Skip to main content
Download PDF

Changes in cell proliferation, but not in vascularisation are characteristic for human endometrium in different reproductive failures - a pilot study

Reproductive Biology and Endocrinology volume 8, Article number: 67 (2010) Cite this article

Abstract

Background

Reproductive failure, determined as recurrent spontaneous abortions (RSA) or recurrent implantation failure (RIF) in women is not well understood. Several factors, including embryo quality, and cellular and molecular changes in endometrium may contribute to the insufficient feto-maternal interaction resulting in reproductive failure. Prior clinical studies suggest an inadequate endometrial growth and development of the endometrium, leading to a lesser endometrial thickness.

Methods

We therefore aimed to determine the cellular proliferation using Ki67, and the expression of markers of vascularisation, such as factor VIII (a marker of endothelial cells) and smooth muscle cell actin (SMCA; a marker of pericytes and smooth muscle cells) in endometrium of healthy women and women with RSA or RIF. LH-dated mid-secretory endometrial biopsies of seven healthy women and twenty women with reproductive failure were examined via immunohistochemistry followed by image analysis.

Results

Cellular proliferation but not expression of factor VIII or SMCA was decreased (P < 0.0004) in endometrium of women with RSA and RIF compared to healthy controls. Conclusion: Our data indicate that reproductive failure is due to insufficient cell proliferation/tissue growth rather than inadequate vascularisation in the endometrium.

Background

Reproductive failure comprises idiopathic recurrent spontaneous abortions (RSA) as well as recurrent implantation failure (RIF) [1]. One of the current RIF definition is a failure to conceive after three transfers of one or more good quality embryos; however, this definition may vary depending on the center [2]. On the other hand, idiopathic RSA is defined as three or more consecutive pregnancy losses within the first 20 weeks of gestation after exclusion of known contributing factors including uterine malformations, bleeding abnormalities, hormone disequilibrium, parental chromosomal defects, infections and others [3].

Adequate implantation is a limiting factor in human reproduction. Despite major improvement in assisted reproduction techniques (ART) the clinical pregnancy rate per embryo transfer in fresh ART cycles is 31%, and up to 41% in oocyte donation programs [4]. A number of etiologies, including decreased embryonic quality, endometrial receptivity, feto-maternal communication, endocrine, genetic and other factors have been suggested as causes for reproductive failure [5–7]. Even though intrauterine endometrium is not essential for embryo implantation, since ectopic pregnancies occur [8], several studies indicated an altered endometrial function in women with reproductive failure [9, 10]. Changes at the cellular and molecular level in endometrium from women with reproductive failure have been reported [8, 11, 12]; suggesting an impact of specific genes on the success of implantation. However, these studies determine mostly gene expression profiles and only partially examine the transcribed products.

Although inadequate regulation of endometrial growth has been recognized as a possible factor in reproductive failures [11, 13–15], limited data is available concerning endometrial cell proliferation and its regulation. Furthermore, inconsistent results were obtained from studies of the role of endometrial vascularisation, angiogenesis, blood flow and/or endometrial thickness in women with implantation failure; while some studies demonstrate differences between normal and reproductive failures subjects others do not [16–19].

We hypothesized that reproductive failures are due to altered uterine growth caused by changes in cell proliferation and vascularisation/angiogenesis in the endometrium. Therefore, the aim of this study was to examine cellular proliferation and expression of markers of vascularisation (factor VIII and smooth muscle cell actin [SMCA]) at the protein level in endometrial biopsies from the mid-secretory phase of healthy women and women with reproductive failure.

