Endometrial Vascular Density Research Articles

Ovarian stimulation and exogenous progesterone affect the endometrial miR-16-5p, VEGF protein expression, and angiogenesis

Angiogenesis, where vascular endothelial growth factor (VEGF) is critically involved, is an important factor in endometrial receptivity. Angio-miRNAs form a special class of microRNAs (miRNAs) that target angiogenic genes and regulate angiogenesis. Various studies have shown that ovarian stimulation and exogenous progesterone affect endometrial vascular density. The present research aimed to assess the impact of HMG/HCG and progesterone on miR-16-5p, VEGF protein expression, and angiogenesis in the mouse endometrium during the preimplantation period. Forty adult female mice were divided into four groups: 1) control, 2) ovarian stimulation (HMG and 48 h after HCG IP), 3) progesterone (progesterone IP for 3 days), 4) ovarian stimulation + progesterone (HMG and 48 h after HCG IP) + (progesterone IP for 3 days) groups.The mice were sacrificed 96 h following HCG administration. miR-16-5p, VEGF protein expression, and CD31-positive cell (Endothelial cell) density were specified.The results showed that endothelial cell density,VEGF protein, and miR-16-5p expression increased in all treatment groups, with the maximum increase belonging to the ovarian stimulation + progesterone group. This study provides evidence that ovarian stimulation and progesterone administration enhance endometrial angiogenesis through VEGF protein upregulation. Furthermore, except for miR-16-5p, other miRNAs and molecules appear to be involved in angiogenic pathways, thereby requiring further studies.

Supplemental progesterone during early pregnancy exerts divergent responses on embryonic characteristics in sows and gilts

Progesterone (P4) plays a key role in pregnancy establishment and maintenance; during early pregnancy, P4 stimulates the production and release of uterine secretions necessary for conceptus growth prior to implantation; therefore, exogenous P4 supplementation may improve embryo development. This study evaluated the effects of supplementation during early pregnancy with long-acting injectable progesterone or altrenogest on embryonic characteristics of sows and gilts. Thus, a total of 32 sows and 16 gilts were used. On day 6 of pregnancy sows and gilts were allocated to one of the following groups: non-supplemented; supplemented with 20 mg of altrenogest, orally, from days 6 to 12 of pregnancy; supplemented with 2.15 mg/kg of long-acting injectable progesterone on day 6 of pregnancy. Animals were killed on day 28 of pregnancy, and ovulation rate, embryo survival, embryo weight, crown-to-rump length, uterine glandular epithelium and endometrial vascularization were assessed. Treatments had no effect on pregnancy rate, embryo survival or endometrial vascular density (P > 0.05). Non-supplemented gilts presented larger and heavier embryos compared to gilts from supplemented groups (P < 0.05). Sows in the altrenogest group presented larger and heavier embryos compared to non-supplemented sows and sows supplemented with long-acting injectable progesterone. In conclusion, supplementation of sows and gilts with progestagen from day 6 of pregnancy can be used as a means to improve embryo survival without deleterious effects.

Equine endometrial vascular pattern changes during the estrous cycle examined by Narrow Band Imaging hysteroscopy

The aim of this study was to evaluate the uterine blood supply and endometrial vessel architecture, during the equine estrous cycle. Narrow Band Imaging (NBI) hysteroscopy was used for evaluating changes in the endometrial vasculature during the estrous cycle [six mares, d 0 (representing the day of ovulation), d 6 and 11 in four locations]. In addition, endometrial biopsy samples were used for immunodetection of markers for angiogenesis (Vascular Endothelial Growth Factor A, its receptor 2, as well as angiopoietin-2 and its receptor-tyrosine-kinase Tie2) during the estrous cycle (three mares, d 0, 5 and 10; one biopsy per mare). Detailed analysis of hysteroscopic images revealed an increase in the vascular density from estrus towards diestrus. In contrast, microscopic specimens prepared from biopsies revealed no evidence for changes in the endometrial vessel number during the estrous cycle. Studies on expression of angiogenesis markers indicated that cyclic changes in the endometrial vascular density observed by NBI-hysteroscopy were not due to formation of new vessels. It is concluded that vessels are involved in blood supply of a smaller area during diestrus, facilitating better distribution of nutrients during this phase.

