Endometrial Pathology Research Articles

Autophagy Involvement in Non-Neoplastic and Neoplastic Endometrial Pathology: The State of the Art with a Focus on Carcinoma

Autophagy is a cellular process crucial for maintaining homeostasis by degrading damaged proteins and organelles. It is stimulated in response to stress, recycling nutrients and generating energy for cell survival. In normal endometrium, it suppresses tumorigenesis by preventing toxic accumulation and maintaining cellular homeostasis. It is involved in the cyclic remodelling of the endometrium during the menstrual cycle and contributes to decidualisation for successful pregnancy. Such a process is regulated by various signalling pathways, including PI3K/AKT/mTOR, AMPK/mTOR, and p53. Dysregulation of autophagy has been associated with benign conditions like endometriosis and endometrial hyperplasia but also with malignant neoplasms such as endometrial carcinoma. In fact, it has emerged as a crucial player in endometrial carcinoma biology, exhibiting a dual role in both tumour suppression and tumour promotion, providing nutrients during metabolic stress and allowing cancer cell survival. It also regulates cancer stem cells, metastasis and therapy resistance. Targeting autophagy is therefore a promising therapeutic strategy in endometrial carcinoma and potential for overcoming resistance to standard treatments. The aim of this review is to delve into the intricate details of autophagy’s role in endometrial pathology, exploring its mechanisms, signalling pathways and potential therapeutic implications.

Endometrial pathology among premenopausal patients seeking emergency care for abnormal uterine bleeding: High risk of endometrial malignancy and premalignancy

Endometrial pathology among premenopausal patients seeking emergency care for abnormal uterine bleeding: High risk of endometrial malignancy and premalignancy

Screening for endometrial disease: Analyzing risk factors for abnormal endometrial pathology in a high-risk population

Screening for endometrial disease: Analyzing risk factors for abnormal endometrial pathology in a high-risk population

The role of herpesviruses in endometrial pathology: frequency of herpesvirus DNA detection in the endometrium, cervix and vagina in patients with chronic endometritis and endometrial hyperplasia

Introduction. Chronic endometritis (CE) is reported in 12% of patients with primary infertility. Herpesviruses are considered as one of the causes of CE.The aim of the study. To evaluate the frequency of herpesviruses DNA detection in endometrium, cervix and vagina in patients with chronic endometritis and endometrial hyperplasia.Materials and methods. 167 women of reproductive age were included in the study. Samples from the posterolateral wall of the vagina, cervix and endometrium were taken from each patient. The obtained samples were analyzed by real-time PCR for the presence of Herpes simplex viruses type 1 and 2 (HPV 1,2), Epstein-Barr virus (EBV), Cytomegalovirus (CMV) and Human herpesvirus type 6 (HHV6). Endometrial tissue was subjected to morphologic examination.Results. Depending on the morphology of endometrial biopsy, all patients were divided into three groups: with CE (N=53), with endometrial hyperplasia (EH, N=63), with normal endometrium (N=51). There were no statistically significant differences in the frequency of herpesvirus detection in endometrial, cervical, and vaginal samples. In endometrial samples, EBV was detected in 1 (1.9%) patient with CE (1.9%) and 1 (1.6%) patient with EH, HPV1,2 — in 1 (2%) woman with normal endometrial histology; DNA of other herpesviruses was not detected.Conclusions. The relationship between herpesvirus type, positive locus and the presence of endometrial pathology has not been established. The frequency of herpesvirus DNA detection in endometrial, cervical, and vaginal samples was not significantly different between the groups of women with CE, EH, and normal endometrial histology. Only 2 (1.7%) of endometrial samples with pathologic morphology were positive for DNA of herpesviruses, in both cases it was EBV.

Breast and endometrial safety of micronised progesterone versus norethisterone acetate in menopausal hormone therapy (PROBES): study protocol of a double-blind randomised controlled trial

IntroductionData suggest that micronised progesterone (mP) in menopausal hormone therapy is safer for the breast than synthetic progestins, while protection of the endometrium appears to be less effective. However, comparative...

