Terminal Villi Research Articles

Bioengineering approaches for patient-specific analysis of placenta structure and function

The leading cause of perinatal mortality is fetal growth restriction (FGR), defined as in utero fetal growth below the 10th percentile. Insufficient exchange of oxygen and nutrients at the maternal-fetal interface is associated with FGR. This transport occurs through the vasculature of the placenta, particularly in the terminal villi, where the vascular membranes have a large surface area and are the thinnest. Altered structure of the placenta villi is thought to contribute to decreased oxygen exchange efficiency, however, understanding how the three-dimensional microstructure and properties decrease this efficiency remains a challenge. Here, a novel, multiscale workflow is presented to quantify patient-specific biophysical properties, 3D structural features, and blood flow of the villous tissue. Namely, nanoindentation, optical coherence tomography, and ultrasound imaging were employed to measure the time-dependent material properties of placenta tissue, the 3D structure of villous tissue, and blood flow through the villi to characterize the microvasculature of the placenta at increasing length scales. Quantifying the biophysical properties, the 3D architecture, and blood flow in the villous tissue can be used to infer changes in maternal-fetal oxygen transport at the villous membrane. Overall, this multiscale understanding will advance knowledge of how microvascular changes in the placenta ultimately lead to FGR, opening opportunities for diagnosis and intervention.

Unique Ultrastructural Alterations in the Placenta Associated With Macrosomia Induced by Gestational Diabetes Mellitus

Abstract Objective To investigate the morphological and ultrastructural alterations in placentas from pregnancies with gestational diabetes mellitus (GDM)–induced macrosomia, term nondiabetic macrosomia, and normal pregnancies. Methods Sixty full-term placentas were collected, and clinical data along with informed consent were obtained from pregnant women who underwent regular visit checks and delivered their newborns in Northwest Women’s and Children’s Hospital between May and December 2022. Placentas were divided into three equal groups: normal pregnancy (control group), nondiabetic macrosomia group, and macrosomia complicated with GDM (diabetic macrosomia) group. Gross morphological data of placentas were recorded, and placental samples were processed for examination of ultrastructural and stereological changes using transmission electron microscopy. Analysis of variance and chi-squared test were used to examine the differences among the three groups for continuous and categorical variables, respectively. Results The baseline characteristics of mothers and neonates did not differ across the three groups, except for a significantly higher birth weight in the diabetic macrosomia group (4172.00 ± 151.20 vs. 3192.00 ± 328.70, P < 0.001) and nondiabetic macrosomia group (4138.00 ± 115.20 vs. 3192.00 ± 328.70, P < 0.001) compared with control group. Examination of the placentas revealed that placental weight was also highest in the diabetic macrosomia group compared with control group (810.00 ± 15.81 vs. 490.00 ± 51.48, P < 0.001) and nondiabetic macrosomia group (810.00 ± 15.81 vs. 684.00 ± 62.69, P < 0.001), but the ratio of neonatal birth weight to placental weight (BW/PW) was significantly lower in the diabetic macrosomia group compared with that in the control group (5.15 ± 0.19 vs. 6.54 ± 0.63, P < 0.001) and nondiabetic macrosomia group (5.15 ± 0.19 vs. 6.09 ± 0.52, P < 0.001) group. In contrast, the BW/PW ratio in nondiabetic macrosomia did not differ significantly from that in the control group. Distinct ultrastructural changes in terminal villi and stereological alterations in microvilli were observed in the diabetic macrosomia group, including changes in the appearance of cytoplasmic organelles and the fetal capillary endothelium and thickness of the vasculo-syncytial membrane and basal membrane. Conclusion Significant ultrastructural and stereological alterations were discovered in the placentas from pregnant women with macrosomia induced by GDM. These alterations may be the response of the placenta to the hyperglycemia condition encountered during pregnancies complicated with GDM.

HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL FEATURES OF DIFFERENTIATED TROPHOBLAST IN CHORIONIC VILLI OF THE PLACENTA IN PRETERM LABOR

