Morfofunctional and Molecular Changes in Placenta and Peripheral Blood in Preeclampsia and Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM) and preeclampsia (PE) are common pregnancy complications with similar risk factors. Although GDM is associated with PE, the exact mechanism underlying the association is unclear. The objective of this work was to study the morphofunctional and molecular changes in the placenta and peripheral blood in PE and GDM. Local and systemic changes in the production of several placental proteins were assessed along with markers of inflammation and metabolic disorders. Expression of placental lactogen, trophoblastic ß1-glycoprotein, placental alpha-1-microglobulin, and proteinase 3 in villi was found to change in complicated pregnancy groups. Similarity of underlying pathogenic mechanisms was demonstrated for PE and GDM.

[The role of mast cells in physiological and complicated pregnancy]. / Rol' tuchnykh kletok pri fiziologicheskoi i oslozhnennoi beremennosti.

Mast cells (MCs) are highly differentiated and multifunctional immune cells. The importance of TC has been established not only as mediators of allergic reactions, but also for the development of an immune response, the occurrence of certain autoimmune diseases, tissue homeostasis, the formation of immunotolerance and metastasis of malignant tumors. MCs are present in the endometrium of women in various value depending on age, the phase of the menstrual cycle, the presence of pregnancy. Out of pregnancy, MCs are involved in the cyclic transformation of the uterine mucosa. At the onset of pregnancy, MCs stimulate the process of remodeling of the spiral arteries, the production of leukemia-inhibiting factor (LIF), which is the main implantation factor, and contribute to the formation of an immunotolerant state of the mother in relation to the fetus. Obstetric complications are accompanied by a variable content of MCs, which is associated with different genesis of diseases. A low amount of MCs is associated with impaired implantation and the development of early preeclampsia, an increased content of MCs is observed in the presence of a pathological inflammatory reaction that accompanies late preeclampsia. This review is devoted to the significance of MCs and their mediators in the physiological course of pregnancy, as well as their participation in the pathogenetic mechanisms of pregnancy complications.

[Clinical and morphological characteristics of the atypical placentation spectrum in the uterus]. / Kliniko-morfologicheskaya kharakteristika spektra atipichnoi platsentatsii v matke.

BACKGROUND: The concern of the global community of gynecologists and obstetricians (FIGO) regarding the increase in the number of caesarean sections has resulted in the creation of a new classification, Placenta Accreta Spectrum (PAS), which presents degrees of villus invasion into the uterine wall. OBJECTIVE: Compare the main types of atypical placentation (AP) with the stages of PAS, to supplement and unify the clinical and morphological criteria AP. MATERIAL AND METHODS: Surgical material was examined from 73 women after metroplasty (n=61) and hysterectomies (n=12) from the regions of Russia, Moscow and the Moscow region for ingrown villi and from 10 women with a typical placenta location during the first cesarean section. A targeted cutting of material from the uteroplacental region was used, at least 10-12 pieces, with further H&E and Mallory staining. RESULTS: In the classification of AP, the terms «placenta accreta¼, «increta¼, «percreta¼ should be retained. It is necessary to single out pl. previa as a separate type. Attention is focused on the need to assess the depth of villi invasion accompanied by a layer of fibrinoid, the volume of scar tissue and the degree of disorganization of the myometrial bundles, the state of the vessels in the serous membrane. A new type of AP has been proposed - a sharp thinning of the lower segment of the uterus, due to the scar failure and the pressure of the growing amniotic sac, leading to atrophy and necrosis of the myometrium. CONCLUSION: An integrated approach should be used to classify atypical placentation, taking into account not only the depth of villus invasion, but also anatomical and pathogenic factors in order to develop targeted methods of surgical treatment.

Regulation of the Placental Renin-Angiotensin-Aldosterone System in Early- and Late-Onset Preeclampsia.