Structural and functional features of the endometrium in chronic endometritis and implantation failure in IVF cycles

BACKGROUND: Implantation failure of the endometrium in women with chronic endometritis remains a subject of discussion, despite certain progress in the diagnosis and treatment of this pathology. It has been proven that chronic endometritis is a predictor of endometrial dysfunction, which contributes to the disruption of the morphogenesis of the endometrium necessary for its implantation capacity. Implantation disorders associated with chronic endometritis are related to pathology of the receptor profile and vascular bed, as well as dysregulation of proliferative and apoptotic mechanisms. AIM: The aim of this study was to evaluate the expression of estrogen and progesterone receptors, endothelial marker and apoptosis-inducing factor in the endometrium of patients with chronic endometritis and ineffective embryo transfer in IVF protocols. MATERIALS AND METHODS: This study enrolled 50 patients with ineffective embryo transfers in IVF protocols and 25 women in the control group with positive reproductive outcomes. All patients underwent a comprehensive morphological study of the endometrium including histological examination, immunohistochemical study with assessment of the receptor profile [estrogen (ERα) and progesterone (PR) receptor expression], chronic endometritis (CD8+, CD4+, CD20+, CD138+), apoptosis [antibodies to apoptosis-inducing factor (anti-AIF [E20])], and angiogenesis (CD34+), with microscopic, morphometric and statistical methods used as well. RESULTS: Endometrial implantation failure is associated with a history of urogenital infection, benign cervical diseases, high incidence of endometrial pathology and reproductive loss. It is characterized by the presence of follicle-like mononuclear infiltrates in the endometrium, fibroplastic changes in the stromal component, vascular sclerosis, abnormal increase in the pro-inflammatory cell count, multifocal decrease in estrogen (ERα) and progesterone (PR) receptor expression, as well as activation of pathological neoangiogenesis and apoptosis in the stroma. CONCLUSIONS: Chronic endometritis leads to disruption of the structural and functional characteristics of the endometrium, stimulates pathological neoangiogenesis and dysregulates proliferative and apoptotic mechanisms, which forms a vicious circle of endometrial implantation failure and inefficiency of embryo transfer in IVF protocols.

8314 Masculinizing Hormone Therapy and Gynecological Neoplasm: a Single Institution Cohort

Abstract Disclosure: S. Korpaisarn: None. T. Ngaothepphurktharam: None. Y. Tingthanatikul: None. Background: Gynecological neoplasm/malignancy is one of the concerned areas among transgender men on masculinizing hormone therapy (MHT). Long-term MHT and gynecological neoplasm/malignancy remain unclear. Methods: A retrospective cohort chart review was done on all transgender men (TM) who underwent hysterectomy with or without oophorectomy between January 2014 and October 2022. Intraoperative findings and pathological results of genital organs, including the cervix, endometrium, myometrium, and fallopian tubes, were extracted. The pathological results were classified as benign, precancerous, and malignant. Descriptive statistical analysis was used to demonstrate the prevalence of gynecological neoplasm and malignancy. Results: A total of 30 TM underwent hysterectomy with oophorectomy. Twenty-eight (93%) had bilateral oophorectomy, and 7% had unilateral oophorectomy due to gynecological conditions. The mean age when TM underwent surgery was 32.1 ± 5.1 years, and the mean age of gender-affirming hormone initiation was 28.7± 5.1 years. The median duration of MHT before gynecological surgery was 32 months (IQR 22-59). Most TM (73%) used testosterone enanthate, 17% testosterone undecanoate, and 2 transgender men did not start MHT before genital surgery. All TM who used GHT became amenorrheic before surgery. The median preoperative testosterone level was 561.5 ng/dL (IQR 776-1189). Intraoperative findings revealed that most TM (86.7%) had unremarkable uterus size, and 13% showed enlarged uterus size. In the latter, all were from myoma uteri. Most intraoperative findings of ovaries showed 93% unremarkable. One TM had intraoperative polycystic ovarian morphology, and one showed a unilateral ovarian cyst. Cervical pathological reports showed unremarkable results in 67%. Five TM (17%) had chronic cervical inflammation. There was 1 TM (3%) who showed low-grade squamous intraepithelial lesion (LSIL) with squamous metaplasia. Regarding endometrial pathology, the mean endometrial thickness was 0.13 ± 0.08 cm. Most endometrial reports (66%) were in a proliferative phase, 27% were in an atrophic or inactive endometrium, and 7% were in the secretory phase. Regarding pathological reports of ovaries, 80% had cystic ovaries. None of the TM showed precancerous or malignant conditions of endometrium, ovaries, and fallopian tubes. PAP smear was available in 27 TM; all were negative for intraepithelial lesions or malignancy. Conclusion: Among a cohort of TM who received MHT for 32 months, there is no evidence of precancerous or malignant pathology of the endometrium, ovaries, and fallopian tubes. Most of them had thin endometrium with cystic ovaries. One TM showed LSIL with squamous metaplasia of the cervix. Healthcare providers should follow local gynecological cancer screening guidelines developed for cisgender women in their clinical practice. Presentation: 6/2/2024