Methods

In this observational, non-therapeutic pilot study, endometrial biopsies were taken after informed consent according to the Ethical Committee of the University of Heidelberg, Germany. A total of 27 women, at age 25 to 42 years, who were not on any hormonal treatments were included in this study. All women exhibited regular 28 ± 1 day cycles. Pipelle biopsies were taken in the mid-secretory phase on day 8-9 after the LH surge (LH +8/+9) from 11 women with RIF, 9 women with RSA, and 7 healthy woman. RIF was defined by failure to detect a positive serum hCG after three transfers of two or three good quality embryos created through in vitro fertilization. RSA was defined as the appearance of three or more spontaneous abortions of unknown reasons during the first trimester. None of the women with RIF or RSA had ever had a successful term pregnancy. Control women delivered healthy babies at term, with one woman having had a preterm delivery due to premature rupture of membranes. None of the women included in the study had known contributing factors for pregnancy failure, including thrombophilia (Factor V Leiden mutation, prothrombin mutation, antithrombin III, as well as protein C or S deficiency), antiphospholipid syndrome (anticardiolipin antibodies, lupus anticoagulans, ANAs), uterine anomalies detected by ambulatory hysteroscopy, polycystic ovarian syndrome, hormone abnormalities (thyroid, prolactin etc.) and other endocrinopathies (e.g., diabetes). On the day of biopsy blood samples were collected for determination of estradiol-17β (E2) and progesterone concentration in serum using a competitive immunoassay by the University of Heidelberg core facility. The LH surge was determined using a commercially available urine LH kit (Clear blue, WICK PHARMA/Procter & Gamble GMBH, Schwalbach am Taunus, Germany). Furthermore, histology of the biopsies typical for the secretory phase was confirmed by two individual researchers according to the modified Noyes' criteria [20].

Tissue preparation

Immediately after the biopsies were performed, tissues were immersed in OCT, frozen in liquid nitrogen and stored at -80°C. Tissues were then sectioned at 8 μm, mounted onto SuperFrost Plus slides (Menzel GmbH & Co, Braunschweig, Germany), fixed with 100% acetone at 4°C for 10 min, and stored at -70°C before immunohistological procedure.

Immunohistochemistry

Cell proliferation was determined based on immunolocalization of Ki67 protein (a marker of proliferating cells) and vascularisation was determined by immunolocalization of factor VIII (a marker of endothelial cells) and SMCA (a marker of pericytes and smooth muscle cells) as previously described [21, 22]. Briefly, sections of cryopreserved endometrial tissues were rinsed several times in PBS containing Triton-X100 and were then treated for 20 min with blocking buffer [PBS containing normal goat serum (1-2%, v/v)] followed by treatment with primary antibody against Ki67 (1:100; mouse monoclonal; Vector Laboratories, Burlingame, CA, USA), factor VIII (1:100; rabbit polyclonal; Sigma, St. Louis, MO, USA) or SMCA (1:150; mouse monoclonal; Oncogene Research Products; San Diego, CA, USA) overnight at 4°C. Primary antibody was detected using a biotin-labeled secondary antibody (anti-mouse antibody for Ki67 and SMCA, and anti-rabbit antibody for factor VIII; Vector Laboratories) and the ABC kit (Vector Laboratories). For color development, Vector SG substrate kit (Vector Laboratories) was used. Sections stained for the presence of Ki67 were counterstained with fast red (Sigma) to visualize cell nuclei. Control sections were incubated with normal mouse IgG (4 μg/mL) or rabbit IgG (diluted 1:100) in place of primary antibody.

Image analysis

Image analysis was performed as described in detail previously [22, 23]. Endometrial images of randomly chosen areas (5-10 per uterine section/subject; 0.025 mm2 per field) stained for Ki67, factor VIII or SMCA were taken at 400 × magnification, using the Eclipse E600 Nikon microscope and digital camera. Labeling index (LI; proportion of proliferating Ki67-postive cells out of the total cells per area), vascularity based on relative expression of factor VIII (occupied by endothelial cells) and SMCA (located in smooth muscle cells and pericytes) were determined by using computerized image analysis (Image-Pro Plus, version 5.0; Media Cybernatics, Houston, TX, USA). For factor VIII and SMCA, data are expressed as the total area that exhibited positive staining within tissue area.

Statistical Analysis

Data were analyzed using the general linear model (GLM) procedures of SAS (SAS Inst. Inc. Cary, NC). When the F-test was significant (P < 0.05), differences among means were evaluated by using the least square means procedure [24]. Data are expressed as mean ± SEM.

Results

The mean age of the women in the control group (30.3 ± 1.9 years) was similar to the women with RSA (34.6 ± 1.2 years) and the women with RIF (34.3 ± 0.8 years). E2 and progesterone serum concentration were similar in control, RSA and RIF (E2: 146 ± 21, 158 ± 25 and 128 ± 11 pg/ml; progesterone: 14.4 ± 4.3, 11.0 ± 1.7 and 10.2 ± 0.8 ng/ml), respectively.