Preeclampsia (PE) is one of the most dangerous complications of pregnancy, characterized by hypertension, proteinuria, and symptoms of multiple organ failure, which are detected de novo after 20 weeks of pregnancy. The renin-angiotensin-aldosterone system (RAAS) is one of the first to recognize pregnancy and is an important regulator of blood pressure. The placenta has its own RAAS, the role of which in the development of PE is not fully understood. In this work, for the first time, we characterized the expression of RAAS components and miRNAs controlling it in the placenta at various times of PE manifestation. The data obtained will allow the development of a new strategy in the future for the search for therapeutic agents for patients suffering from PE and cardiovascular diseases.

Telocytes in the Myocardium of Children with Congenital Heart Disease Tetralogy of Fallot.

Telocytes, a new type of interstitial stem cells with long thin processes that form a three-dimensional network around cardiomyocytes, vessels, and nerve fibers were described in the myocardium of children with tetralogy of Fallot. Two types of morphologically different telocytes, spindle-shaped and rounded, were identified. Contacts of telocytes with stem cells and interstitial macrophages were found. Telocytes were more common in the immature myocardium, where the assembly of myofibrils in cardiomyocytes was not completed and small Ki-67+ cardiomyocyte progenitor cells were present. Telocytes expressed immunohistochemical markers CD117, vimentin, CD34, and CD44. Localization and ultrastructural characteristics of telocytes suggested their participation in stem cell differentiation, coordination of neoangiogenesis, and paracrine regulation of all components of the interstitium.

Enhanced Expression of TLR8 in Placental Tissue in Preeclampsia.

The expression of TLR8 in the placental tissue was studied in specimens from women of reproductive age with early- and late-onset preeclampsia (12 and 8 patients, respectively). The reference groups included 15 women: 10 with uneventful full-term pregnancy and 5 with preterm operative delivery on gestation weeks 28-33. The expression of TLR8 in placental structures was maximum in early-onset preeclampsia (p<0.01) characterized by the gravest clinical course, while the expression of TLR8 in late-onset preeclampsia was comparable with that in full-term pregnancy. This significant increase of TLR8 expression in placental tissue seemed to reflect activation of the key proinflammatory factors of congenital immunity and induction of the systemic inflammatory response. Manifest differences in the expression of TLR8 in late- and early-onset preeclampsia confirmed the hypothesis on different variants of this condition.

Content of Free Fetal DNA in Maternal Blood and Expression of DNA Recognition Receptors ZBP-1 in Placental Tissue in Preeclampsia and Preterm Labor.

We evaluated the content of cell-free fetal DNA in maternal blood and expression of ZBP-1 receptors in the placental tissue of women with uncomplicated pregnancy, preeclampsia, and preterm labor. The study included 16 women with preeclampsia (early and late-onset preeclampsia, 8 cases each), 16 women with preterm labor, and 21 women with uncomplicated pregnancy. The concentration of cell-free fetal DNA was measured by PCR by detecting hypermethylated region of the RASSF1A gene. Immunohistochemistry was performed on paraffin-embedded sections of the placenta samples using primary polyclonal antibodies to ZBP-1. Significant increase in the level of cell-free fetal DNA was found in women with preeclampsia (both early and late-onset form) in comparison with uncomplicated pregnancy. The concentration of cell-free fetal DNA in preterm labor group did not differ from the control group; however, it was significantly lower than in early-onset preeclampsia, but not late preeclampsia. Immunohistochemical study showed higher expression of ZBP-1 in the villus syncytiotrophoblast in early-onset preeclampsia in comparison with that in preterm labor group (p=0.006). Fragments of damaged placental cells, predominantly trophoblast, enter maternal circulation and are the source of cell-free fetal DNA and a potential ligand for ZBP-1, which leads to further cell damage and the formation of a vicious circle. The increase in the content of cell-free fetal DNA in maternal blood and ZBP-1 expression in the syncytiotrophoblast in preeclampsia are interrelated processes reflecting impaired morphofunctional state of the placenta.

Peculiarities of RIG-1 Expression in Placental Villi in Preeclampsia.