ECMTrans-net: Multi-Class Segmentation Network Based on Tumor Tissue in Endometrial Cancer Pathology Images

Endometrial cancer has the second highest incidence of malignant tumors in the female reproductive system, and accurate and efficient endometrial cancer pathology image analysis is one of the important research components of computer-aided diagnosis. However, endometrial cancer pathologic images have the challenges of smaller solid tumors, lesion areas varying in morphology, and difficulty distinguishing solid and non-solid tumors, which would impact the accuracy of subsequent pathological analyses. Therefore, an Endometrial Cancer Multi-class Transformer Network (ECMTrans-net) is proposed to improve the segmentation accuracy of endometrial cancer pathology images. On the one hand, an ECM-Attention is proposed, which can sequentially infer attention maps along three separate dimensions: channel, local spatial, and global spatial, and multiply the attention maps and the input feature map for adaptive feature refinement, solving the problems of the small size of solid tumors and similar characteristics of solid tumors to non-solid tumors and further improving the accuracy of segmentation of solid tumors. On the other hand, an ECM-Transformer is proposed, which can fuse multi-class feature information and dynamically adjust the receptive field, solving the issue of complex tumor features. Experiments on the solid tumor endometrial cancer pathological (ST-ECP) dataset show that the ECMTrans-net performs superior to state-of-the-art image segmentation methods, and the average values of Accuracy, MIoU, Precision, and Dice were 0.952, 0.927, 0.931 and 0.901, respectively.

A Deeper Look at Office Hysteroscopy in Asymptomatic Postmenopausal Patients: indications and outcomes of 822 Cases.

Study ObjectiveThis study aims to assess the prevalence of malignancy and other endometrial pathologies in asymptomatic postmenopausal women referred for office hysteroscopy, identify main referral indications, and assess their relationship with the risk of malignancy.Secondary objectives included evaluating the association between ultrasound variables and malignancy risk and assessing procedure validity, which encompasses duration, feasibility and patient comfort during office hysteroscopy. DesignRetrospective analysis. SettingThe study was conducted at the Department of Gynecology, Division of Gynaecology and Obstetrics, University Medical Centre Ljubljana, Slovenia's largest tertiary care center. ParticipantsThe cohort consisted of 822 asymptomatic postmenopausal women referred for office hysteroscopy, excluding those with postmenopausal bleeding within the last year. InterventionsParticipants underwent office hysteroscopy with or without biopsy. Measurements and Main ResultsThe main indication for hysteroscopy was ultrasound abnormalities alone, with remaining indications including a combination of ultrasound and clinical findings. Among the cohort, 97.4% exhibited benign findings, while 2.6% were diagnosed with cancer or precancerous lesions. The analysis revealed that patients with indications based on ultrasound and clinical findings suggestive of malignancy had a higher risk of malignancy compared to those with ultrasound alone. In 387 patients with documented ultrasound variables, inhomogeneous endometrial appearance (OR: 8.2, 95% CI: 2.4-27.9, p<0.001) and significant liquid content within the uterine cavity (OR: 10.2, 95% CI: 3.6-28.9, p<0.001) exhibited strong associations with malignancy. Analysis of the procedure revealed a high feasibility rate (87.8%), with a median duration of 13,7 minutes and a median VAS pain score after the procedure of 3/10. ConclusionsThe prevalence of endometrial cancer and precancerous lesions in asymptomatic postmenopausal patients is likely low, with most intrauterine pathologies being benign. Our study demises the utility of routine endometrial surveillance for this population in the absence of specific risk factors. A holistic approach, considering individualized assessments and factors beyond endometrial thickness, is crucial in interpreting ultrasonic findings.