Ki67, factor VIII and SMCA were detected in uterine tissue sections in all groups (Fig. 1 and 2). Ki67 was detected in cells of uterine glands, stromal tissue and blood vessels (Fig. 1A-C), while factor VIII (Fig. 2A) and SMCA (Fig. 2B) were localized to blood vessels. Factor VIII was detected in small and larger blood vessels (Fig. 2A), and SMCA was detected mostly in the larger blood vessels, but also in some small blood vessels (Fig. 2B).

Figure 1
figure 1

Immunohistochemical staining of Ki67. Representative micrographs of positive staining of Ki67 in human endometrium obtained from healthy women (A), and women with RSA (B) or RIF (C). Dark staining indicates proliferating cells, and pink staining indicates cell nuclei of non-proliferating cells. Note more proliferating cells in A than in B or C. Proliferating cells in endometrial glands are marked with small arrows in A and B, and in stromal tissue with large arrows in A, B and C. Insert in C demonstrated proliferating cells in a blood vessel. Endometrial glands are marked with stars (*). Control staining did not show any positive staining for Ki67 (D). Bar = 50 μm.

Full size image
Figure 2
figure 2

Immunohistochemical staining of factor VIII and SMCA. Representative micrographs of positive staining (dark color) of factor VIII (A) and smooth muscle cell actin (B) in human endometrium obtained from healthy women. Note positive staining in small (small arrows) and larger (large arrows) blood vessels. Endometrial glands are marked with stars (*). Control staining did not show any positive staining for factor VIII and SMCA (insert in B). Bar = 50 μm.

Full size image

The labeling index was greater (P < 0.0004) in endometrium of the control group compared to the endometrium of women with RSA or RIF, which were both similar (Fig. 3). The cell proliferation index of the different groups was not significantly correlated to the estrogen serum concentration (P = 0.369). The expression of factor VIII or SMCA in endometrium was similar in all three groups (Fig. 3).

Figure 3
figure 3

Relative Expression of assessed factors in the control and study groups. Labeling index (LI), expression of factor VIII and SMCA in endometrium of the control (black bars), RSA (open bars) and RIF (grey bars) groups. a,bP < 0.0004 for LI.

Full size image

Discussion

The present data demonstrates that in two pathological groups, RSA and RIF cellular proliferation is less than in a control group. However, vascularisation of endometrium in these pathological groups was similar to the control group.

Although it is widely accepted that the quality of the embryo is the essential factor for successful pregnancy outcome, several studies have shown the importance of endometrial function in successful pregnancy outcome [5, 14, 25]. Numerous studies have focused on causes of implantation failures by examining embryo effects or endometrium at cellular and molecular levels [8, 11, 12, 26, 27]. The function of the endometrium is to allow implantation on one hand, but make sure that invasion of the uterine tissue is limited, as otherwise inadequate placentation occurs, including placenta accreta, increta, percreta [28]. Furthermore, endometrial cell proliferation and differentiation has to be tightly controlled [14], as progression to endometrial cancer can be seen when uninhibited proliferation takes place [29]. In the present study, inadequate endometrial growth marked by reduced cell proliferation in endometrium could contribute to reproductive failure. Although very few studies investigated endometrial cell proliferation and its regulatory mechanisms, decreased proliferation is found in endometrium at the time of menopausal transition in women [30], while in endometriosis, cellular proliferation in the human endometrium was enhanced around the time of implantation and during late secretory phase [31]. Furthermore, Mertens et al have described ongoing tissue proliferation of endometrial stromal cells via Ki67 expression in the secretory phase despite rising progesterone levels [32]. In agreement with our results, Lee and colleges [11] have indicated that defective cell growth, regulated by the differentially expressed genes controlling the cell cycle, is likely to contribute to implantation failure.

Numerous growth factors of uterine tissues including fibroblast growth factors, epidermal growth factors, vascular endothelial growth factor and others are involved in the control of endometrial growth and function [33, 34]. In addition, estrogens and progesterone are also important regulators of endometrial function [15, 35]. However, in our study as well as in other studies, concentration of E2 and progesterone were similar in normal and pathological conditions [36]. This suggests that inadequate expression of growth factor(s), rather than steroid hormones contributed to decrease the cell proliferation in reproductive failure cases. Nevertheless the effect of steroid hormones cannot be excluded, since a difference in distribution of nuclear and cytoplasmic progesterone receptors was found in women with early pregnancy loss compared to women with proven fertility [36]. In addition, the progesterone receptor is thought to play a role in the implantation defect of women with RIF [37]. Last but not least a reduction of estrogen receptor alpha at the time of implantation is described in women with RIF [10]. Therefore, further investigation is needed to evaluate the potential impact of the changes in hormone receptors on endometrial cell proliferation in women with reproductive failure.