Alterations in endometrial vascular density via hysteroscopy evaluated by vascular analysis software during laparoscopic myomectomy on an infertile woman with submucous myoma

Submucous myomas cause infertility and recurrent pregnancy loss. Several studies have reported successful reproductive outcomes after hysteroscopic myomectomy (HM), but the risk of postoperative intrauterine adhesion is present. We performed laparoscopic myomectomy (LM) for a submucous myoma and second look laparoscopy under observation using a hysteroscope to evaluate the alteration in endometrial vascular density during surgery using vascular analysis software.The patient was a 33-year-old nulliparous infertile woman. She had one submucous myoma of 4 cm in diameter and 50-60% penetration into the myometrium (class T:II; the European Society of Hysteroscopy classification). The surface vascular density of the submucous myoma was 68.6% before the start of surgery, decreased to 51.4% upon vasopressin injection and increased to 67.6% at the end of LM. Intraabdominal and intrauterine adhesions were not seen at second look laparoscopy. The vascular density was 70.8%, and the rate of endometrial blood flow was increased to 112.5% by comparison with the vascular density before the start of surgery. HM is a safe and effective procedure, but carries the risk of scarring the uterine cavity with uterine adhesions. We have to consider LM for women of reproductive age who have a submucous myoma with penetration >50% into the myometrium (class T:II).

A comparative study of Cyclofem® and depot medroxyprogesterone acetate (DMPA) effects on endometrial vasculature

ObjectivesThe most common reason for discontinuation of long-acting progestogen-only contraceptives is irregular bleeding following local endometrial vascular changes. To reduce unpredictable bleeding episodes among depot medroxyprogesterone acetate (DMPA) users, the...

Endometrial histology, microvascular density and caliber, and matrix metalloproteinase-3 in users of the Nestorone®-releasing contraceptive implant with and without endometrial breakthrough bleeding

Objective This descriptive study evaluated endometrial histology, microvascular density and caliber, and quantification of matrix metalloproteinase (MMP-3) expression in long-term users of the Nestorone® (NES)-releasing implant who presented or not endometrial breakthrough bleeding (BTB). Methods Endometrial biopsies were obtained from 32 healthy women with unpredictable BTB. The quantitative analysis was performed only in 20 samples. Results The mean duration of use of the implant among the 14 women with BTB was 19.6±1.0 months, and the other six women had used the implant for 17.7±2.3 months (mean±S.E.M.). Histological analysis of the endometrial tissue showed a predominance of progestogenic pattern followed by atrophic and proliferative endometrium in both groups. Mucosal breakdown and glandular pseudostratification were observed in half of the cases. Endometrial vascular density was 73.1±10.0 and 57.5±24.1 vessels/mm 2, and maximum vessel diameter was 923.3±86.0 and 1038.0±404 μm (mean±S.E.M.) in the group with and without BTB, respectively, without significance, and the rate of cells expressing MMP-3×1000 counted stromal cells was 155.8±24.8 and 127.0±19.0 (mean±S.E.M.) in both groups, respectively, without significance. Conclusions This study provides information about some endometrial aspects of women using NES in contraceptive implants. In addition, the endometrium was similar during long-term use of NES-releasing contraceptive implants in women with and without endometrial bleeding.

Effect of long-term progestin treatment on endometrial vasculature in normal cycling mice

The aim of this study was to develop a mouse model to investigate the effects of long-term progestin-only exposure on endometrial vascular structure. Normal cycling mice received Silastic implants containing either medroxyprogesterone acetate (MPA) or levonorgestrel (LNG) and were dissected after 1, 3 or 6 weeks. Endometrial vascular density increased significantly within 1 week of MPA (482 ± 40.2 vessels/mm 2) or LNG (440 ± 26.5 vessels/mm 2) treatment compared with normal cycling mice (293 ± 10.5 vessels/mm 2). MPA increased stromal cell density within 1 week of treatment (13813 ± 1450 cells/mm 2) compared with normal cycling mice (8256 ± 928 cells/mm 2). However, although LNG significantly increased stromal cell density overall, the increase did not reach significance within the individual weeks examined. There was no significant change in the ratio of vascular to stromal cell density among treated and normal cycling mice. Epithelial cell height significantly decreased within 1 week of LNG (17.6 ± 1.3 μm) treatment compared with normal cycling mice (23.5 ± 1.3 μm); epithelial cell height also decreased within 1 week of MPA treatment (16.6 ± 2.1 μm), although this did not reach statistical significance. VEGF immunostaining increased significantly in luminal epithelium after MPA or LNG treatment, and in glandular epithelium after LNG treatment. These observations are similar to those reported in human endometrium, suggesting that this mouse model may facilitate further investigations into breakthrough bleeding due to long-term progestin use.

