aim of this review is to establish whether there is an interaction between endometriosis and gut microbial composition. Materials and Methods: We aimed to review available literature by systematically searching five databases: PubMed, EMBASE, Scopus, Cochrane Library, and ScienceDirect. We included records describing gut microbiota in the context of endometriosis-observing PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines-to recognize the presence of disease by the expression of bacterial taxa-based on 16S ribosomal RNA gene sequencing analysis. Results: Among 10 studies selected, there were four review articles and six clinical trials. The latter identified significant differences at a genus level in increased Prevotella, Blautia, and Bifidobacterium and decreased Paraprevotella, Ruminococcus, and Lachnospira (p < 0.05). In patients undergoing abdominal hysterectomy, Proteobacteria phylum increased from 34.36% before surgery to 54.04% after surgery (p < 0.05). Conclusions: Although scientific literature reports different characterizations of intestinal microbiota in endometriotic patients, further evidence is needed to develop new diagnostic-therapeutic strategies, for example, administration with probiotics before surgery.

Bioinformatics-based analysis of the roles of sex hormone receptors in endometriosis development.

Endometriosis is a hormone-dependent disease in women of reproductive age and seriously affects women's health. To analyze the involvement of sex hormone receptors in endometriosis development, we performed bioinformatics analysis using four datasets derived from the Gene Expression Omnibus (GEO) database, which may help us understand the mechanisms by which the sex hormones act in vivo in endometriosis patients. The enrichment analysis and protein-protein interaction (PPI) analysis of the differentially expressed genes (DEGs) revealed that there are different key genes and pathways involved in eutopic endometrium aberrations of endometriosis patients and endometriotic lesions, and sex hormone receptors, including androgen receptor (AR), progesterone receptor (PGR) and estrogen receptor 1 (ESR1), may play important roles in endometriosis development. Androgen receptor (AR), as the hub gene of endometrial aberrations in endometriotic patients, showed positive expression in the main cell types for endometriosis development, and its decreased expression in the endometrium of endometriotic patients was also confirmed by immunohistochemistry (IHC). The nomogram model established based on it displayed good predictive value.

Comparing the quality of life of endometriotic patients’ before and after treatment with normal and infertile patients based on the EHP30 questionnaire

ObjectiveThis study aimed to determine the quality of life (QOL), in patients with endometriosis ± infertility (B and C groups) and compare those to healthy women, and also infertile groups without endometriosis as a control groups (A and D), considering the fact that endometriosis and infertility reduces the quality of life in patients.MethodsThe present prospective comparative study was carried out between January 2018 and September 2020. A total of 400 women were included (100 women in each group). The participants filled in a validated questionnaire of quality of life, Endometriosis Health Profile-30 (EHP-30), and a visual analog scale of pain used, at the first visit, and 3 months after the medical or surgical treatment in the endometriosis group without infertility, additionally.ResultsThe majority of the patients were married, categorized in the middle-class of socio-economic state and housewives. They were of Persian descent. BMI was high in the infertile groups; however, the time of infertility was not different between the two groups of B and C (P = 0.054). The mean score of QOL was significantly lower in B, C, and D groups in comparison to the healthy women as the control group (A) (P < 0.001). Moreover, the infertile group (B), in comparison to endometriosis ± infertility groups (C and D), had the lowest mean score of QOL (P < 0.001). In each group, those who were older and had better educational level reported a better quality of life than other participants in that group. Social support plays a very important role in reducing the endometriosis related pain symptoms both before and after treatment. Three months after the treatment of endometriosis (D), a significant improvement was observed in all the aspects of QOL-related endometriosis. Nonetheless, the improvement of the quality of life in the surgical group was significantly higher than that in the medical treatment. The mean visual analog score of pain decreased from 62.22 ± 22.78, to 5.15 ± 2.73 following the surgical treatment (P < 0.001).ConclusionThe lowest quality of life belonged to the infertile group, followed by the endometriosis group. The quality of life of the endometriosis group improved after the treatment. Thus, endometriotic patients’ treatment in terms of improvement of quality of life should be considered by all professional health care teams.

