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Background: Endometriosis is a chronic disease mostly affecting women at reproductive age. Some evidence suggests that surgery of ovarian endometrioma may have a detrimental effect on ovarian reserve. The aim of this study was to evaluate the changes of serum antimullerian hormone (AMH) levels in patients with endometrioma after cystectomy. Methods: A prospective study was performed at Nikan hospital on 58 patients with endometrioma who underwent laparoscopic cystectomy. Of them, 30 had unilateral endometrioma and 28 had bilateral endometrioma. Complete excision was done, pelvic endometriosis implants as well as deep infiltrative endometriosis was resected. Sutures were made for the closure of ovarian parenchyma and bleeding control. We did not use any hot energy devices such as cautery on ovaries for ablation, coagulation or resection of endometrioma. Serum AMH levels were measured preoperatively, 3, 9, and 15 months postoperatively. Results: Serum AMH levels decreased significantly from the preoperative sample (2.98 ± 2.47 ng/ml) to 3 months after laparoscopy (1.07 ± 1.06 ng/ml), then gradually increased 9 months (1.47 ± 1.16 ng/ml) and 15 months (1.95 ± 1.85 ng/ml) after surgery, without returning to the preoperative levels during the follow-up time of study. Conclusion: There is a fluctuation pattern in AMH levels from preoperative to 15-month follow-up after endometrioma surgery using only suture for ovarian hemostasis. Firstly, there is decline in AMH level 3 months after surgery, then an increasing trend was observed gradually up to 15 months after surgery. Controlled studies are needed to compare the effects of various cystectomy methods on the ovarian reserve after endometrioma surgery

Background Endometriosis is a chronic, painful, and inflammatory disease characterized by extra-uterine growth of endometrial tissues. Increased angiogenesis and resistance to apoptosis have been suggested to be involved in pathogenesis and development of endometriosis. The objective of this study was to examine apoptosis potential and angiogenesis contribution of eutopic (EuESCs) and ectopic (EESCs) endometrial stromal cells in patients with endometriosis compared to endometrial stromal cells from non-endometriotic controls (CESCs). Methods Stromal cells were isolated by enzymatic digestion of ectopic (n = 11) and eutopic (n = 17) endometrial tissues from laparoscopically-confirmed endometriotic patients. Endometrial stromal cells of 15 non-endometriotic patients served as control. Following cell characterization by immunofluorescent staining and flow cytometry using a panel of antibodies, the total RNA was isolated from the cultured cells, and analyzed for the expression of genes involved in apoptosis (Bcl-2, Bcl-xL, Bax, and caspase-3) and angiogenesis [vascular endothelial growth factor-A (VEGF-A) and hepatocyte growth factor (HGF)] by Real-time PCR. Results Significantly higher gene expression levels of Bcl-2 and Bcl-xL were found in EESCs compared with EuESCs and CESCs (p p p p p p < 0.05). Conclusions These findings suggest reduced propensity to apoptosis and increased angiogenesis potential of EESCs, which may be involved in pathogenesis of endometriosis.

Endometriosis is one of the most common chronic gynecological disorders affecting women at reproductive age. Dysregulation of immune cells, including regulatory T (Treg) cells has contributed to the growth of ectopic lesion in patients with endometriosis.The present study investigated the frequency of Tregs in peripheral blood and the expression of Foxp3 in eutopic and ectopic endometriotic tissues in women with and without endometriosis.Peripheral blood mononuclear cells (PBMCs) and eutopic and ectopic endometriotic tissues were obtained from 23 endometriotic and 20 non-endometriotic control women. The frequency of Treg cells in PBMCs was measured using flowcytometry and the expression of Foxp3 in eutopic and ectopic endometriotic tissues was determined by real-time PCR, western blotting and immunohistochemistry.The frequency of circulating Tregs was significantly higher in endometriotic patients compared with non-endometriotic controls (P0.01). The mRNA and protein expression of Foxp3 in eutopic and ectopic endometriotic tissues had no significant differences between the two study groups.Higher frequency of circulating Tregs in patients with endometriosis compared with controls may be considered as a compensatory mechanism to regulate the inflammatory condition in this disease.

