Expression Of Receptivity Markers Research Articles

Study of LIF, NR3C1, BCL6, miR-494, and miR-30d in Endometrial Epithelial Cells of Repeated Implantation Failure and Healthy Women Treated with Calcitriol and Letrozole in a Fibrin Scaffold

Background: Determining the exact time of window of implantation (WOI) in individuals with repeated implantation failure (RIF) is necessary for successful implantation. The present study aimed to compare the effects of letrozole and/or calcitriol on the expression of receptivity markers in endometrial epithelial cells (EECs) of RIF patients and healthy fertile women. Methods: Endometrial tissue was obtained from RIF patients (n=5) and healthy women (n=5) in the proliferation phase. EECs were isolated and cultured in a fibrin scaffold in 10 groups. The cells were treated with letrozole and/or calcitriol in three types of treatment. To check the proliferation and migration of cells in the scaffold, immunofluorescent and H&E staining was used, and gene expression was checked using real-time polymerase chain reaction (PCR). Results: This study showed that the expression levels of pro-implantation genes (NR3C1, LIF, and miR-30d) were higher in healthy women than in RIF patients, and the expression levels of anti-implantation genes (BCL6 and miR-494) were lower in healthy women than in RIF patients. Calcitriol and letrozole increased the expression level of pro-implantation genes (NR3C1, LIF, and miR-30d) in RIF patients. Furthermore, they reduced the expression level of anti-implantation genes (BCL6 and miR-494) in RIF patients. Conclusion: Our findings suggest that treatment with letrozole is more effective than co-treatment with letrozole and calcitriol or alone treatment with calcitriol on receptivity markers in RIF patients.

Morphological features of endometrial pinopodia formation during the luteal phase in women with previous unsuccessful implantation

Pinopodia of the receptive endometrium is one of the prerequisites for successful embryo implantation.Study objective: to analyze the morphological changes of the endometrium in infertile women, in particular to investigate the apical surface of epithelial cells and pinopodia formation. Materials and methods. 12 patients with a history of previous implantation failure were examined. Patients underwent hysteroscopy with biopsy under intravenous anesthesia on 20–24th day of the menstrual cycle. Morphological structure of pinopodia on the surface of the endometrial epithelium was evaluated by scanning electron microscopy.Results. Apical surface of epitheliocytes was damaged in 16.7% studied biopsies. Morphological picture and maturity of the endometrial glands did not correspond to the actual day of the menstrual cycle in 41.7% biopsies. Degree of glands maturity in these patients was low, so there was a delay in the menstrual cycle. Ultrastructural examination showed clear changes in the structure of secretory and ciliary cells in 33.3%. On the apical surface of the secretory cells there was no or small number of microvilli; cilia of eyelashes were shortened or sometimes fused together. Pinopodia as a one of the most important components of implantation was detected in only 50% of patients. Only in 33.3% of participants visualized fully developed, mature pinopodia. Conclusions. Analysis of own data showed that in the endometrium of women with previous unsuccessful implantation there are certain morphological changes, which are physiologically manifested by a delay in the luteinizing phase of the menstrual cycle and, possibly, as a result mismatch of endometrial structure and embryo maturity. Time of pinopodia onset in participants varied from 20 to 24 days of the menstrual cycle. It is necessary to improve the design and conduct further research to study the association between the expression of receptivity markers and ultrastructural structure.

Platelet-Rich Plasma Improves Pregnancy Rate and Repairs Endometrial Injury in Patients with Repeated Implantation Failure

Objective: The effects investigated in this study for the therapy with autologous platelet-rich plasma (PRP) on the thin endometrium in a rat model and patients with repeated implantation failure (RIF). Methods: PRP were immediately injected into uterine cavity after the establishing a model of thin endometrial injury by injection with 95% ethanol into uterus of SD rat. We have used H&E staining to explore the endometrial morphological alteration. The immunohistochemistry, Western blots, and quantitative RT-PCR were used to determine the endometrial receptivity. RL95-2 cells were incubated with RPR at the different concentrations to detect the effect of PRP on the endometrial epithelial cell proliferation. Patients (n = 51) were divided into the control and PRP treatment groups. Patients in the PRP treatment group received PRP by intrauterine perfusion. Results: Endometrial morphology was significantly improved after PRP intrauterine infusion thrice-administered SD rats. PRP increased the thickness of thin endometrium in rats and up-regulate the expression of receptivity markers, including vimentin, vascular endothelial growth factor (VEGF) and integrin β3. In the control (n = 25) and PRP treatment (n = 26) groups, no significant differences were observed in rates of clinical pregnancy (50.0% vs. 44.0%, p = 0.67), implantation (40.5% vs. 26.0%, p = 0.15) and miscarriage (30.8% vs. 18.2%, p = 0.48). Conclusions: The transplantation of PRP repairs the endometrial injury by suppressing receptivity, enhancing endometrial cell proliferation and vascular remodeling. PRP improves the pregnancy rate in RIF patients.