Clinical and diagnostic evidence of benign endometrial pathology in postmenopausal patients

The objective: to study clinical and diagnostic features of the benign endometrial conditions in the postmenopausal women.Materials and method. 64 postmenopausal patients with various severity of postmenopausal vaginal bleeding were examined. All patients underwent clinical, radiological, laboratory and histological examinations, reproductive history data and body mass index indicators were evaluated. Of them 33 persons were diagnosed with complex endometrial hyperplasia without atypia, 16 women – endometrial polyp and 15 women – hyperplasia with atypia. Routine gynecological examination including abdominal and pelvic examination following transvaginal ultrasound examination for the determination of the uterine and ovarian volume as well, as the endometrial thickness were conducted. Patients included in the study underwent computer tomography and magnetic resonance imaging as indicated. All patients with endometrial pathology underwent endometrial biopsy by standard dilatation and curettage or Pipelle-biopsy with histological examination of the obtained material. The concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), dehydroepiandrosterone sulfate (DHEA-S), estradiol, estrone, progesterone, prolactin, and testosterone were determined in blood serum.Results. It was confirmed that in 87,5% of cases endometrial hyperplasia manifested with vaginal bleeding. Endometrial thickness in examined persons ranged from 14.1±1.6 mm in patients with atypical endometrial hyperplasia to 21.3±4.8 mm in patients with complex endometrial hyperplasia without atypia. The study of blood levels of hormones found that postmenopausal patients with endometrial hyperplasia have lower FSH concentration, LH/FSH ratio, estradiol, testosterone levels, with significantly high values of prolactin, DHEA-S and estrone. During the postmenopausal period, hyperplasia was developed in 10% of cases by the presence of bleeding, and 86.2% of cases by the presence of blood spotting. Conclusions. The study suggests that high body mass index, numerous artificial abortions, high blood levels of estrone, DHEA-S and prolactin as well as increased thickness of endometrium influence the frequency of endometrial hyperplasia in postmenopausal women.

Rapid and random-start endometrial preparation before outpatient hysteroscopic polypectomy in patients of perimenopausal age

Objective This study aimed to assess the efficacy of randomly started oral dienogest/ethinylestradiol (DNG/EE) for swift endometrial preparation prior to outpatient hysteroscopic polypectomy in perimenopausal women. Method A multicenter, prospective, randomized controlled trial was conducted in university hospitals. Eighty perimenopausal women scheduled for outpatient hysteroscopic polypectomy between January 2023 and March 2024 were randomly assigned to either intervention (n = 40) or control (n = 40) groups. Exclusion criteria included concomitant endometrial pathologies, recent therapy and adnexal diseases. The intervention group received oral DNG/EE 2 mg/0.03 mg/day started on any day of the menstrual cycle for 14 days. The control group underwent polypectomy between menstrual cycle days 8 and 11 without pharmacological treatment. Results Pre-procedure (p < 0.001) and post-procedure (p < 0.001) endometrial thickness were significantly reduced in the intervention group, along with a higher incidence of hypotrophic/atrophic endometrial patterns (p < 0.001). Surgical parameters also differed significantly between groups. Conclusion DNG/EE treatment offers rapid, cost-effective endometrial preparation, enhancing surgical outcomes and patient satisfaction during outpatient polypectomy. Trial registration ClinicalTrials.gov NCT06316206.