The importance of studying the histochemical and immunohistochemical features of the diff erentiated trophoblast in chorionic villi of the placenta in preterm labor is determined by the threatening nature of this condition for both mother and fetus, as well as its high prevalence and serious consequences for the health of both. The understanding of the damage to the trophoblast of chorionic villi in the placenta can be expanded by histochemical and immunohistochemical methods, which allow the assessment of the concentration of specifi c marker molecules in one way or another.Objective. To determine certain histochemical and immunohistochemical characteristics of proteins in the trophoblast of theintermediate and terminal chorionic villi of the placenta in preterm labor.Material and Methods. The obtained material (30 placentas from preterm deliveries and 30 placentas from normalpregnancies) was fi xed for 20-24 hours in 10 % neutral formalin solution buff ered in Lilly’s phosphate buff er. After tissue removal, the placental tissue was dehydrated in an ascending ethanol series and embedded in paraffi n at a temperature of approximately 58 °C. Serial histologic sections were cut at 5.0 μm thickness using an MS-2 sliding microtome. After deparaffi nization, histological sections were stained with hematoxylin and eosin, histochemical methods for total protein with bromophenol blue according to Bonhéme, and immunohistochemical techniques according to the manufacturer’s protocols (Dako, Denmark). In particular, immunohistochemical reactions were performed with monoclonal antibodies against trophoblast hormone- placental lactogen and placental alkaline phosphatase. Visualization of primary antibodies was performed using the Dako polymer visualization system with diaminobenzidine as chromogen (resulting in brown staining of the sites of studied antigens). In addition to the descriptive method of histopathologic research, computer morphometry of digital microphotographs of histologic sections was performed using a Delta Optical Evolution 100 microscope and an Olympus SP550UZ digital camera. Digital copies of the images were processed using a legitimate copy of the ImageJ v1.52f computer program developed for histometric studies (National Institutes of Health, USA). Specifi cally, the evaluation of staining intensity (optical density) was performed on digital microphotographs using the method of computer microdensitometry. For this purpose, a microprobe method was used to obtain a computer brightness value in an 8-bit analysis system with 256 gray levels – from black (0) to white (255).The obtained values were then transformed into relative optical density values (r.OD) by logarithmic transformation (naturallogarithm method). The relative optical density value ranges from 0 (absolute transparency of the object) to 1 (absolute opacity of the object). The obtained digital data were processed using statistical analysis methods. A legitimate copy of the statistical analysis computer program PAST v4.14 was used, with a preliminary check for normal distribution using the Shapiro- Wilk test. Since, according to this test, the hypothesis of normal distribution was not rejected for the statistical samples studied (at p=0.05), parametric methods of statistical analysis were applied: calculation of the mean and its standard error, Student’s t-test (two-tailed, unpaired). In addition, the non-parametric Mann- Whitney test was used for the reliability of the conclusions. The research was conducted as part of the scientifi c research project «Preservation and Restoration of Reproductive Health in Women and Girls with Obstetric and Gynecological Pathology» at the Department of Obstetrics and Gynecology of Bukovinian State Medical University (state registration number 0121U110020, duration 2021-2025).Results. Microscopic examination of histological sections stained with hematoxylin and eosin did not reveal any diff erencesin the structure of chorionic villus trophoblast in the placenta of preterm labor compared to normal pregnancy. However,histochemical and immunohistochemical methods of investigation revealed a number of features in the trophoblast of chorionic villi, where fundamental events related to substance exchange occur – intermediate immature, intermediate mature, terminal villi, including terminal «specialized villi».Conclusions. According to the obtained histochemical and immunohistochemical data, in preterm labor, compared to normalpregnancy, no changes are observed in the trophoblast of intermediate immature villi, while in intermediate mature and terminal villi, there is a decrease in histochemical staining for total protein and immunohistochemical staining for specifi c trophoblast proteins – placental lactogen hormone and placental alkaline phosphatase enzyme.