Since cellular proliferation is reflected by tissue growth and development, endometrial thickness and/or volume is likely to depend on the rate of cell proliferation at specific reproductive stages. In fact, several studies have demonstrated a positive association between pregnancy rate and endometrial volume and/or thickness [38, 39]. Thus, these observations are in agreement with our data showing reduced cell proliferation in reproductive failures. On the other hand, several studies did not observe any association between endometrial volume or endometrial thickness and pregnancy outcome [16, 40]. These discrepancies are likely due to different timing and treatments applied before measurement of endometrial volume or thickness. Therefore, more detailed and uniformed studies should be performed to determine the role of endometrial cell proliferation, volume and thickness in reproductive failure.

In the present study, we demonstrated that vascularization of endometrium was similar for controls and women with reproductive failure, indicating a limited role of blood vessel function in implantation failure. In agreement to our study, Plaisier et al also showed no difference in vascularisation on the maternal side of implantation, namely decidualized endometrial stromal cells, in cases with missed abortions compared to controls, while they were able to demonstrated differences in receptors of growth factors in decidua parietalis and basalis at the time of missed abortion [41]. Although the importance of angiogenesis for implantation, and normal uterine and placental function is well recognized [16, 42, 43], the role of inadequate vascularization or blood flow in implantation failure has not been investigated in detail. Inconsistent results of impact of uterine blood flow in implantation failure measured by ultrasound technologies were obtained [16, 17]. For example, using ultrasound techniques [44], reduced endometrial and subendometrial 'perfusion' has been demonstrated which suggests a reduced vascularity in women with unexplained subferitility, irrespective of E2 and progesterone levels. In contrast, Ng et al. [45, 46] did not find any association between infertility and blood flow measured via ultrasound techniques. These latest results are in agreement with our studies, since we did not observe changes in blood vessel density in reproductive failure subjects. The discrepancies of results in studies cited above are likely due to different techniques (ultrasound vs. immunohistochemisty and image analysis), different population and/or study design used in these experiments. Since human studies are restricted in their design, the studies performed in human tissue are limited in their character, therefore we were only able to look at the intrinsic endometrial regulation of women with reproductive failure at the protein level. Thus, additional studies should be performed to determine mRNA and protein expression especially for genes regulating cell proliferation in endometrium from women with and without reproductive failure.

Conclusion

In summary, we demonstrate a decrease in cell proliferation in endometrial tissues from the mid-secretory phase in women with reproductive failure compared to healthy controls. However, vascularisation in endometrium was similar for the examined groups. We therefore hypothesize that adequate endometrial growth is a critical factor contributing to the success of implantation, as well as further continuation of an established pregnancy. Our results help to better understand the underlying pathomechanism of reproductive failure and may contribute to a discovery of marker(s) which can be used to predict successful vs. non-successful pregnancy.

Abbreviations

ANA:

antinuclear antibodies

ART:

assisted reproduction technique

mRNA:

messenger ribonucleic acid

C/EBPbeta:

CCAAT/enhancer binding protein (C/EBP), beta

CRABP2:

cellular retinoic acid binding protein 2

E2:

estradiol-17β

hCG:

human choriogonadotropin

IgG:

Immunglobulin G

Ki67:

proliferation marker

LH:

luteinising hormone

PBS:

phosphate buffered saline

RSA:

recurrent spontaneous abortions

RIF:

recurrent implantation failure

SG:

silver grain

SMCA:

smooth muscle cell actin.

References

  1. Farquharson RG, Jauniaux E, Exalto N: Updated and revised nomenclature for description of early pregnancy events. Hum Reprod. 2005, 20 (11): 3008-3011. 10.1093/humrep/dei167.

    Article  PubMed  Google Scholar 

  2. Margalioth EJ, Ben-Chetrit A, Gal M, Eldar-Geva T: Investigation and treatment of repeated implantation failure following IVF-ET. Hum Reprod. 2006, 21 (12): 3036-3043. 10.1093/humrep/del305.