Quantitative Assessment of Polarization and Elastic Properties of Endometrial Tissue for Precancer/Cancer Diagnostics Using Multimodal Optical Coherence Tomography

Objectives: The most important phase in the endometrial pathologies diagnostics is the histological examination of tissue biopsies obtained under visual hysteroscopic control. However, the unclear visual diagnostics characteristics of subtle focal endometrial pathologies often lead to selection errors regarding suspicious endometrial lesions and to a subsequent false pathological diagnosis/underestimation of precancer or early-stage cancer. Methods: In this study, we investigate the potential of Multimodal Optical Coherence Tomography (MM OCT) to verify suspicious endometrial lesion regions before biopsy collection. We study the polarization (by cross-polarization OCT, CP OCT) and elastic (by compression OCT-elastography, C-OCE) properties of ex vivo endometrial tissue samples in normal conditions (proliferative and secretory phases to the menstrual cycle, atrophic endometrium) with endometrial hyperplasia (non-atypical and endometrial intraepithelial neoplasia) and endometrial cancer subtypes (low-grade, high-grade, clear cell and serous). Results: To the best of our knowledge, this is the first quantitative assessment of relevant OCT parameters (depth-resolved attenuation coefficient in co-[Att(co) values] and cross-[(Att(cross) values] polarizations and Young’s elastic modulus [stiffness values]) for the selection of the most objective criteria to identify the clinically significant endometrial pathologies: endometrial intraepithelial neoplasia and endometrial cancer. The study demonstrates the possibility of detecting endometrial pathologies and establishing optimal threshold values of MM OCT criteria for the identification of endometrial cancer using CP OCT (by Att(co) values = 3.69 mm−1, Sensitivity (Se) = 86.1%, Specificity (Sp) = 92.6%; by Att(cross) values = 2.27 mm−1, Se = 86.8%, Sp = 87.0%) and C-OCE (by stiffness values = 122 kPa, Se = 93.2%, Sp = 91.1%). The study also differentiates endometrial intraepithelial neoplasia from non-atypical endometrial hyperplasia and normal endometrium using C-OCE (by stiffness values = 95 kPa, Se = 87.2%, Sp = 90.1%). Conclusions: The results are indicative of the efficacy and potential of clinical implementation of in vivo hysteroscopic-like MM OCT in the diagnosis of endometrial pathologies.

Role of Receptor for Advanced Glycation End-Products in Endometrial Cancer: A Review.

Endometrial cancer (EC) is the most common gynecological malignancy. EC is associated with metabolic disorders that may promote non-enzymatic glycation and activate the receptor for advanced glycation end-products (RAGE) signaling pathways. Thus, we assumed that RAGE and its ligands may contribute to EC. Of particular interest is the interaction between diaphanous-related formin 1 (Diaph1) and RAGE during the progression of human cancers. Diaph1 is engaged in the proper organization of actin cytoskeletal dynamics, which is crucial in cancer invasion, metastasis, angiogenesis, and axonogenesis. However, the detailed molecular role of RAGE in EC remains uncertain. In this review, we discuss epigenetic factors that may play a key role in the RAGE-dependent endometrial pathology. We propose that DNA methylation may regulate the activity of the RAGE pathway in the uterus. The accumulation of negative external factors, such as hyperglycemia, inflammation, and oxidative stress, may interfere with the DNA methylation process. Therefore, further research should take into account the role of epigenetic mechanisms in EC progression.

Assessment of magnetic resonance imaging findings in ovarian granulosa cell tumors along with clinical prognostic factors.