PATHOMORPHOLOGICAL CHANGES OF THE PLACENTA IN THE ACUTE PERIOD OF COVID-19 IN PREGNANT WOMEN

Introduction. The barrier function of the mature placenta ensures the birth of PCR-negative infants in women during the acute period of coronavirus disease (COVID-19). This was explained by a decrease in the level of activity and concentration of angiotensin-converting enzyme type II protein (ACE2), which is a receptor for the coronavirus SARS-CoV-2 in the placenta. We conducted this study in order to detect compensatory changes in the placenta during COVID-19 in a pregnant woman. The aim of the study: To determine pathomorphological changes of the placenta in the acute period of COVID-19 in a pregnant woman. Materials and methods. The placentas of women with full-term pregnancy who were having COVID-19 confirmed by a positive PCR test (determination of SARS-CoV-2 RNA) during childbirth (n=32) were studied. The duration of the post-COVID interval (time from the diagnosis of COVID-19 to delivery) equaled 1–4 weeks. Macroscopic, organometric, microscopic, statistical, and immunohistochemical (with determination of CD34 monoclonal antibody expression) research methods were used. To determine the percentage of the studied structures that differed in color after histological staining, archival photos (×400) were uploaded to ONLINE JPG TOOLS. Results. Delivery dates: 37 to 42 weeks of gestation (median 40). COVID-19 occurred in the period 34 to 40 weeks of gestation (median 36). Course of coronavirus disease in pregnant women: 10 – mild, 18 – medium-severe, severe condition with bilateral polysegmental pneumonia – 4 women. Evaluation of newborns according to the Apgar score: 21 children (65.6%) – 8–9 points, 9 cases (28.1%) – 7–8 points, and one observation – 3–4 points and 6–7 points. No significant difference was found between the clinical manifestations of COVID-19 or the severity of the course of the disease in the pregnant woman and the morphological changes of the placenta. Microscopic examination of the placenta revealed placentitis: chorioamnionitis – 93.8% (95%CI: 82.4%–99.5%), basal deciduitis – 100% (95%CI: 94.2%–100%), villitis and intervillositis – 28.1% (95%CI: 13.7%–45.4%). The degree of manifestations of chorioamnionitis: (+) and (++) was in 34.4% (95% CI: 18.6%–52.1%), basal deciduitis (+) – 15.6% (95% CI: 5 %–30.6%), p1-2=0.08; and (++) in 28.1% (95% CI: 13.7%–45.4%). Manifestation of chorioamnionitis in (+++) and (++++) in 11.8% (95% CI: 3.2%–26.6%), basal deciduitis (+++) in 28.1% (95 % CI: 13.7%–45.4%) and (++++) in 15.6% (95% CI: 5%–30.6%). The expression of manifestations of basal deciduitis (+++++) was found in 11.8% (95% CI: 3.2%–26.6%), p1-2=0.04. The degree of severity of inflammatory changes of the maternal surface of the placenta from (+++) to (+++++) with minor similar changes in the amniotic membranes (+) and (++) confirmed the vertical route of infection. Edema of the stroma of the terminal villi was detected in 90.6% (95% CI: 77.7%–98.3%) and caused a narrowing of the lumen of the capillaries of the terminal villi of the chorion, and a decrease in the percentage of the intervillous space – 26.0 [20.7; 33.8] vs. 44.9 [40.5; 49.6] as a normal value; p = 0.0005. Percentages in a histological section of the terminal villus: stroma – 72.3 [65.8; 78.7], blood vessels – 29.0 [25.6; 34.2] vs. normal indicators: 32.1 [26.2; 39.6] and 67.8 [60.3; 73.7], respectively; p = 0.0039. Conclusions. Maternal pathomorphological changes in full-term pregnancy in the acute period of COVID-19 were caused by the exudation phase of the inflammatory process, which explained the manifestations of placentitis, swelling of the stroma of terminal villi with narrowing of the lumen of capillaries, dyscirculatory disorders. Live birth with high scores on the Apgar scale was ensured by compensatory changes of the mature placenta: an increase in the number of syncytial nodules, the locality of villitis and intervillositis, as mechanisms of increasing the intervillous space to ensure adequate perfusion.

Modeling of human placental capillaries by using a unique 3d reconstruction program

Purpose: Visualization of the three-dimensional structures is performed by examining two-dimensional sections in histology. It is not always possible to comprehend structure-function integrity. Therefore, it was planned to obtain three-dimensional (3D) models of the capillaries in the placenta terminal villus by using the program developed in the Başkent University’s laboratory and used in studies in different disciplines. The program’s suitability for histology studies will also be assessed.
 Materials and Methods: A term placenta tissue sample was obtained from a 33-year-old healthy woman. Following the plastic embedding procedure, semi-thin serial sections were taken from these blocks. The sections were stained using toluidine blue and examined and photographed by using a light microscope. The 3D modeling procedure was conducted on the light micrographs of serial sections in order to reconstruct villous capillaries. 
 Results: 3D images were obtained from two-dimensional light micrographs by using the same program. It was determined that the vessels running parallel to each other branched harmoniously, forming a capillary network in the terminal villus. Additionally, challenges encountered during the utilization of this program in histological studies are also presented in this study. 
 Conclusion: Fetal blood reaches every part of the terminal villus through the branching capillary network. This feature would increase the surface area of vasculo-syncytial membrane and facilitate feto-maternal exchange. This can be clearly visualized by using 3D models. Three-dimensional modeling helps to understand the structure of all organs and tissues. Given that the current program is freely accessible, it will not incur additional costs for researchers and is open to further improvement by the programmer (erhankiziltan@gmail.com).

Results of the morphological and immunohistochemical study of the consequences in premature birth and premature rupture of the membrane