Changes in vascular basement membrane in the endometrium of Norplant users

Progestogen-only contraception is almost invariably associated with changes in menstrual bleeding patterns. Changes in the endometrial vasculature, and in particular an increase in vascular fragility, may contribute to this bleeding. In this study, endometrial vascular density and endothelial cell basement membrane components were examined using immunohistochemistry before and after insertion of Norplant. Endometrial vascular density was increased from a mean (+/- SEM) of 189.6 +/- 7.0 vessels/mm2 during the control cycle to 253.9 +/- 80.7 vessels/mm2 at 2-13 weeks of Norplant exposure, and to 212.7 +/- 12.9 vessels/mm2 at 14-42 weeks. During the control cycle, a mean of 161.4 +/- 4.5 vessels/mm2 stained for collagen IV (85% of all vessels), while at 2-13 weeks, 144.5 +/- 13.0 vessels/mm2 stained for collagen IV (57% of all vessels) (t ratio = 2.08, P = 0.0057). By 14-42 weeks, 71% of vessels (151.0 +/- 9.8) vessels/mm2 were surrounded by collagen IV. This was not significantly different from control values (t ratio = 2.03). Endometrial vascular laminin was also reduced following Norplant insertion, from a mean of 176.0 +/- 4.2 vessels/mm2 in the control cycle (93% of vessels), to 156.3 +/- 6.7 vessels/mm2 at 2-13 weeks of exposure (57% of vessels) (t ratio = 2.08, P = 0.01). By 14-42 weeks of exposure to Norplant, 162.5 +/- 9 vessels/mm2 (76%) stained for laminin. This was not significantly different from control values (t ratio = 2.04). Endometrial vascular heparan sulphate proteoglycan (HSPG) was reduced from 58.6 +/- 3.0 vessels/mm2 during the control cycle (31% of vessels) to 43.6 +/- 5.6 vessels/mm2 (only 17% of vessels) at 2-13 weeks (t ratio = 2.08, P = 0.025). At 14-42 weeks, only 19% of vessels stained for HSPG (41.3 +/- 5.8 vessels/mm2; t ratio = 2.04, P = 0.009).

A longitudinal study of changes in endometrial microvascular density in norplant® implant users

This study aimed to investigate the effects of the subdermal levonorgestrel contraceptive implant Norplant® on endometrial vascular density at different durations of exposure, and the relationship between endometrial histology, vascular density, and bleeding patterns. A prospective controlled trial of Norplant implant users compared endometrial vascular density in biopsies taken before and after Norplant implant insertion. A total of 34 women with regular menstrual cycles requesting long-term contraception were recruited at the Sydney Centre for Reproductive Health Research, Australia. A significant increase in mean endometrial microvascular density was observed from as early as 3 weeks after insertion of Norplant implants. Vascular density was increased from a control secretory phase value of 189.6 ( 7.0 vessels/mm 2 (±SEM) to 253.80 ± 7 vessels/mm 2 at 2–13 weeks of Norplant implant exposure (t ratio = 2.08, p = 0.01) and 212.7 ± 12.9 vessels/mm 2 at 14–42 weeks of exposure (t ratio = 2.03, p = 0.02). In those with atrophic endometrium, or in whom myometrium and basalis only were found in biopsies (20 of 66, 30%), mean endometrial vascular density was increased at 273.1 ± 16.1 vessels/mm 2 compared with 210.9 ± 11.7 vessels/mm 2 in other histological groups (F ratio = 9.74, p = 0.0028). Bleeding and spotting in the previous 30 days were less common in those with this histological appearance at a mean of 4.95 days compared with 8.22 days. This is the first study to assess endometrial vascular density in the early months of Norplant implant use. The findings suggest that the endometrial vasculature is profoundly altered in the early months of Norplant implant exposure when bleeding problems are most common.