The prognostic value of the cytological composition of the tear in the diagnosis of endometriosis

Purpose. To investigate the cell composition of the tear in healthy women and endometriosis patients in order to find the potential diagnostic criteria of endometriosis. Material and methods. Tear samples obtained from the lower fornix of the conjunctiva using a glass capillary tube were microscopically examined in 100 females aged 18 to 48: 60 healthy women, who made up the control group, and 40 patients with endometriosis. Results. The share of women with endometriosis who were found to have erythrocytes was 32.5 %, which was significantly higher as compared with the control group, where it was 25.0%. 69.2 % of women with endometriosis showed haemolacria in the follicular phase of the menstrual cycle, which significantly exceeded the percentage of women with erythrocytes in the tear (30.8 %) who had them in the luteal phase. Epitheliocytes were found in the tear of 97.5 % of patients with endometriosis, versus 75.0 % of healthy women. In all 100 % of such patients, these cells were found in the follicular phase of the menstrual cycle, compared with 74.3 % of healthy women who showed these cells in the same phase of the cycle. The share of women with endometriosis showing the ++ quantity of epitheliocytes in the tear was 56.4 %, which was significantly higher than the 20 % of the control group subject with the same level of epitheliocytes. It is to be noted that group accumulations of epitheliocytes were found in 15 % of endometriotic patients, which was significantly higher than in healthy women, of whom only 8.3 % had such accumulations. 90 % of endometriosis group cases revealed glandular cuboidal epithelial cells in the tear, which morphologically resembling endometrial cells. Conclusion. The revealed properties of the cytological composition of the tear of patients with endometriosis reflected particular links in its etiopathogenesis, which suggests that the study of tear parameters can help predict the incidence of the "endometrioid disease" and develop pathogenetically oriented treatment methods.

Stomatin-like Protein-2 Promotes Aggregation, Colonization and Migration of Endometriotic Cells.

Endometriosis is a chronic gynecological disease in women of childbearing age, which leads to infertility with risk of endometrial and ovarian cancer. The pathogenesis of endometriosis is poorly understood, and cure/treatment for it is not available, except for symptomatic treatment. The recurrence rate of endometriosis is high. SLP-2 is an inner mitochondrial membrane protein whose participation has been explained in cases of endometrial stromal cell growth, differentiation and migration, but its role in endometriosis is yet to be understood. Previous studies have found altered expression of stomatin-like protein 2 (SLP-2) in the serum of endometriotic patients. Therefore, we have studied the possible role of SLP-2 in the development of endometriosis. We found the ubiquitous and high expression of SLP-2 in the endometriotic tissue of both human endometriosis patientsand rat endometriosis model. SLP-2 is seen in the glandular epithelial cells and stromal cells in the eutopic/normal or non-endometriosis group endometrium from human subjects. Finding high expression levels of SLP-2 in endometriotic tissue and ovarian cystic cells derived from endometriosis patients, we explored the possible role of SLP-2 in the cell aggregation, colonization, migration, and invasion in the human endometriotic cells associated with the progression of the endometriosis. Transient silencing of SLP-2 by its siRNA hinders endometriotic cells, aggregation, migration, and invasion into the extracellular matrix, which confirms SLP-2 involvement in endometriotic disease onset and progression. This study unravels the ubiquitous expression of SLP-2 in the human ectopic endometrial tissue and its role in the endometriotic cell migration, colonization, aggregation, and invasion leading to endometriosis progression.

Effect of active vitamin D on proliferation, cell cycle and apoptosis in endometriotic stromal cells