The article presents the results of a morphological and immunohistochemical study of the afterbirths of women with miscarriages in cases of premature rupture of membranes (PRPO).The aim: to study the morphological and immunohistochemical features of placentas in premature births and premature rupture of membranes and to conduct a comparative analysis of the results depending on the gestational period.Material and methods. A morphological and immunohistochemical study of the litters of 250 pregnant women (the main group) with spontaneous premature births at 22-36 weeks of gestation and premature rupture of the membranes, which took place on the basis of the Department for Pregnant Women with Obstetric Pathology of the PAG Institute named after Acad. OHM. Lukyanova National Academy of Sciences of Ukraine" for the period from 2018 to 2020. The women of the main group are divided into three groups depending on the date of delivery - the 1st group includes patients who gave birth at 22-28 weeks of pregnancy, patients of the 2nd group who gave birth at 29-33 weeks, and to the 3rd group - those who gave birth at 34-36 weeks of pregnancy. Placentas of pregnant women of the main group were examined using morphological (histological method - staining with hematoxylin and eosin and van Gieson) and immunohistochemical methods (indirect streptavidin-peroxidase method for determining the expression level of progesterone receptors - RP).Results and discussion. Premature maturation of chorionic villi was found in 40 % of placentas of women in the 1st group 1, in 20 % – blood circulation disorders and the presence of acute inflammatory infiltration in the fetal membranes, decidual and chorionic membranes, 10 % – pathological immaturity of the placenta according to the variant of chaotic sclerosed villi. During the immunohistochemical study of RP, an unexpressed reaction was found in the epithelium and stromal cells of the stem, intermediate and terminal villi – 1 point; in fetal membranes and extra-villous cytotrophoblast – 2 points; in the endothelium of vessels – 0–1 point. In the placentas of women in the 2nd and 3nd group premature maturation of villi was found in 70 % of cases, pseudo-infarcts of villi which are embedded in fibrinoid occupying a significant area – 45 %, afunctional zones – 55 %, blood circulation disorders – 35 %, pathological immaturity placenta according to the variant of chaotic, sclerosed villi – 20 %, acute inflammatory infiltration in the fetal membranes – 15 %. Immunohistochemical analysis of the placental barrier revealed the highest expression of RP in the nuclei of decidual cell which belong to the maternal structure. Of note is the presence of progesterone receptor expression in stem villi, fibroblasts, and vessel wall nuclei. Minimal or absent expression was determined in the nuclei of other villous structures and their vessels. Conclusions. It has been proven that the receptivity of progesterone has its own characteristics in placental structures depending on the gestation period, which is an important factor in the choice of management for the prevention and reduction of perinatal losses for this contingent of pregnant women. The conducted studies make it possible to formulate a clear concept of pathogenetically justified therapy for miscarriage.

The balance between fetal oxytocin and placental leucine aminopeptidase (P-LAP) controls human uterine contraction around labor onset.

A fetal pituitary hormone, oxytocin which causes uterine contractions, increases throughout gestation, and its increase reaches 10-fold from week 32 afterward. Oxytocin is, on the other hand, degraded by placental leucine aminopeptidase (P-LAP) which exists in both terminal villi and maternal blood. Maternal blood P-LAP increases with advancing gestation under the control of non-genomic effects of progesterone, which is also produced from the placenta. Progesterone is converted to estrogen by CYP17A1 localized in the fetal adrenal gland and placenta at term. The higher oxytocin concentrations in the fetus than in the mother demonstrate not only fetal oxytocin production but also its degradation and/or inhibition of leakage from fetus to mother by P-LAP. Until labor onset, the pregnant uterus is quiescent possibly due to the balance between increasing fetal oxytocin and P-LAP under control of progesterone. A close correlation exists between the feto-placental and maternal units in the placental circulation, although the blood in the two circulations does not necessarily mix. Fetal maturation results in progesterone withdrawal via the CYP17A1 activation accompanied with fetal oxytocin increase. Contribution of fetal oxytocin to labor onset has been acknowledged through the recognition that the effect of fetal oxytocin in the maternal blood is strictly regulated by its degradation by P-LAP under the control of non-genomic effects of progesterone. In all senses, the fetus necessarily takes the initiative in labor onset.

Sex-specific impact of maternal obesity on fetal placental macrophages and cord blood triglycerides

IntroductionMaternal obesity is associated with increased risk of offspring obesity and cardiometabolic disease. Altered fetoplacental immune programming is a potential candidate mechanism. Differences in fetal placental macrophages, or Hofbauer cells (HBCs), have been observed in maternal obesity, and lipid metabolism is a key function of resident macrophages that may be deranged in inflammation/immune activation. We sought to test the following hypotheses: 1) maternal obesity is associated with altered HBC density and phenotype in the term placenta and 2) obesity-associated HBC changes are associated with altered placental lipid transport to the fetus. The impact of fetal sex was evaluated in all experiments. MethodsWe quantified the density and morphology of CD163-and CD68-positive HBCs in placental villi in 34 full-term pregnancies undergoing cesarean delivery (N = 15, maternal BMI ≥30 kg/m2; N = 19, BMI <30 kg/m2). Antibody-positive cells in terminal villi were detected and cell size and circularity analyzed using a semi-automated method for thresholding of bright-field microscopy images (ImageJ). Placental expression of lipid transporter genes was quantified using RTqPCR, and cord plasma triglycerides (TGs) were profiled using modified Wahlefeld method. The impact of maternal obesity and fetal sex on HBC features, lipid transporters, and cord TGs were evaluated by two-way ANOVA. Spearman correlations of cord TGs, HBC metrics and gene expression levels were calculated. ResultsMaternal obesity was associated with significantly increased density of HBCs, with male placentas most affected (fetal sex by maternal obesity interaction p = 0.04). CD163+ HBCs were larger and rounder in obesity-exposed male placentas. Sexually dimorphic expression of placental FATP4, FATP6, FABPPM, AMPKB1 and AMPKG and cord TGs was noted in maternal obesity, such that levels were higher in males and lower in females relative to sex-matched controls. Cord TGs were positively correlated with HBC density and FATP1 expression. DiscussionMaternal obesity is associated with sex-specific alterations in HBC density and placental lipid transporter expression, which may impact umbilical cord blood TG levels and offspring cardiometabolic programming.