Effects of noncavity-distorting fibroids on endometrial gene expression and function.

Uterine fibroids are a common finding in infertility patients. Impaired implantation and decidualization have been proposed to contribute to compromised fertility. Data are limited on the endometrial transcriptome from subjects with uterine fibroids, as well as endometrial receptivity and decidualization potential of endometrial stromal fibroblasts (eSF) from women with fibroids. Our objective was to investigate the endometrial transcriptome of women with noncavity-distorting intramural fibroids and compare them to control subjects with no uterine pathology throughout the menstrual cycle. We also evaluated endometrial receptivity gene expression and basic endometrial functions such as decidualization, proliferation, and apoptosis in women with fibroid uterus. Results showed that large numbers of transcripts were significantly dysregulated throughout the menstrual cycle in fibroid subjects compared to controls. However, there were essentially no differences in the expression of receptivity markers at the tissue level, as well as decidualization markers in tissue and eSF in subjects with fibroids compared to controls. However, eSF from women with a fibroid uterus exhibited decreased proliferation potential and increased apoptosis upon decidualization. These data indicate preserved implantation and decidualization potential despite observed gene expression changes in endometrium from women with noncavity-distorting fibroids compared to controls. How this phenomenon and altered proliferation/apoptosis may contribute to impairment of endometrial function in subfertile patients warrants further investigation.

Endometrial receptivity markers in infertile women stimulated with letrozole compared with clomiphene citrate and natural cycles

Though various advantages associated with the use of letrozole as an ovulation inducing drug are documented, there is inadequate information regarding its effect on endometrial receptivity. Expression of endometrial receptivity markers including αvβ3 integrin, L-selectin, LIF, and pinopods during the implantation window in infertile women stimulated with letrozole or clomiphene citrate (CC), and spontaneous cycles is investigated. A total of 36 women were included in the study out of which 16 women were diagnosed with primary unexplained infertility and the other 20 women with severe oligoasthenozoospermic/azoospermic male partners. Both groups were sub-divided into three groups; women stimulated with letrozole, or with CC, and the third group was allowed to ovulate spontaneously (natural cycles). Women having natural cycles and not given any drug were considered as controls. Once ovulation was confirmed by ultrasonography, endometrial samples were collected on the seventh day post-ovulation and analyzed. In women with unexplained infertility treated with letrozole and CC, epithelial and stromal expression of αvβ3 integrin, L-selectin, leukemia inhibitory factor (LIF), and pinopod formation was found to be significantly higher as compared to controls. Expression of these receptivity markers was found to be comparable amongst the letrozole, CC, and control groups in women with severe oligoasthenozoospermic/azoospermic male partners. Cell cycle analysis showed similar cell cycle phase fractions on comparing the CC and the letrozole groups. Stimulation with letrozole and CC appears to enhance endometrial receptivity in women with unexplained infertility. However, letrozole and CC did not have any significant effect on the endometrial receptivity markers of women with severe oligoasthenozoospermic/azoospermic male partners.

HOXA-11 mediated dysregulation of matrix remodeling during implantation window in women with endometriosis

To investigate the Homeobox genes HOXA-10 and HOXA-11 mediated endometrial molecular defects during implantation window in endometriosis-associated infertility cases. Endometrial biopsies were obtained during implantation window from 31 infertile women with endometriosis (age < 35 years) and 26 age and BMI-matched infertile women without endometriosis were included in the study for comparison purposes. Endometrial expression of HOXA-10 and HOXA-11 genes, MMP-2, -9, α(v)β(3) integrin, leukemia inhibitory factor and surface characteristics including average roughness and topology were assessed. A significantly lower expression of HOXA-10 and HOXA-11 were observed in endometriotic women compared to non-endometriotic controls. Further, a significantly higher endometrial expression of MMP-2 and -9 were observed in women with endometriosis when compared with controls. Interestingly, endometrial surface were observed to be grossly affected in terms of average roughness and topology in women with endometriosis compared to controls. The findings suggest that aberrant expression of HOXA-10 and -11 genes adversely affects endometrial remodelling and expression of receptivity markers.