    Article  CAS  PubMed  Google Scholar 

  3. Li TC, Makris M, Tomsu M, Tuckerman E, Laird S: Recurrent miscarriage: aetiology, management and prognosis. Hum Reprod Update. 2002, 8 (5): 463-481. 10.1093/humupd/8.5.463.

    Article  CAS  PubMed  Google Scholar 

  4. Nyboe Andersen A, Goossens V, Bhattacharya S, Ferraretti AP, Kupka MS, de Mouzon J, Nygren KG: Assisted reproductive technology and intrauterine inseminations in Europe, 2005: results generated from European registers by ESHRE: ESHRE. The European IVF Monitoring Programme (EIM), for the European Society of Human Reproduction and Embryology (ESHRE). Hum Reprod. 2009, 24 (6): 1267-1287. 10.1093/humrep/dep035.

    Article  CAS  PubMed  Google Scholar 

  5. Christiansen OB, Nielsen HS, Kolte AM: Inflammation and miscarriage. Semin Fetal Neonatal Med. 2006, 11 (5): 302-308. 10.1016/j.siny.2006.03.001.

    Article  PubMed  Google Scholar 

  6. Potdar N, Konje JC: The endocrinological basis of recurrent miscarriages. Curr Opin Obstet Gynecol. 2005, 17 (4): 424-428. 10.1097/01.gco.0000175363.20094.bd.

    Article  PubMed  Google Scholar 

  7. Tomassetti C, Meuleman C, Pexsters A, Mihalyi A, Kyama C, Simsa P, D'Hooghe TM: Endometriosis, recurrent miscarriage and implantation failure: is there an immunological link?. Reprod Biomed Online. 2006, 13 (1): 58-64. 10.1016/S1472-6483(10)62016-0.

    Article  CAS  PubMed  Google Scholar 

  8. Savaris RF, Hamilton AE, Lessey BA, Giudice LC: Endometrial gene expression in early pregnancy: lessons from human ectopic pregnancy. Reprod Sci. 2008, 15 (8): 797-816. 10.1177/1933719108317585.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Kao LC, Germeyer A, Tulac S, Lobo S, Yang JP, Taylor RN, Osteen K, Lessey BA, Giudice LC: Expression profiling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and infertility. Endocrinology. 2003, 144 (7): 2870-2881. 10.1210/en.2003-0043.

    Article  CAS  PubMed  Google Scholar 

  10. Koler M, Achache H, Tsafrir A, Smith Y, Revel A, Reich R: Disrupted gene pattern in patients with repeated in vitro fertilization (IVF) failure. Hum Reprod. 2009, 24 (10): 2541-2548. 10.1093/humrep/dep193.

    Article  CAS  PubMed  Google Scholar 

  11. Lee J, Oh J, Choi E, Park I, Han C, Kim do H, Choi BC, Kim JW, Cho C: Differentially expressed genes implicated in unexplained recurrent spontaneous abortion. Int J Biochem Cell Biol. 2007, 39 (12): 2265-2277. 10.1016/j.biocel.2007.06.012.

    Article  CAS  PubMed  Google Scholar 

  12. Tapia A, Gangi LM, Zegers-Hochschild F, Balmaceda J, Pommer R, Trejo L, Pacheco IM, Salvatierra AM, Henriquez S, Quezada M, Vargas M, Ríos M, Munroe DJ, Croxatto HB, Velasquez L: Differences in the endometrial transcript profile during the receptive period between women who were refractory to implantation and those who achieved pregnancy. Hum Reprod. 2008, 23 (2): 340-351. 10.1093/humrep/dem319.

    Article  CAS  PubMed  Google Scholar 

  13. Bagchi MK, Mantena SR, Kannan A, Bagchi IC: Control of uterine cell proliferation and differentiation by C/EBPbeta: functional implications for establishment of early pregnancy. Cell Cycle. 2006, 5 (9): 922-925. 10.4161/cc.5.9.2712.

    Article  CAS  PubMed  Google Scholar 

  14. Barnea ER: Embryo maternal dialogue: From pregnancy recognition to proliferation control. Early Pregnancy. 2001, 5 (1): 65-66.