Spatial transcriptomics of human placentas reveal distinct RNA patterns associated with morphology and preeclampsia

Spatial transcriptomics (ST) maps RNA level patterns within a tissue. This technology has not been previously applied to human placental tissue. We demonstrate analysis of human placental samples with ST. Unsupervised clustering revealed that distinct RNA patterns were found corresponding to different morphological structures. Additionally, when focusing upon terminal villi and hemoglobin associated structures, RNA levels differed between placentas from full term healthy pregnancies and those complicated by preeclampsia. The results from this study can provide a benchmark for future ST studies in placenta.

Study of histopathological changes and the levels of TNF-α in Preterm Preeclamptic women

Preeclampsia (PE) is a significant contributor to maternal morbidity and mortality that manifests as new-onset hypertension after 20 weeks of pregnancy with signs of uteroplacental or maternal organ dysfunction or proteinuria. This condition is linked to intrauterine growth restriction, preterm birth, placental abruption, fetal distress, and fetal death in utero. The findings of the recent study can be summed up as follows: Hematoxylin and eosin staining of placenta tissues revealed a significant area of calcification in the villous stroma. Additionally, increased collagen fibers around blood vessels and varying degrees of mononuclear cell infiltration are caused by capillary and vein congestion. The Masson's Trichrome stain revealed deposited fibrin between decidual cells and extra-villous cytotrophoblasts, which were surrounded by mature intermediate and terminal villi. Congestion of veins with increased fibrin deposits around veins and arterial vessels with increased fibrocytes When the data was statistically analyzed, it was discovered that PE pregnant women had significantly p&lt;0.05 higher levels of serum TNF- α , than normotensive women ( NT ). Finally, the study found a link between preterm PE and women's pregnancy, as well as histopathologic and immunological parameters in preterm PE and NT women.&#x0D;

Correlation Between Placental Histomorphology and Umbilical Cord Coiling Index in Preeclampsia

BACKGROUND&#x0D; Preeclampsia is a common hypertensive disorder of pregnancy and one of the leading causes of maternal health complications and death globally. Both preeclampsia and abnormal umbilical coiling have been reported to alter the structure of the placenta significantly. Cases of preeclampsia and either of the two abnormal coiling states; hypercoiling or hypocoiling, occurring in the same pregnancy have been documented. The type of abnormal coiling associated with preeclampsia varies in different populations. The extent of changes in the placenta in preeclampsia coupled with either of the two is unknown. The current study aimed at determining which of the coiling state significantly alters the structure of the placentae of mothers with preeclampsia and therefore, predisposes them to more adverse perinatal outcomes.&#x0D; &#x0D; METHODs&#x0D; An unmatched case-control study was carried out in Kenyatta National Hospital labor ward and maternity theatre. Seventy (70) placentae from mothers with diagnosis of preeclampsia (cases) and mothers without complicated pregnancy (controls) were collected. The length of the cord and the number of coils was measured to calculate the coiling index. A gross placental examination was carried out followed by biopsy and tissue processing for light microscopy. Slides were photographed and several histological features analyzed in the terminal villi. Data from this study was input into SPSS (Version 21.0) where mean values, standard deviations, and frequency tables were obtained. Appropriate parametric and non-parametric tests were used where needed. A P-value of &lt;0.05 was considered statistically significant.&#x0D; &#x0D; RESULTS&#x0D; In the current study, the mean umbilical cord coiling index (UCI) in cases and controls was 0.40 ± 0.18 and 0.22 ± 0.08 coils/cm respectively. Cases had a significantly greater UCI than controls (P = &lt; 0.001). All placentae in the case hypocoiled subdivision had infarctions and were meconium stained. The mean placental weight was 519 g and 578 g respectively in cases and controls and the difference was found to be significant (p = 0.031). In cases and controls, the gross placental volume was 542 cm3 and 638 cm3, respectively. There was a significant difference in placental volume between cases and controls, and between subdivisions (p = &lt; 0.001 and 0.028 respectively). The case hypocoiled subdivision had the highest absolute volume of villous stroma, fibrin deposition, syncytial knots and the lowest fetal capillary surface area.&#x0D; &#x0D; CONCLUSION&#x0D; Hypocoiling was associated with alteration of the structure of the placenta in preeclampsia more than it was observed in hypercoiling. Early detection of hypocoiling using ultrasound in pregnancies complicated with preeclampsia may help clinicians in identification of cases that warrant close antenatal and intrapartum monitoring thus reducing adverse perinatal outcomes.&#x0D;