Effects of “Bu Shen Huo Xue Decoction” on the Endometrial Morphology and Expression of Leukaemia Inhibitory Factor in the Rat Uterus during the Oestrous Cycle

The purpose of this study was to explore the positive effects of Bu Shen Huo Xue Decoction (BSHXF) on assisted reproduction. The study aimed to evaluate whether BSHXD could improve endometrial morphology and increase the expression of LIF in a gonadotrophin-releasing hormone agonists (GnRHa) long protocol-induced rat model during metestrus, diestrus, proestrus, and oestrus. The BSHXD group presented significantly increased endometrium thickness and decreased MVD compared with the GnRHa long protocol group. In addition, the expression of LIF was significantly higher in the BSHXD group. There were no significant differences between the control group and the BSHXD group in terms of MVD and LIF expression. These results suggested that BSHXD can improve the endometrium development, reduce the abnormal angiogenesis, and increase the expression of receptivity markers in a GnRHa long protocol-induced rat model during the oestrous cycle, which might result in an endometrial environment better suited for female reproduction.

The expression of receptivity markers in the fallopian tube epithelium

Pinopodes represent the morphological and integrins, the biomolecular markers of endometrial receptivity. We studied using scanning electron microscopy, the expression of pinopodes on tubal samples and their corresponding endometria, from 21 women of reproductive age (7 from proliferative phase, 7 from day LH +5 and 7 from day LH +7). In addition, we examined the immunohistochemical staining of integrins alpha v beta 3, alpha v beta 5 and their ligands, fibronectin (FN) and osteopontin (OPN) in the same tubal epithelium samples. Pinopodes were detected on the tubal epithelium exclusively during day LH +7, coincident with their formation in the endometrium and synchronous to alpha v beta 3 sharp increase in the oviduct epithelium, suggesting a regulation similar to the endometrium. In contrast, alpha v beta 5, FN and OPN remained unchanged during the cycle. These results show for the first time the formation of pinopodes in the tubal epithelium at the time of endometrial receptivity and correlate it with the upregulation of the intact dimmer alpha v beta 3 in the tubes.

Modulation of Endometrial HOXA10 and Other Markers of Endometrial Receptivity by Human Chorionic Gonadotropin (hCG) as a Function of Uterine Age

Objective: In vitro evaluation of endometrial receptivity (ER) remains elusive and is limited by cell culture which disrupts physiologic interactions between stroma, glands, and epithelium. We have previously confirmed the utility of the endometrial explant culture system as a model for studying ER. Using this model, we sought to characterize the endometrial response to hCG, as influenced by uterine age, using receptivity markers, including HOXA10, vascular endothelial growth factor (VEGF), and glycodelin. Materials and Methods: Endometrial biopsy was performed in 9 prospective recipients of egg donation. Subjects (42±8 years) received estradiol (E2) and progesterone (P4) with a biopsy performed on day 7 of P4. Samples were cut into 1mm3 pieces and cultured in DMEM/F-12 with E2 and P4, without (control) or with hCG (10, 100, 1000ng/mL), on Millicell-CM inserts for 24 hours. Explant viability was assessed using immunohistochemistry (IHC). Semi-quantitative PCR was performed using an internal control gene to evaluate relative messenger RNA (mRNA) expression of HOXA10, VEGF, and glycodelin. Results: Explant viability was confirmed on IHC by histology and proliferation staining. A non-significant, dose-dependent increase was observed in VEGF (p= .24), HOXA10 (p = 0.07), and glycodelin (p = 0.11) in response to increasing hCG. HOXA10 trended towards a statistically significant increase between hCG of 10 and 100ng/mL (1.29±1.45 vs 2.21±2.01; p<.08). Glycodelin demonstrated a statistically significant increase between control and 10ng/mL hCG (1.00 vs 1.66±0.57; p<.05). Age was inversely associated with HOXA10 (-0.09±0.03 mRNA expression/year of age, p<.01) and positively associated with VEGF (0.04±0.02 mRNA expression/year of age, p<.05). Conclusions: 1) The endometrial explant culture system is a promising model for the study of ER, as it maintains interactions between stroma, glands, and epithelium. 2) HOXA10, VEGF, and glycodelin all demonstrated a dose-response increase in mRNA expression in response to hCG, though statistical significance was not achieved. 3) Increasing expression of ER markers in response to hCG supports the role of hCG as a candidate protein for blastocyst–endometrial communication. 4) Statistically significant associations between age and expression of receptivity markers provides the first evidence that, among recipients of egg donation, uterine age may play a role in ER on a molecular level.