To determine the role of preoperative MRI in the diagnosis and treatment of patients with granulosa cell tumors (GCTs) of the ovary. Twenty-four patients who were operated on between 2018 and 2022 and who were pathologically diagnosed with GHT and met the inclusion criteria were retrospectively examined. The findings were compared with the patients' demographic data, symptoms, surgical findings (laterality, stage, lymph node involvement, endometrial pathology, tumor size), and CA-125 levels. The final cohort included 24 patients with a mean age of 54.71 ± 16.52. All the patients had the pathological diagnosis of adult type GCT. In the morphological evaluation, the most common finding was a solid-cystic mixed type (14 patients, 58.3%), while intratumoral hemorrhage signal was observed in 10 patients (41.7%). In the majority of cases (91.7%), the mass showed regular contours. The honeycomb/Swiss cheese sign was detected in 54.2% of the cases. When the T1 and T2 signal of the solid component of the mass were examined relative to the myometrium, the majority of GCTs appeared isointense on both sequences (83.3% and 62.5%, respectively). The mean ADC value of the solid component obtained from diffusion-weighted imaging was 0.78 ± 0.15 × 10-3. Pelvic fluid was observed in 41.7% of the cases. The average endometrial thickness was 9.74 ± 6.43 mm. Thickened endometrium more than 9 mm was observed in 9 out of the remaining 21 patients (42.9%). Understanding the key imaging features for GCTs plays an essential role in the diagnosis and guiding the treatment effectively.

The Role of Transabdominal, Transvaginal and Doppler Ultrasound in the Assessment of Endometrial Pathology

The Role of Transabdominal, Transvaginal and Doppler Ultrasound in the Assessment of Endometrial Pathology

Endometrial pathology in abnormal uterine bleeding (AUB) &amp; role of different techniques of endometrial sampling in evaluation of AUB: A cross-sectional study

: Abnormal uterine bleeding (AUB), traditionally defined as uterine bleeding that is abnormal in volume, regularity, and/or timing. It affects 14–25% of women of reproductive age. AUB has a major impact on quality of life of women. Aetiology of AUB may be structural and functional and varies with the age group. Histology of endometrium remains the mainstay in diagnosis of pathology and management of AUB. Various techniques such as endometrial sampling by pipelle, dilatation and curettage (D &amp; C) and hysteroscopy are used to obtain endometrium for histological diagnosis. A single center prospective cross-sectional study at teaching institute was carried out to include 150 females from 18-70 years presenting with abnormal uterine bleeding. Objective of study was to analyse prevalence of various endometrial pathologies in AUB, symptomatology as well as age wise distribution of various endometrial pathologies in AUB and compare feasibility of different techniques of collecting endometrial sample. Endometrial sample was obtained either by OPD Pipelle brush or Surgical Dilatation and Curettage method or hysteroscopy. Statistical Analysis was done by using Epi-Info-7 software.Commonest age group presenting with AUB was 41-50 (40%) years, 64% were para 3 or more, Proliferative and secretary changes, 53(35.3%) and 24(16%) cases while endometrial hyperplasia without atypia 18 (12%) cases and with atypia 7 cases (4.7%), endometrial carcinoma 5 cases (3.3%) while endometrial polyp was seen in 8 cases (5.3%). Heavy menstrual bleeding with normal duration was the commonest presentation in 64%. Endometrial sampling technical failure was seen in 2 cases with pipelle biopsy while in 7 cases inadequate sample was obtained with pipelle and dilatation and curettage. Uterine perforation was seen in one case which was managed conservatively.Technical failure and inadequate sample needs to be addressed considering background high risk factors, TVS imaging, repeat sample and use of hysteroscopy may be offered case to case basis.