Objective To evaluate the result of laparoscopic endometrioma excision in fertility outcome of advanced endometriosis patients. Study design The study was designated as historical cohort, in a private referral center of advance laparoscopy. 111 infertile patients, diagnosed as endometriosis, were divided in two groups: DIE (deep infiltrative endometriosis) and endometrioma (case group), and patients with only DIE (without endometrioma ((control group). All patients underwent global laparoscopic resection of DIE lesion (both groups) and laparoscopic excisional cystectomy of endometrioma (case groups). Patients were followed for fertility outcomes and data were analyzed by Kaplan–Meier test and COX regression using SPSS software. Results After adjusting covariates, the Kaplan–Meier analysis of cumulative pregnancy rates (CPR) did not show any statistical significance between cases (35.6%) and controls (39.5%) (Log-rank P -value = 0.959). The COX regression analysis of covariates showed there is no significant relationship between cystectomy and fertility outcome. It showed statistical significance effect of age (hazard ratio [HR] = 0.772), years of infertility (HR = 0.224), and previous endometrioma surgery (HR = 0.180), on fertility chance. Conclusion In advanced endometriosis with DIE and infertility, fine excision and stripping of the endometrioma along with radical resection of DIE improves fecundity without any significant adverse effect in comparison with patients with intact ovaries.

Retrograde flow of menstrual blood cells during menstruation is considered as the dominant theory for the development of endometriosis. Moreover, current evidence suggests that endometrial-derived stem cells are key players in the pathogenesis of endometriosis. In particular, endometrial stromal stem cells have been suggested to be involved in the pathogenesis of this disease. Here, we aimed to use menstrual blood, as a novel source of endometrial stem cells, to investigate whether stromal stem cells from endometriosis (E-MenSCs) and non-endometriosis (NE-MenSCs) women differed regarding their morphology, CD marker expression pattern, proliferation, invasion and adhesion capacities and their ability to express certain immunomodulatory molecules. E-MenSCs were morphologically different from NE-MenSCs and showed higher expression of CD9, CD10 and CD29. Furthermore, E-MenSCs had higher proliferation and invasion potentials compared with NE-MenSCs. The amount of indoleamine 2,3-dioxygenase-1 (IDO1) and cyclooxygenase-2 (COX-2) in E-MenSCs co-cultured with allogenic peripheral blood mononuclear cells (PBMCs) was shown to be higher both at the gene and protein levels, and higher IDO1 activity was detected in the endometriosis group. However, NE-MenSCs revealed increased concentrations of forkhead transcription factor-3 (FOXP3) when compared with E-MenSCs. Nonetheless, interferon (IFN)-γ, Interleukin (IL)-10 and monocyte chemoattractant protein-1 (MCP-1) levels were higher in the supernatant of E-MenSCs-PBMC co-cultures. Here, we showed that there are inherent differences between E-MenSCs and NE-MenSCs. These findings propose the key role MenSCs could play in the pathogenesis of endometriosis and further support the retrograde and stem cell theories of endometriosis. Hence, considering its renewable and easily available nature, menstrual blood could be viewed as a reliable and inexpensive material for studies addressing the cellular and molecular aspects of endometriosis.

Objective To study immunophenotype, differential proliferation capacity, invasiveness, adhesion, and cytokine production in ectopic and eutopic endometrial stromal cells (EESCs and EuESCs) from patients with endometriosis. Design In vitro study. Setting Academic research center. Patient(s) Patients with ovarian endometriosis (endometrioma) and nonendometriotic controls. Intervention(s) None. Main Outcome Measure(s) EESCs and EuESCs from 25 patients with endometrioma and ESCs from 20 nonendometriotic controls (CESCs) were isolated, and their immunophenotype, proliferation, invasion, adhesion, and cytokine production were assessed and compared. Result(s) Isolated ESCs from all three sources expressed markers specific for cells of mesenchymal origin but were negative for hematopoietic markers. EESCs exhibited a significantly lower proliferation rate in fibronectin-coated plates and less invasive capacity compared with CESCs or EuESCs. Among all stromal cell groups studied, EuESCs showed the highest invasive behavior. EESCs adhered more firmly to extracellular matrix than EuESCs or CESCs in all time intervals examined. The levels of interleukin (IL) -6 and IL-8 production by EESCs were significantly higher compared with those of EuESCs or CESCs. Conclusion(s) The results of the present study demonstrated that retrograde menstruation alone does not account for the pathogenesis of endometriosis as eutopic and ectopic counterparts of ESCs from patients with endometriosis exhibit differential invasive, adhesive, and proliferative behavior.