The expression of RIG-1 in placenta samples was assessed in women of reproductive age with early- and late-onset preeclampsia and cesarean delivery at 27-39 weeks of gestation. The highest expression of RIG-1 was found in the syncytiotrophoblast of placental villi in the group with uncomplicated full-term pregnancy (normal); RIG-1 expression in groups with early- and late-onset preeclampsia was significantly (p<0.01) lower. In decidual cells, RIG-1 expression was also maximum in normal pregnancy and significantly (p<0.01) lower in lateonset preeclampsia. In the endothelium of villous capillaries, the maximum expression was observed in normal full-term pregnancy and in late-onset preeclampsia, while in early-onset preeclampsia this parameter was significantly (p<0.01) lower. It can be assumed that different variants of preeclampsia are mediated by similar pathogenetic mechanisms, including those related to immature molecular profile of the trophoblast and decidual cells, probably due to impaired stem cell activity in the placenta determining higher vulnerability and reduced regeneration capacity of the placental tissue. This is due to the fact that RIG-1 is one of the important signaling molecules that promote activation of stem cell and tissue regeneration.

Peculiarities of the Expression of TLR4 and Inhibitor of TLR-Cascade Tollip in the Placenta in Earlyand Late-Onset Preeclampsia.

We studied the peculiarities of the expression of TLR4 and its inhibitor Tollip in placentas obtained from women aged 23-40 years with early- and late-onset preeclampsia. Histological examination of placental tissue (hematoxylin and eosin staining) and immunohistochemical analysis with primary monoclonal antibodies to TLR4 and Tollip were performed on serial paraffin sections. It was found that the expression of TLR4 increased with increasing gestation term in both the syncytiotrophoblast and vascular endothelium of the placental villi (p<0.05). The expression of TLR4 in syncytiotrophoblast and in the endothelium in early preeclampsia was also significantly (p<0.00001) higher than in the reference group (preterm birth before 34 weeks gestation). In the vascular endothelium of placental villi, the expression of TLR4 in placentas from women with early-onset preeclampsia was higher than in late-onset preeclampsia (p=0.002), while Tollip was lower in early-onset preeclampsia than in the reference group. In preeclampsia, especially in early-onset preeclampsia, marked changes in the expression of TLR4 and Tollip in the placental tissue were detected; the severity of preeclampsia correlated with the degree of damage to the placental villi.

Change in OncomicroRNA Expression in the Placenta during Preeclampsia.

The expression of microRNA-17, microRNA-181a, and microRNA-519a in the villous tree in preeclampsia was analyzed using chromogenic in situ hybridization technique (CISH). It was found that in early-onset preeclampsia, the expression of microRNA-17 in the syncytiotrophoblast was higher (p<0.05) than in late preeclampsia, and the expression of microRNA-519a was higher (p<0.05) than in women with preterm birth at 26-31 weeks gestation. We revealed higher level of expression of microRNA-181a (p<0.05) in the cytoplasm of the syncytiotrophoblast of intermediate placental villi in the group with premature delivery in comparison with early preeclampsia. In full-term pregnancy, the expression of microRNA-181a in the vascular endothelium of placental villi was higher (p<0.02) than in women with premature deliveries. Analysis of the target genes associated with these microRNAs showed that damage to the trophoblast typical of preeclampsia, especially up to 34 weeks gestation, was accompanied by selective activation of genes participating in invasion and compensatory suppression of oncoprotective genes associated with the development of malignant neoplasms.

Dynamics of Renin, Angiotensin II, and Angiotensin (1-7) during Pregnancy and Predisposition to Hypertension-Associated Complications.

Changes in the levels of rennin, angiotensin II, and angiotensin (1-7) were studied during normal pregnancy. The blood was taken on gestation days 140-237 and 238-280. No significant changes in renin concentration were observed during normal pregnancy (p=0.423). The level of angiotensin II increased during normal pregnancy from 9.7±1.2 to 14.7±1.9 pg/ml (p=0.019). On the contrary, angiotensin (1-7) concentration decreased from 771.1±44.2 to 390.7±13.9 pg/ml (p<0.001). The shift in the proportion between vasoconstrictor angiotensin II and vasodilaltor angiotensin (1-7) attests to high predisposition of pregnant women to hypertension-related complications.