Utility of the Movat pentachrome stain technique in the microanatomical analysis of the human placenta

The efficacy and utility of pentachrome staining has been demonstrated in many studies on diverse human body tissues. Movat pentachrome technique is used for multicolor staining of tissue sections and vascular and stroma visualization. So far, the utility of this method for microanatomical evaluation of placental structures has not been demonstrated. The aim of this study is to evaluate the image of normal placenta and to develop some reference images for future evaluation of pathological tissues based on this technique. Material and method. The study was carried out on 21 paraffin slices taken from seven mature human placenta of single pregnant women without significant pathology and completed with a planned, elective Caesarean section. All paraffin slices were used for preparation of tissue microarrays and then performing a histochemical staining (HE and Movat). Results. On the basis of the collected material, microscopic analysis enabled the identification of normal placental villi - terminal villi,mature intermediate villi and stem villi. Moreover the maternal part of placenta was visualized. It is worth emphasizing that in each case not only the trophoblast but also the stroma structures were visualized. Conclusions. This study proved the effectiveness and usefulness of Movat imaging of placenta especially in visualization of the stroma.

Combination of Ursodeoxycholic Acid and Glutathione Improves Intestinal Morphology in Cholestasis by Downregulating TNF-α Expression

BACKGROUND: Cholestasis caused by obstruction of the common bile duct and may developed gut-derived sepsis due to reactive oxygen species (ROS) accumulation. Ursodeoxycholic acid (UDCA) and glutathione are widely known for their antioxidant properties, that might be beneficial against ROS. However, the effects of UDCA-glutathione combination against ROS have not been well elucidated in previous studies. Thus, this study was conducted to evaluate tumor necrosis factor (TNF)-α level and height of terminal ileal mucosal villus after UDCA-glutathione administration in cholestasis rat model.METHODS: Twenty-eight male Sprague Dawley rats were randomly grouped into four treatment groups, each group consisted of seven rats that had previously undergone bile duct ligation. Three groups received treatment of UDCA-glutathione combination on stratified dose, while the other one only received UDCA. Each treatment was given for 21 days. Ileal samples were collected from the rats and stained with mouse anti TNF-a antibody and hematoxylin-eosin (HE). Immunohistochemistry and histopathological examination were done using microscope and then calculated with ImageJ.RESULTS: The combination of UDCA and glutathione treatment decreased the TNF-α expression (p&lt;0.05) compared to UDCA only group, particularly in group that received 20 mg UDCA and 15 mg glutathione supplementation (p&lt;0.05) and group that received 30 mg UDCA and 20 mg glutathione supplementation (p&lt;0.05). The height of the mucosa villous was higher in the UDCA-glutathione combination groups for all the three dosage variations given (p&lt;0.05) compared to UDCA only group.CONCLUSION: UDCA-glutathione combination downregulates TNF-α expression and improves ileum mucosal villus height in cholestasis.KEYWORDS: cholestasis, glutathione, intestinal villus height, TNF-α, UDCA

Immunohistochemical features of expression of progesterone receptors of placental structures in premature birth

The objective: to study the immunohistochemical features of placenta in women with premature birth in 28–33 weeks of pregnancy. Materials and methods. 120 pregnant women (the main group) who were hospitalized and delivered in the Kyiv Perinatal Center during 2018–2020 at a gestation period of 23–33 weeks were examined. The women of the main group were divided into two subgroups depending on the term of birth – the 1st group included patients who delivered at 23–28 weeks of pregnancy, the patients of the 2nd group had labors at 29–33 weeks. Placentas of women in the main group were examined using morphological (histological method – staining with hematoxylin and eosin and van Gieson) and immunohistochemical methods (indirect streptavidin-peroxidase method for determining the expression level of progesterone receptors – PR). Results. Premature maturation of chorionic villi was found in 40 % of placentas of women in the 1st group 1, in 20 % – blood circulation disorders and the presence of acute inflammatory infiltration in the fetal membranes, decidual and chorionic membranes, 10 % – pathological immaturity of the placenta according to the variant of chaotic sclerosed villi. During the immunohistochemical study of RP, an unexpressed reaction was found in the epithelium and stromal cells of stem, intermediate and terminal villi – 1 point; in fetal membranes and extravillous cytotrophoblast – 2 points; in the endothelium of vessels – 0–1 point. In the placentas of women in the 2nd group premature maturation of villi was found in 70 % of cases, pseudoinfarcts of villi which are embedded in fibrinoid occupying a significant area – 45 %, afunctional zones – 55 %, blood circulation disorders – 35 %, pathological immaturity placenta according to the variant of chaotic, sclerosed villi – 20 %, acute inflammatory infiltration in the fetal membranes – 15 %. Immunohistochemical analysis of the placental barrier revealed the highest expression of RP in the nuclei of decidual cell which belong to the maternal structure. It should be noted the presence of expression of progesterone receptors in stem villi, fibroblasts and nuclei of the vessel wall. Minimal or absent expression was determined in the nuclei of other villous structures and their vessels. Conclusions. It was established that the receptivity of progesterone has its own characteristics in placental structures depending on the gestation period, which is an important factor in the choice of management for the prevention and reduction of perinatal losses for this contingent of pregnant women.