Aim: Menstrual blood stromal stem cells (MBSCs) have been demonstrated to exhibit stem cell properties such as the capability for self-renewal and multipotency, allowing for multilineage differentiation. In addition, this cell type has various immunomodulatory effects. In this study, we examined the potential effect of MBSCs on proliferation of peripheral blood mononuclear cells (PBMCs) in allogeneic mixed lymphocyte reaction (MLR). Materials and Methods: Menstrual blood was collected from healthy donors after menstrual blood flow initiated and its mononuclear cell fraction was separated. Cells were subsequently cultured and adherent cells were allowed to propagate and used as stem cells. Flowcytometric immunophenotyping was performed using a panel of monoclonal antibodies including CD44, CD45, CD34, CD9, CD29, CD10, CD38, CD105, CD73, CD133, STRO-1 and Oct-4A. For functional analysis, PBMCs were co-cultured with MBSCs, collected after 4 days and added to allogeneic PBMCs. 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was carried out to evaluate cell proliferation. Results: MBSCs showed surface and intracellular markers of mesenchymal stem cells with the exception of the high expression of Oct-4A. MBSCs affected the proliferative response of PBMC in a dose-dependent manner. At ratio of 1:1 to 1:2, MBSCs inhibited, while at lower ratios (1:32 to 1:64) stimulated the proliferative capacity of allogeneic PBMCs. Conclusion: According to the present study, MBSCs exert their immunoregulatory effects on allogeneic PBMCs in a dose-dependent manner. This finding can be considered as a valuable point in future cell therapy strategies, when this cell population is used.

Problem We aimed to evaluate modulatory effects of vitamin D3 on endometriosis-related features of endometriotic stromal cells. Method of study The effect of vitamin D3 on adhesion, invasion, proliferation, apoptosis, cytokine production, and angiogenesis potential of the eutopic (EuESCs), ectopic (EESCs), and control (CESCs) stromal cells from 25 women with and 20 women without endometriosis was investigated. Results In all groups, vitamin D3 significantly increased cell adhesion (P = 0.0013–0.042), while decreased invasion (P = 0.026–0.031) and proliferation (P = 0.0013–0.039) of EESCs and EuESCs. Such treatment also resulted in a significant decrease in IL-6 production by EESCs (P = 0.039), but had no significant effect on the IL-8 production. This vitamin also caused significant decrease in Bcl-2 gene expression by EuESCs (P = 0.04) and Bcl-xL by EESCs (P = 0.044–0.009). In addition, vitamin D3 treatment reduced VEGF-A gene expression by EESCs (P = 0.046–0.009). Conclusion Based on substantial favourable in vitro effects of vitamin D3 in endometriosis-related features of human endometriotic stromal cells, further investigations on therapeutic potential of this hormone in endometriosis are warranted.

Menstrual blood stromal stem cells (MBSCs) have been demonstrated to exhibit stem cell properties such as the capability for self-renewal and multipotency, allowing for multilineage differentiation. In addition, this cell type has various immunomodulatory effects. In this study, we examined the potential effect of MBSCs on proliferation of peripheral blood mononuclear cells (PBMCs) in allogeneic mixed lymphocyte reaction (MLR).Menstrual blood was collected from healthy donors after menstrual blood flow initiated and its mononuclear cell fraction was separated. Cells were subsequently cultured and adherent cells were allowed to propagate and used as stem cells. Flowcytometric immunophenotyping was performed using a panel of monoclonal antibodies including CD44, CD45, CD34, CD9, CD29, CD10, CD38, CD105, CD73, CD133, STRO-1 and Oct-4A. For functional analysis, PBMCs were co-cultured with MBSCs, collected after 4 days and added to allogeneic PBMCs. 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was carried out to evaluate cell proliferation.MBSCs showed surface and intracellular markers of mesenchymal stem cells with the exception of the high expression of Oct-4A. MBSCs affected the proliferative response of PBMC in a dose-dependent manner. At ratio of 1:1 to 1:2, MBSCs inhibited, while at lower ratios (1:32 to 1:64) stimulated the proliferative capacity of allogeneic PBMCs.According to the present study, MBSCs exert their immunoregulatory effects on allogeneic PBMCs in a dose-dependent manner. This finding can be considered as a valuable point in future cell therapy strategies, when this cell population is used.