    CAS  PubMed  Google Scholar 

  15. Maruyama T, Yoshimura Y: Molecular and cellular mechanisms for differentiation and regeneration of the uterine endometrium. Endocr J. 2008, 55 (5): 795-810. 10.1507/endocrj.K08E-067.

    Article  CAS  PubMed  Google Scholar 

  16. Alcazar JL: Three-dimensional ultrasound assessment of endometrial receptivity: a review. Reprod Biol Endocrinol. 2006, 4: 56-10.1186/1477-7827-4-56.

    Article  PubMed Central  PubMed  Google Scholar 

  17. Fanchin R: Assessing uterine receptivity in 2001: ultrasonographic glances at the new millennium. Ann N Y Acad Sci. 2001, 943: 185-202. 10.1111/j.1749-6632.2001.tb03802.x.

    Article  CAS  PubMed  Google Scholar 

  18. Jirous J, Diejomaoh ME, Al-Abdulhadi F, Boland MH, Nazar M: A comparison of the uterine and intraovarian arterial flows in nonpregnant women having a history of recurrent spontaneous miscarriage associated with antiphospholipid syndrome. Arch Gynecol Obstet. 2004, 270 (2): 74-78. 10.1007/s00404-003-0510-0.

    Article  CAS  PubMed  Google Scholar 

  19. Lisman BA, Boer K, Bleker OP, van Wely M, van Groningen K, Exalto N: Abnormal development of the vasculosyncytial membrane in early pregnancy failure. Fertil Steril. 2004, 82 (3): 654-660. 10.1016/j.fertnstert.2004.02.119.

    Article  PubMed  Google Scholar 

  20. Murray MJ, Meyer WR, Zaino RJ, Lessey BA, Novotny DB, Ireland K, Zeng D, Fritz MA: A critical analysis of the accuracy, reproducibility, and clinical utility of histologic endometrial dating in fertile women. Fertil Steril. 2004, 81 (5): 1333-1343. 10.1016/j.fertnstert.2003.11.030.

    Article  PubMed  Google Scholar 

  21. Grazul-Bilska AT, Banerjee J, Yazici I, Borowczyk E, Bilski JJ, Sharma RK, Siemionov M, Falcone T: Morphology and function of cryopreserved whole ovine ovaries after heterotopic autotransplantation. Reprod Biol Endocrinol. 2008, 6: 16-10.1186/1477-7827-6-16.

    Article  PubMed Central  PubMed  Google Scholar 

  22. Grazul-Bilska AT, Caton JS, Arndt W, Burchill K, Thorson C, Borowczyk E, Bilski JJ, Redmer DA, Reynolds LP, Vonnahme KA: Cellular proliferation and vascularization in ovine fetal ovaries: effects of undernutrition and selenium in maternal diet. Reproduction. 2009, 137 (4): 699-707. 10.1530/REP-08-0375.

    Article  CAS  PubMed  Google Scholar 

  23. Grazul-Bilska AT, Navanukraw C, Johnson ML, Vonnahme KA, Ford SP, Reynolds LP, Redmer DA: Vascularity and expression of angiogenic factors in bovine dominant follicles of the first follicular wave. J Anim Sci. 2007, 85 (8): 1914-1922. 10.2527/jas.2007-0044.

    Article  CAS  PubMed  Google Scholar 

  24. Kirk R, Experimental Design: Procedures for the Behavioral Sciences. 1982, Pacific Grove, CA, USA: Brooks/Cole, 2

    Google Scholar 

  25. Rai R, Regan L: Recurrent miscarriage. Lancet. 2006, 368 (9535): 601-611. 10.1016/S0140-6736(06)69204-0.

    Article  PubMed  Google Scholar 

  26. Jauniaux E, Burton GJ: Pathophysiology of histological changes in early pregnancy loss. Placenta. 2005, 26 (23): 114-123. 10.1016/j.placenta.2004.05.011.

    Article  CAS  PubMed  Google Scholar 

  27. Strowitzki T, Germeyer A, Popovici R, von Wolff M: The human endometrium as a fertility-determining factor. Hum Reprod Update. 2006, 12 (5): 617-630. 10.1093/humupd/dml033.

    Article  PubMed  Google Scholar 

  28. Tantbirojn P, Crum CP, Parast MM: Pathophysiology of placenta creta: the role of decidua and extravillous trophoblast. Placenta. 2008, 29 (7): 639-645. 10.1016/j.placenta.2008.04.008.