The effect of gestataional hypertension on the ultrastructural structure of the placenta

The objective: to study pathological changes and compensatory-adaptive reactions in the structural and ultrastructural components of the placental barrier in pregnant women with gestational arterial hypertension, who received magnesium preparations. Materials and methods. 100 patients with gestational hypertension were examined. The presence of magnesium deficiency in these women was determined by the results of our questionnaires.Pathomorphological and electronic microscopic examination of 32 placentas of patients with gestational hypertension was carried out. 12 placentas of them were from pregnant women who received the proposed management of pregnancy, which included magnesium drugs (I group), and 11 placentas – from patients with risk of preeclampsia, who were observed with the use of standard approaches of pregnancy for women with the risk of preeclampsia (II group). Control group included the results of pathomorphological and electronic microscopic study of 9 placentas from healthy women.Results. Placental of women with gestational hypertension have structural features, which is manifested by the mosaic change of placental barrier in the form of uneven circulatory disorders with hemorrhages and stasis (at the ultrastructural level) in the fetal microvessels of the villi, reducing the placental functional area (rapprochement of the villi), presence of stromal sclerosis villi, foci of pathological immaturity, increase in calcium salts (mineral dystrophy) on the background of compensatory reactions. Electronic microscopic study of the placentas in the I group revealed an increase in the compensatory-adaptive reactions of the placenta – thinning of the placental barrier, activation of cytotrophoblast cells in the placental barrier, increase the number of syncytiocapillary membranes in terminal villi with syncytiotrophoblast organic activation, increase in the number of ribosomes, the appearance of orthodox configuration of mitochondria, increasing the number of capillary membranes and thinning of syncytia, reducing the number of sludge phenomena in microvessels, which leads to an increase in the possibility of nutrient transfer from mother to fetus, etc.Conclusions. The use of the proposed therapy, which involves magnesium, diosmin and others substances, leads to a significant decrease in the frequency of detection of pathological changes in structural and ultrastructural components of the placental barrier and increased compensatory-adaptive reactions which can be considered the result of the positive impact of the proposed therapy on the placental ultrastructural elements in women with risk of preeclampsia.

Histopathological changes in the placentas of mothers with a history of coronavirus disease 2019 infection in pregnancy- A comparative cross-sectional study

This study aimed to evaluate histopathological changes in the placentas of mothers who had contracted COVID-19 during pregnancy. This prospective study involved the histopathological assessment of two groups of placentas submitted for evaluation to the Department of Histopathology at Khoula Hospital, Oman. The first group consisted of 48 placentas derived from COVID-19-positive pregnant women delivered at the centre between March 2020 and March 2021. The control group consisted of an additional 48 placentas derived from asymptomatic mothers who were not tested for COVID-19 but were assumed to be negative. All placentas underwent gross and microscopic histopathologic examination. Placental lesions were classified according to the Amsterdam system. There was a significantly higher frequency of fibrin thrombi at the terminal villi in placentas derived from the COVID-19-positive group compared to the control group (72.9% versus 0%; p-value = 0.001). Also, villous hypoperfusion was significantly more common in COVID-19-positive placentas than in controls (16.7% versus 0%; p-value = 0.006). However, no significant differences between the two groups were noted with regards to the frequency of other histopathologic features, including decidual vasculopathy, chorioamnionitis, funisitis, intervillositis, perivillous fibrin deposition, and infarction.

Maternal obesity and placental pathology in correlation with adverse pregnancy outcome.