DAI-1 Receptor Expression in Placenta in Earlyand Late-Onset Preeclampsia.

DAI-1 receptor (DNA-dependent activator of IFN-regulatory factors; DLM-1/ZBP-1) is an innate immunity cytoplasmic receptor of the DNA-recognition receptor class of antiviral immunity. DAI-1 expression reflects the severity of the inflammatory response that plays the key role in the pathogenesis of pregnancy complications. We studied DAI-1 receptor expression in the placental villi in early- and late-onset preeclampsia. In case of early-onset preeclampsia DAI-1 staining intensity was lower (p=0.01), and in case of late preeclampsia - significantly higher (p<0.005) than in the reference groups at the corresponding gestational age. There was revealed a correlation between the decrease in DAI-1 receptor expression and the severity of disease progression.

Expression of MicroRNA-146a and MicroRNA-155 in Placental Villi in Early- and Late-Onset Preeclampsia.

We studied the expression of microRNA-146a and microRNA-155 in placental villi from 18 women (26-39 weeks of gestation) of reproductive age with early- or late-onset preeclampsia. The reference group consisted of women with physiological pregnancy and full-term gestation and with preterm birth after caesarian section on gestation week 26-31. MicroRNA-146a and microRNA-155 were detected by in situ hybridization with digoxigenin on paraffin sections. It was found that the expression of microRNA-146a in both syncytiotrophoblast of the intermediate villi and syncytial knots was lower at late-onset preeclampsia than at physiologic pregnancy of full-term period (p=0.037 and p=0.001 respectively). The expression of microRNA-155 in syncytiotrophoblast of intermediate placental villi in early-onset preeclampsia was higher than in group with preterm delivery (p=0.003). However, in syncytiotrophoblast of intermediate villi and in syncytial knots, the expression of microRNA-155 was lower at late-onset preeclampsia in comparison with full-term physiological pregnancy (p=0.005). In addition, the expression of microRNA-146a and microRNA-155 did not increase in the later terms in preeclampsia, while in the reference groups demonstrating gradual increase in the expression of these markers with increasing gestational age. Expression microRNA-146a and microRNA-155 little differed in early- and late-onset preeclampsia. These findings suggest that different variants of preeclampsia are probably characterized by common pathogenetic pathways. Damaged trophoblast cannot maintain of microRNAs synthesis at the required level, which determines the formation of a vicious circle in preeclampsia and further progression of the disease.

Ultrastructural Characteristics of Placental Telocytes.

Telocytes of placental villi were studied by electronic microscopy during physiological pregnancy. Ultrastructural features of telocytes indicating their heterogeneity and presence of at least three types of villi depending on their localization and kind were observed. All placental telocytes were characterized by small amount of organelles including mitochondria. Presence of long thin processes, which generated a branching network by contacting with each other, served as a typical feature of telocytes including telocytes of the stroma and intermediate villi. Telocytes were absent in the terminal villi.

Conditions for Collection of Placental Tissue Samples for Culturing of Multipotent Mesenchymal Stromal Cells.

Placentas from women aged 25-32 years with normal course of gestation were studied. It is essential to stick to certain methodological approaches for preparing viable multipotent mesenchymal stromal cell culture and to carry out morphological (macro and micro) evaluation of the chorionic villi, umbilical cords, and placentas. At stage I of the study, patients' histories, labor course, and examinations of the newborns should be analyzed to exclude women with genital and extragenital diseases. At stage II, it is essential to stick to special regulations and methods for collection of specimens of the cord, amnion, and placental tissue proper. Histological control of the placental structures collected for multipotent mesenchymal stromal cell culturing is obligatory.

[Morphological Features of Mesenhymal Stroma Cells of Chorionic Villi].