    Article  CAS  PubMed  Google Scholar 

  29. Mitselou A, Ioachim E, Kitsou E, Vougiouklakis T, Zagorianakou N, Makrydimas G, Stefanaki S, Agnantis NJ: Immunohistochemical study of apoptosis-related Bcl-2 protein and its correlation with proliferation indices (Ki67, PCNA), tumor suppressor genes (p53, pRb), the oncogene c-erbB-2, sex steroid hormone receptors and other clinicopathological features, in normal, hyperplastic and neoplastic endometrium. In Vivo. 2003, 17 (5): 469-477.

    CAS  PubMed  Google Scholar 

  30. Niklaus AL, Aubuchon M, Zapantis G, Li P, Qian H, Isaac B, Kim MY, Adel G, Pollard JW, Santoro NF: Assessment of the proliferative status of epithelial cell types in the endometrium of young and menopausal transition women. Hum Reprod. 2007, 22 (6): 1778-1788. 10.1093/humrep/dem032.

    Article  CAS  PubMed  Google Scholar 

  31. Hapangama DK, Turner MA, Drury JA, Quenby S, Hart A, Maddick M, Martin-Ruiz C, von Zglinicki T: Sustained replication in endometrium of women with endometriosis occurs without evoking a DNA damage response. Hum Reprod. 2009, 24 (3): 687-696. 10.1093/humrep/den416.

    Article  CAS  PubMed  Google Scholar 

  32. Mertens HJ, Heineman MJ, Evers JL: The expression of apoptosis-related proteins Bcl-2 and Ki67 in endometrium of ovulatory menstrual cycles. Gynecol Obstet Invest. 2002, 53 (4): 224-230. 10.1159/000064569.

    Article  CAS  PubMed  Google Scholar 

  33. Moller B, Rasmussen C, Lindblom B, Olovsson M: Expression of the angiogenic growth factors VEGF, FGF-2, EGF and their receptors in normal human endometrium during the menstrual cycle. Mol Hum Reprod. 2001, 7 (1): 65-72. 10.1093/molehr/7.1.65.

    Article  CAS  PubMed  Google Scholar 

  34. Smith SK: Angiogenesis, vascular endothelial growth factor and the endometrium. Hum Reprod Update. 1998, 4 (5): 509-519. 10.1093/humupd/4.5.509.

    Article  CAS  PubMed  Google Scholar 

  35. Punyadeera C, Verbost P, Groothuis P: Oestrogen and progestin responses in human endometrium. J Steroid Biochem Mol Biol. 2003, 84 (4): 393-410. 10.1016/S0960-0760(03)00061-X.

    Article  CAS  PubMed  Google Scholar 

  36. Carranza-Lira S, Blanquet J, Tserotas K, Calzada L: Endometrial progesterone and estradiol receptors in patients with recurrent early pregnancy loss of unknown etiology--preliminary report. Med Sci Monit. 2000, 6 (4): 759-762.

    CAS  PubMed  Google Scholar 

  37. Ilesanmi AO, Hawkins DA, Lessey BA: Immunohistochemical markers of uterine receptivity in the human endometrium. Microsc Res Tech. 1993, 25 (3): 208-222. 10.1002/jemt.1070250304.

    Article  CAS  PubMed  Google Scholar 

  38. Raga F, Bonilla-Musoles F, Casan EM, Klein O, Bonilla F: Assessment of endometrial volume by three-dimensional ultrasound prior to embryo transfer: clues to endometrial receptivity. Hum Reprod. 1999, 14 (11): 2851-2854. 10.1093/humrep/14.11.2851.

    Article  CAS  PubMed  Google Scholar 

  39. Zollner U, Zollner KP, Specketer MT, Blissing S, Muller T, Steck T, Dietl J: Endometrial volume as assessed by three-dimensional ultrasound is a predictor of pregnancy outcome after in vitro fertilization and embryo transfer. Fertil Steril. 2003, 80 (6): 1515-1517. 10.1016/j.fertnstert.2003.08.004.

    Article  PubMed  Google Scholar 

  40. Zaidi J, Campbell S, Pittrof R, Tan SL: Endometrial thickness, morphology, vascular penetration and velocimetry in predicting implantation in an in vitro fertilization program. Ultrasound Obstet Gynecol. 1995, 6 (3): 191-198. 10.1046/j.1469-0705.1995.06030191.x.