Maternal obesity is associated with increased maternal and fetal morbidity and mortality, with an increased risk of gestational diabetes mellitus (GDM) and preeclampsia (PE). This prospective study histopathologically analyzes the placentas obtained from 34 pregnant obese women studied between October 2016 and May 2020. The 10 cases of term placentas from obese pregnancies with GDM and the 12 cases with PE were examined by the Hematoxylin-Eosin (HE), Masson's trichrome (MT) and Periodic Acid-Schiff-Hematoxylin (PAS-H) classical stainings, and by the immunohistochemical evaluation and compared to placentae from uncomplicated term obese pregnancies (12 cases). We did not meet placental histopathological (HP) abnormalities that we could classify as characteristic only for the state of obese pregnancy, but we did find placental changes associated with PE and GDM, in the context of obese pregnancy. In the case of association with PE, there were common lesions, manifested by intra- and perivillous fibrinoid deposition, calcification, and placental infarction area, to which were added numerous syncytial knots. In the case of obese pregnancy associated with GDM, we found, in addition to common placental lesions of obesity, intravillositary vascular edema and in the terminal villi appearing chorangiosis. This study revealed a number of HP changes that occur in maternal obesity, even in uncomplicated obese pregnancies. A characteristic of obese pregnancies associated with PE was the presence of numerous syncytial knots, and in obese pregnancies associated with GDM, the most common HP lesion was placental chorangiosis. Certainly, we cannot conclude that these HP lesions are specific to a particular pathology, but they belong primarily to the status of maternal obesity.

Morphological and histological variations of human placenta in pregnancies affected with preeclampsia and gestational diabetes – A review

The placenta is a transitory organ that is situated between the mother and the foetus. This vital organ which has nutritional, endocrine and immunologic functions so as to maintain pregnancy and promote fetal development. Several factors are related to the proper functioning of the placenta including foetal and maternal blood flow, appropriate nutrients, expression and function of receptors and transporters, and also the morphology of the placenta itself. Gestational hypertension and gestational diabetes are among the most frequent obstetric disorders during pregnancy. Pregnancies complicated with one or both of these disorders are also associated with adverse consequences for the mother and infant (both acute and long-term). Preeclampsia being one of the toxemias of pregnancy is a condition of increased blood pressure accompanied by proteinuria, edema or both. Various changes like increased syncytial knots, cytotrophoblastic cell proliferation, villous stromal fibrosis and fibrinoid necrosis was observed in placenta from preeclamptic cases. Diabetes mellitus can be defined as a chronic disorder marked by high blood glucose levels and a disruption in fat and protein metabolism. In pregnancies affected by diabetes mellitus also several changes occur like placental size, increased glycogen deposits, increased volume of intervillous space and terminal villi, fibrinoid areas, and glycogen deposits compared to others.All these changes may result in functional changes in this organ thus limiting the wellbeing of the developing foetus. Therefore, this review as a whole gives an overview of morphological changes at macroscopic and histological levels seen in the placenta from gestational diabetes and preeclampsia.

Comparative Characteristics of Sialoglycans Expression Disorders in the Placental Barrier Structures in Preeclampsia and Fetal Growth Restriction.

We compared the expression profiles of α2,3- and α2,6-sialoglycans in the glycocalyx of the placental barrier structures in early and late forms of preeclampsia and fetal growth restriction using the method of lectin histochemistry. It was found that the expression of α2,3-sialoglycans in the syncytiotrophoblast and fetal endothelium of the terminal villi of the placenta was reduced in preeclampsia in comparison with normal placenta and, on the contrary, was increased in fetal growth restriction. Significant differences were found in both clinical phenotypes of preeclampsia and fetal growth restriction. Changes in the expression pattern of α2,6-sialoglycans in the endothelium of terminal villi were more pronounced than in syncytiotrophoblast. In early and late-onset preeclampsia, a significant increase in the expression of α2,6-sialoglycans was revealed only in the fetal endothelium; in early fetal growth restriction, the expression of α2,6-sialoglycans was reduced in the endothelium, but increased in syncytiotrophoblast in late fetal growth restriction. The features of the expression of sialoglycans in structures of the placental barrier in preeclampsia and fetal growth restriction were revealed, which may indicate the pathogenetic involvement of sialoglycans in the inflammatory activation cascade in fetal growth restriction, and in preeclampsia, apparently, they are associated with impaired fetal tolerance.

CAN PLACENTAL HISTOPATHOLOGICAL LESIONS BE A GUIDE TO MATERNAL AND NEONATAL OUTCOMES IN PATIENTS WITH PREECLAMPSIA?

Objective: The aim of this study was to investigate the predictive role of massive perivillous fibrinoid deposition (MPFD), syncytial knots, and accompanying histopathological features of placentas of preeclampsia (PE) on maternal and neonatal outcomes. Matherials and methods: A retrospective clinicopathological study was conducted in a tertiary unit. In the study, 51 pregnant women admitted with PD and 55 normotensive healthy pregnant women matched for age and gestational age were compared. Information regarding clinical characteristics, neonatal findings, and placental properties such as syncytial knots, vascular structure density, placental area, volume, and weight) was retrieved. Results: Massive perivillous fibrinoid deposition, syncytial knots and decreased vessels in terminal villi were significantly frequent in the PE group compared to the controls. However, these histopathological findings were not associated with clinical and neonatal outcomes. Conclusions: Syncytial knot and perivillous fibrin deposition are significant microscopic findings of preeclampsia. However, the presence and amount of fibrin deposition were not correlated with perinatal outcome.