COVID-19 vaccination and postmenopausal bleeding: a retrospective cohort study

Objective COVID-19 vaccination has been related to menstrual irregularities; however, the effect on postmenopausal women is unknown. The aim of this study was to analyze the prevalence of postmenopausal bleeding (PMB) after COVID-19 vaccination. Methods A retrospective study was conducted in the Department of Gynecology in Hospital del Mar. Consecutive postmenopausal women with data available and endometrial biopsy were included between February 2021 and January 2022. Patients were stratified between COVID-19 vaccinated and unvaccinated groups. PMB after 30 days from last vaccine dose was considered unrelated to vaccine. Endometrial pathology diagnoses were stratified into benign or malignant. Univariable and multivariable of regression analysis on variables potentially associated with PMB was performed. Results A total of 381 patients were included, 91 in the vaccinated group and 290 in the unvaccinated group. Prevalence of PMB in the vaccinated group was 75.8% compared to 59.0% in the unvaccinated group (p < 0.005). No increase in endometrial malignant pathology was observed among the vaccinated group (p = 0.189). Multivariable analysis that correlates factors associated with PMB suggests COVID-19 vaccine and malignant endometrial biopsy as independent risk variables. Conclusions A higher prevalence of PMB was associated with COVID-19 vaccine. Endometrial histological results showed no association with COVID-19 vaccination, but endometrial biopsy should be performed for PMB.

Generation and Characterization of Rat Uterus Organoids from Rat Endometrial Epithelial Stem Cells.

Endometrial organoids offer valuable insights into the development and pathophysiology of endometrial diseases and serve as platforms for drug testing. While human and mouse endometrial organoids have been developed, research on rat endometrial organoids remains limited. Given that rats can better simulate certain endometrial pathologies, such as intrauterine adhesions, this study aimed to establish rat endometrial organoids. We present a detailed protocol for the isolation and culture of rat endometrial epithelial stem cells (reESCs) and the generation of rat endometrial organoids. Using a refined reESCs expansion medium, we successfully isolated and stably expanded reESCs, demonstrating their long-term culture potential. The reESC-generated organoids exhibited typical structural and functional characteristics of the endometrium, including hormone responsiveness. Our results showed that rat endometrial organoids could be cultured over a long term with stable proliferation, maintaining the glandular structure, cell polarity, and functional characteristics of the endometrial epithelium. This novel rat-derived endometrial organoid model provides a valuable platform for studying endometrial diseases and testing therapeutic interventions, with potential applications across various mammalian species.

Hysteroscopic Endometrial Fundal Incision versus Hysteroscopy Only in Oocyte Recipients: A Randomized Controlled Trial Assessing The Reproductive Outcomes.

Endometrial scratching (ES) remains controversial regarding its potential effectiveness in improving pregnancy rates. The objective of the present study was to assess the impact of endometrial fundal incision (EFI) during hysteroscopy on reproductive outcomes in a population of oocyte recipients. A randomized controlled trial was conducted between 2020 and 2023 at the Third Department of Obstetrics and Gynecology, School of Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki and "Assisting Nature Centre of Reproduction and Genetics". The study population consisted of women who underwent hysteroscopy randomly assigned in a 1:1 ratio to either EFI (one to three months before embryotransfer with donor oocytes) or no intervention throughout office hysteroscopy. Clinical pregnancy and live birth rates were the primary outcomes. After the exclusion of patients with intraoperative diagnosed endometrial pathology, a total of 124 women underwent randomization. The pregnancy test was positive in 79% (n=49/62) of the women in the EFI compared to 59.7% (n=37/62) in the hysteroscopy-only group (P=0.019), while the live birth rates did not differ between the two groups (58.1%, n=36/62 vs. 51.6%, n=32/62, P=0.470). EFI during hysteroscopy seems to improve pregnancy rates in oocyte recipients without intrauterine pathology, while live birth rates are not affected by the EFI. These results should be interpreted with caution before the implementation of EFI in the routine in vitro fertilization (IVF) practice (registration number: NCT04580056).