The impacts of uterine environment and fetal genotype on conceptus size and placental vascularity during late gestation in pigs.

Meishan and Yorkshire gilts were bred exclusively to Yorkshire and Meishan boars, respectively, resulting in similar Meishan x Yorkshire fetuses gestating in the uteri of both maternal breeds. Gilts of both breeds were slaughtered on d 90 and 110 of gestation, and the weight and volume of each uterine horn was determined. Fetal weights and crown-rump lengths were recorded. A section of the chorioallantoic-endometrial attachment site was collected for each conceptus and evaluated for placental and endometrial vascular density. An intact placenta was recovered for each conceptus and weighed, and its surface area was determined. Fetal weight and crown-rump length increased (P < .001) markedly between d 90 and 110 of gestation in Meishan and in Yorkshire uteri, but they were markedly less (P < .001) in Meishan than in Yorkshire uteri. Placental weight and placental surface area were reduced by 40% in Meishan, compared with Yorkshire, uteri; however, placental size did not increase between d 90 and 110 in either uterine type. Placental vascular density and associated endometrial vascular density were similar (P > .20) for conceptuses in Meishan and Yorkshire uteri on d 90 and 110 of gestation. Additionally, even though the ratio of fetal weight: placental weight (placental efficiency) increased between d 90 and 110, placental efficiency was similar for conceptuses in Meishan and Yorkshire uteri. In a previous study using straightbred Meishan or Yorkshire conceptuses gestated in either a Meishan or Yorkshire uterus, we found Meishan conceptuses had a markedly greater placental efficiency than did Yorkshire conceptuses, regardless of the type of uterus in which they were gestated. These data indicate that uterine type determines conceptus size and conceptus genotype controls placental efficiency.

The impact of either a Meishan or Yorkshire uterus on Meishan or Yorkshire fetal and placental development to days 70, 90, and 110 of gestation.

Straightbred Yorkshire (Y) conceptuses are larger than straightbred Meishan (M) conceptuses throughout gestation and at farrowing. In contrast, when Y and M conceptuses were gestated together in Y recipient females, the birth weight of M pigs was similar to that of their Y littermates. Even though placentae of M pigs remained markedly smaller than placentae of Y littermates, they were significantly more vascular. The objective of this study was to compare and contrast the changes in Y and M conceptus growth and placental-endometrial vascularity throughout late gestation in Y or M uteri. Gravid uteri were recovered at slaughter from M and Y females that were gestating either M or Y conceptuses on d 70, 90, or 110 of gestation. Uterine and conceptus measurements were recorded, and a section of the intact endometrial-placental attachment site for each conceptus was fixed, embedded, and later evaluated for placental and endometrial vascular density. Placental surface area and weight were greater (P < .001) when M or Y conceptuses were recovered from Y uteri compared with M uteri on each day of gestation examined. Further, by d 110, the surface area of Y placentae was greater (P < .001) than that of M placentae, regardless of uterine type in which they were gestated. The vascular density of M placentae and adjacent endometrium doubled (P < .05) between d 70 and 110 of gestation (3.0 and 2.5 vs 6.0 and 5.1%, respectively), with no significant increase in placental surface area. In contrast, the surface area of Y placentae doubled in size (P < .001) between d 90 and 110 of gestation, but placental and adjacent endometrial vascular density remained relatively constant, averaging 3.2 and 3.8%, respectively. These data are consistent with the premise that placental size is largely determined by the uterus in which a conceptus is gestated until approximately d 90. After d 90, fetal breed-specific mechanisms maintain optimal fetal growth. Between d 90 and term, M fetal growth depends on progressive increases in placental blood vessel density and requires no increase in placental size. In contrast, Y conceptuses seem to rely exclusively on placental growth to increase placental-endometrial surface area for nutrient exchange.