Research questionWhat is the effect of vitamin D3 (1,25(OH)2D3) on proliferation, cell cycle and apoptosis of endometrial stromal cells (ESC) in endometriotic patients? DesignESC isolated from 10 women with endometriosis and 10 healthy controls were treated with 1,25(OH)2D3. The proliferation of control endometrial stromal cells (CESC), eutopic endometrial stromal cells (EuESC) and ectopic endometrial stromal cells (EESC) was analysed 72 h after the treatment using methyl thiazolyl tetrazolium assay. Propidium iodide staining and flow cytometry were used to determine the cell cycle distribution in ESC. Annexin V/propidium iodide double staining was used to evaluate apoptosis in ESC. ResultsIn the presence of oestrogen, 1,25(OH)2D3 treatment inhibited the proliferation of ESC from all three origins (P = 0.009 for CESC, P = 0.005 for EuESC and P < 0.001 for EESC). The percentage of S phase cells in EESC was higher than in EuESC and CESC (P = 0.002 and P = 0.001, respectively). The percentage of S phase cells in EuESC was higher than in CESC (P = 0.005). The percentage of G1 phase cells in EESC was lower than that of EuESC and CESC (P = 0.003 and P = 0.002, respectively) and the percentage of G1 phase cells in EuESC was lower than that of CESC (P = 0.007). Moreover, 1,25(OH)2D3 inhibited cell cycle regardless of cell type (P = 0.002 in EESC, P = 0.001 in EuESC and P = 0.014 in CESC), but in the absence of oestrogen, inhibited cell cycle only in EuESC (P = 0.012). ConclusionsAlthough 1,25(OH)2D3 increased apoptotic and necrotic cells and decreased live cells in the EuESC and EESC, it did not affect apoptosis in CESC and only increased necrotic cells. These findings indicate that 1,25(OH)2D3 potentially has a growth-inhibiting and pro-apoptotic effect on ESC from endometriotic patients.

Changes in MCP-1, HGF, and IGF-1 expression in endometrial stromal cells, PBMCs, and PFMCs of endometriotic women following 1,25(OH)2D3 treatment.

1,25(OH)2D3 has anti-inflammatory and growth inhibitory effects. Our study explored the effect of 1,25(OH)2D3 treatment on the expression of monocyte chemotactic protein-1 (MCP-1), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) by peripheral blood mononuclear cells (PBMCs), peritoneal fluid mononuclear cells (PFMCs), endometrial stromal cells (ESCs), and its effect on the proliferation of PBMCs and PFMCs of patients with endometriosis compared with controls. PBMCs, PFMCs, and ESCs were obtained from 10 endometriosis patients and 10 non-endometriotic individuals. After treating cells with 0.1μM of 1,25(OH)2D3 for 6, 24, and 48 h, the gene and protein expression of mentioned factors were evaluated by real-time PCR and ELISA methods, respectively. 1,25(OH)2D3 treatment significantly reduced the protein expression of MCP-1, HGF, and IGF-1 in PBMCs and PFMCs of endometriotic patients at 48 h (p&lt; 0.05-&lt;0.01). Also, this treatment significantly reduced MCP-1, HGF, and IGF-1 gene and/or protein expression in EESCs and EuESCs at 24 and 48 h (p&lt; 0.05-&lt;0.01). 1,25(OH)2D3 treatment also reduced the proliferation of PBMCs and PFMCs of endometriotic patients compared with controls (p &lt; 0.01). 1,25(OH)2D3 can be considered as a potentially effective agent in the prevention and treatment of endometriosis along with other therapies.

Spatiotemporal expression pattern of Progesterone Receptor Component (PGRMC) 1 in endometrium from patients with or without endometriosis or adenomyosis

The endometrium plays a crucial role in reproduction and, in humans, is cyclically remodeled under hormonal control. Estradiol favors tissue proliferation whereas progesterone inhibits tissue growth and induces morphological changes. Endometriosis is often associated with fertility issues and with exacerbated estrogen and reduced progesterone concentration or response in the eutopic endometrium. However, underlying mechanisms remain unclear. Progesterone Receptor Membrane Component (PGRMC) 1 is a protein able to modulate progesterone response and its murine knockout reduced fertility. However, the precise spatiotemporal pattern of PGRMC1 expression in the human endometrium is still poorly characterized. We investigated variations of eutopic endometrial PGRMC1 expression by combining RT-qPCR, immunofluorescence and in situ hybridization. We found that PGRMC1 expression progressively increases during the proliferative phase and decreases during the secretory phase. However, immunolabeling and identification of mRNA-containing cells were regularly heterogeneous in samples, according to tissue depth, with a gradient extending from the surface epithelium towards the basalis. There was no significant difference in PGRMC1 mRNA amounts between patients with or without endometriosis or adenomyosis, for any phase of the menstrual cycle, but cells with strong or moderate PGRMC1 immunolabeling were reduced during the proliferative phase in endometriotic patients. In conclusion, although the cyclical variation of PGRMC1 expression globally follows fluctuation of ovarian steroids, further work is required to precisely characterize hormonal control and identify the additional levels of regulation responsible for local adjustment of PGRMC1 concentration. This is particularly important in the light of recent studies emphasizing the correlation between adequate PGRMC1 amounts and fertility.