P-792 Human endometrial organoids embedded in hybrid endometrial-derived hydrogels as a biocompatible treatment for endometrial pathologies in preclinical models

Abstract Study question Are human endometrial organoids (hEOs) labelled with Molday-ION Rhodamine B (MIRB) within hybrid endometrial-derived hydrogels a biocompatible approach to treat endometrial pathologies in immunocompetent mice? Summary answer The direct intrauterine injection of hEOs labelled with MIRB within hybrid endometrial-derived hydrogels demonstrated xenogeneic biocompatibility in preclinical models over a 2-week period. What is known already The lack of effective treatments for endometrial pathologies emphasizes the urgent need for novel bioengineering approaches. In this context, hEOs are presented as promising candidates for tissue regeneration, since they offer a native and partially undifferentiated cellular source with high proliferative capacity, which is fostered after its culture within hybrid endometrial-derived hydrogels (Gómez-Álvarez et al. 2023). Their concurrent injection aims to enhance proliferation and differentiation through its rich biochemical composition. No prior study has successfully labelled and injected hEOs in immunocompetent models, which is crucial to discern whether the therapeutic effect operates at paracrine level or involves direct tissue regeneration. Study design, size, duration This study was approved by an ethics committee. Decellularized porcine endometrium was used to create endometrial-derived hydrogels as previously described and then mixed with PuraMatrix to obtain the hybrid endometrial-derived hydrogel. After 7 days of culture, hEOs (passages 6-7) were labelled with MIRB for final tracking. Finally, hEOs were mixed with the hybrid hydrogel and injected into the uterine horns in immunocompetent mice (n = 3). After two weeks, biocompatibility was evaluated through blood and macroscopical analyses. Participants/materials, setting, methods After 7 days, hEOs were collected and labelled with MIRB (50 µg/ml) for their final tracking in the murine endometrium. Endometrial-derived hydrogel (8 mg/ml) was mixed with PuraMatrix 1% (50:50 ratio) to generate the hybrid hydrogel. After its characterization and in vitro validation (Gómez-Álvarez et al. 2023), it was used as the bioactive carrier. Each animal received a single intrauterine injection (50 µl, ∼100.000 cells). Blood, spleen, and uterus were collected after 2-week period. Main results and the role of chance Hybrid endometrial-derived hydrogels demonstrated biocompatibility and in vivo stability in mice over a 2-week period (Gómez-Álvarez et al. 2023). In this study, we aim to use this hybrid hydrogel with hEOs as a biocompatible treatment in preclinical models. After 7 days of culture, hEOs were successfully labelled with MIRB (∼85% efficacy), which was confirmed under fluorescence microscopy. The single intrauterine injection (50 µl, ∼100.000 cells) did not cause apparent adverse immune reactions in mice (n = 3) during the 2-week period. All mice exhibited no signs of distress and their behavior remained within normal parameters. Furthermore, the macroscopic examination of uterine horns revealed normal appearance, characterized by the absence of anomalies and tumors. At day 14 post-injection, haematocrit analyses showed normal white blood cell count: 0.89 ± 0.53 · 109/L neutrophils; 7.82 ± 2.23 · 109/L lymphocytes; 0.21 ± 0.13 · 109/L monocytes; 0.04 ± 0.04 · 109/L eosinophils; 0.00 ± 0.00 · 109/L basophils. In addition, no significant changes in red blood cells (9.31 ± 0.54 · 1012/L) and platelets (1063.00 ± 318.58 · 109/L) were observed. Histological analyses will be perform to assess the presence of hEOs labelled with MIRB in the murine endometrium and dilucidate their mechanism of action. Limitations, reasons for caution This study has been carried out in healthy C57BL/6 mice. To confirm the scope of this technique, the next step is to explore the regenerative potential of hEOs labelled with MIRB within hybrid endometrial-derived hydrogels in murine models with endometrial damage. Wider implications of the findings These findings demonstrate the biocompatibility of hEOs labelled with MIRB within hybrid endometrial-derived hydrogels in immunocompetent C57BL/6 mice. This bioengineering approach holds the potential to improve the treatment landscape for specific endometrial pathologies. Trial registration number Not applicable