    Article  CAS  PubMed  Google Scholar 

  41. Plaisier M, Dennert I, Rost E, Koolwijk P, van Hinsbergh VW, Helmerhorst FM: Decidual vascularization and the expression of angiogenic growth factors and proteases in first trimester spontaneous abortions. Hum Reprod. 2009, 24 (1): 185-197. 10.1093/humrep/den296.

    Article  CAS  PubMed  Google Scholar 

  42. Gargett CE, Rogers PA: Human endometrial angiogenesis. Reproduction. 2001, 121 (2): 181-186. 10.1530/rep.0.1210181.

    Article  CAS  PubMed  Google Scholar 

  43. Reynolds LP, Grazul-Bilska AT, Redmer DA: Angiogenesis in the female reproductive organs: pathological implications. Int J Exp Pathol. 2002, 83 (4): 151-163. 10.1046/j.1365-2613.2002.00277.x.

    Article  PubMed Central  PubMed  Google Scholar 

  44. Raine-Fenning NJ, Campbell BK, Kendall NR, Clewes JS, Johnson IR: Endometrial and subendometrial perfusion are impaired in women with unexplained subfertility. Hum Reprod. 2004, 19 (11): 2605-2614. 10.1093/humrep/deh459.

    Article  CAS  PubMed  Google Scholar 

  45. Ng EH, Chan CC, Tang OS, Yeung WS, Ho PC: Factors affecting endometrial and subendometrial blood flow measured by three-dimensional power Doppler ultrasound during IVF treatment. Hum Reprod. 2006, 21 (4): 1062-1069. 10.1093/humrep/dei442.

    Article  PubMed  Google Scholar 

  46. Ng EH, Chan CC, Tang OS, Yeung WS, Ho PC: The role of endometrial blood flow measured by three-dimensional power Doppler ultrasound in the prediction of pregnancy during in vitro fertilization treatment. Eur J Obstet Gynecol Reprod Biol. 2007, 135 (1): 8-16. 10.1016/j.ejogrb.2007.06.006.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported in part by grant P20 RR016741from the INBRE program of the National Center for Research Resources and ND Hatch Project ND01712. Presented in part at the 42nd Annual Meeting of the Society for the Study of Reproduction, Pittsburgh, PA, USA, New Orleans, LA, July 18-22, 2009. The authors would like to thank Mr. Kim C. Kraft, Mr. Robert Weigl and Mr. James D. Kirsch for their technical assistance.

Author information

Authors and Affiliations

  1. Department of Gynecological Endocrinology and Reproductive Medicine,, University Hospital Heidelberg,, Heidelberg,, Germany

    Ariane Germeyer, Julia Jauckus & Thomas Strowitzki

  2. Department of Gynecological Endocrinology and Reproductive Medicine,, University Hospital Berne,, Berne,, Switzerland

    Michael von Wolff

  3. Department of Animal Sciences and Cell Biology Center,, North Dakota State University,, Fargo,, North Dakota,, USA

    Tanuj Sharma & Anna T Grazul-Bilska

Authors
  1. Ariane Germeyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael von Wolff
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Julia Jauckus
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thomas Strowitzki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tanuj Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anna T Grazul-Bilska
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ariane Germeyer.

Additional information

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

AG participated in designing the study, coordinated tissue collection, evaluated results and participated in writing the manuscript and was responsible for intellectual content of the study; MvW participated in patient selection, consenting the patients and tissue collection; JJ participated in tissue collection and preparation for immunohistochemistry; TS participated in patient selection and in writing of the manuscript; TS performed image analysis and prepared data for statistical analysis; ATGB participated in designing of the study, evaluated results and participated in writing the manuscript, coordinated and supervised immunohistochemical staining and image analysis. All authors read and approved the final manuscript.

Authors’ original submitted files for images

Rights and permissions

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and permissions

About this article

Cite this article

Germeyer, A., von Wolff, M., Jauckus, J. et al. Changes in cell proliferation, but not in vascularisation are characteristic for human endometrium in different reproductive failures - a pilot study. Reprod Biol Endocrinol 8, 67 (2010). https://doi.org/10.1186/1477-7827-8-67

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/1477-7827-8-67

Keywords

Reproductive Biology and Endocrinology

ISSN: 1477-7827

Contact us