O-032 Primary human endothelial and stromal cells from the uterine endometrium co-cultured in vitro in a 3D-system as a model to study the physiopathology of endometriosis

Abstract Study question Is an uterine endothelial-stromal cell 3D-system able to respond to inflammatory/immune factors presented in patient’s endometriotic serum and can this be reversible by hormonal treatment? Summary answer The endothelial-stromal cells system is responsive to the serum from women with endometriosis and its cytokine profile may be reverse with hormonal treatment. What is known already Endometriosis’s declined fertility is mainly attributed to poor oocyte quality, inhibition of ovulation, an anatomical commitment of tubes and uterus, and loss of endometrial receptivity during the implantation window. Changes in the inflammatory/immune profile in pelvic and peripheral blood also suggest a possible interference of several cytokines playing a role in women's reduced fertility. Three-dimensional (3D) cell cultures open up new study possibilities, by guaranteeing interactions between spatially organized tissues mimicking the natural microenvironment and can significantly contribute to obtaining essential data for understanding endometrial physiology and its associated diseases. Study design, size, duration This is a prospective cohort study with oocyte donation women from a private IVF center and patients under endometriosis treatment in an University-affiliated gynecology service. Endometrium biopsy (n = 9) and non-endometriotic serum blood (n = 15) were collected from oocyte donors in the same day of oocyte picked (antagonist protocol), before ovary puncture and serum blood samples were collected from patients diagnostic with endometriosis (n = 15). Samples were collected between Jan/2016 and May/2017 after signing the informed consent form. Participants/materials, setting, methods Endometriotic serum samples were obtained from patients with (n = 10) or without (n = 5) estrogen/progestin therapy. Tissue biopsies were digested and submitted to magnetic microbeads. Endothelial and stromal cells layers were added one-by-one to a mixture of extracellular matrix components. The 3D-system received endometriotic or control serum for additional 48h. Supernatants were excluded and the cells homogenized for cytokine evaluation through cytometric bead array. The ANOVA-Tukey’s test were used for statistical analysis, p &amp;lt; 0.05 were considered significant. Main results and the role of chance Oocyte donors were 24,2 ± 3,41 years old and had a body mass index (BMI) of 22,7 ± 1,38. Patients with endometriosis were 34,0 ± 3,93 years old (p = 0,21) and had a BMI of 25,4 ± 4,13 (p = 0.02). The morphology assessed by ultrastructural analysis of the 3D-system showed the presence of cells embedded in an abundant extracellular matrix, with no directional organization and with morphology compatible with fibrocytes and fibroblasts. The cells showed characteristics of viable cells with loose chromatin nuclei, evident nucleoli, and well-defined organelles. Eventually, mitosis were observed. Specific presence or absence of stromal and endothelial cells markers (cytokeratin, vimentin, IGFBP1, von Willebrand factor) were confirmed by immunofluorescence and flow cytometry. Cell viability were assessed by Fixable Cell Viability 510 dye staining. Changes in the expression profile of the cytokines TNF-alpha (p &amp;lt; 0.0001), IL-10 (p &amp;lt; 0.001) and IL-2 (p &amp;lt; 0.001) were detected in the endothelial-stromal cells treated with the serum of endometriotic patients in comparison with hormonally-treated and non-endometriotic groups. These results suggest amelioration of the immune response by endometrial cells when submitted to a serum environment under hormonal treatment. Serum of endometriotic patients (hormonally treated and non-treated) also increased the levels of IL-6 and IL-8 in the co-cultured cells. Limitations, reasons for caution Endometrial biopsies were collected in the initial secretory phase of young oocyte donor women, which may not reflect to the eutopic endometrial response in patients with endometriosis. The reduced size of patients cohort may also increase the risks of bias in cytokine analysis. Wider implications of the findings The possibility of endometrial cells, even in the absence of leukocytes, to become a protagonist in the expression of cytokines and the ability of uterine cells to respond and contribute to a systemic inflammatory/immunomodulating environment with cytokine production may reflect on the pathophysiology of endometriosis and uterine reproductive functions. Trial registration number Not Applicable