Background: Nowadays autologous mesenchymal placental stromal cells (MSCs) may use to treat for various diseases both of the mother and the child. Stroma of the placenta villi is appropriated origin for cell culture isolation. Aim: of the study was to evaluate the possibility for selection and use of placental tissue for mesenchymal stromal cells. Materials and methods: The present study was based on 45 placental samples of women aged 27−38 yy. who underwent surgical delivery at 36−40 weeks of gestation. 30 of these women have been enrolled in the basic group including children with congenital abnormalities (CA). The comparison group consisted of 15 patients with physiological pregnancy. We performed histological examination (with hematoxylin and eosin staining), immunohistochemical examination (with use monoclonal antibodies CD90 (1:25; Abcam, UK), СD105 (1:500; Abcam, UK), CD44 (1:25; Dako), СD73 (1:200, Abcam, UK), and electron microscopy (by microscope Philips/FEI Corporation, Eindhoven, Holland). Eclipse 80i microscope (Nikon Corporation, Japan) was used to examine the immunohistochemical reactions as a brown staining. The evaluation of the intensity of reaction was conducted by NIS-Elements Advanced Research 3.2 program (Czech Republic). Student's t-test and analysis of variance were used to compare the mean values. Differences were considered statistically significant at p<0.05. Results: Interstitial cells of the stroma of the villi with CA had fibroblastic differentiation as revealed degenerative changes of the cells. The histologic examination with hematoxylin and eosin staining revealed significant fibrosis of the stroma of the placenta villi in CA group (p<0,01). Immunohistochemical study of stem and intermediate chorionic villi revealed no significant differences in staining of CD44+, СD90+, СD73+, and CD105+ cells if compared to the control group (p>0.05). Although CD105 expression was significantly lower in the CA group (0.058±0.0049) than in the control group (0.088±0.0039) (p<0.05). However, electron microscopy detected the villi interstitial stromal cells with fibroblastic differentiation in CA group. Conclusions: Thus, it is necessary to exclude placenta with obstetrical history, somatic, and congenital pathology of the mother and the child when selecting the placental cell culture. Moreover, choosing a sample the morphological structure of the placenta should be taken into consideration. However, congenital malformations of the fetus, pathology of the mother cultivate mesenchymal stromal cells of placentas is inappropriate and should be taken advantage of the donor cells.

[Twin reversed arterial perfusion syndrome in the monochorionic diamniotic placenta with the development of an acardiac fetus]. / Sindrom obratnoi arterial'noi perfuzii v monokhorial'noi diamnioticheskoi platsente s razvitiem ploda-akardiusa.

The article describes an acardiac fetus in a patient with monochorionic diamniotic twin pregnancy with reversed arterial perfusion syndrome at 30 weeks' gestation. It gives postmortem fetal computed tomographic and pathoanatomic data. Microarray of acardiac fetal tissues revealed that there was deletion of chromosome 19 - arr [hg19] 19p13.3q11 (260,911-23,005,001) x1; size, 23 Mbp; the signal level in about 30% of fetal tissue cells containing deletion.

[Role of twin reversed arterial perfusion syndrome in the development of fetus acardius]. / Rol' sindroma obratnoi arterial'noi perfuzii bliznetsov v razvitii ploda-akardiusa.

Monochorionic multiple pregnancy has a higher risk for obstetric complications, including those due to the development of twin-to-twin transfusion syndrome and reversed arterial perfusion syndrome (TRAP sequence). The latter occurs in 0.1% of all monochorionic pregnancies. The basis for TRAP sequence is a relationship between arterial and venous anastomoses from the appropriate placental areas, causing a recipient fetus to develop at the expense of a donor fetus. The presence of abnormal anastomoses is considered to be a main cause of acardia. The prognosis for a donor fetus is also unfavorable: the mortality rates in the absence of intrauterine correction are as high as 55%.

Differences of glycocalyx composition in the structural elements of placenta in preeclampsia.

INTRODUCTION: Glycans expressed in the fetal-maternal interface were shown to exert immunomodulating effects and to mediate interactions between the cells. The aim of this study was to investigate alterations in the structure of carbohydrate chains of glycocalyx in placental tissue in pregnancies complicated with preeclampsia (PE). METHODS: A histochemical analysis of placental tissues was performed with a panel of biotinylated lectins. We analyzed placental tissues in women who had severe or moderate PE and compared them to placentas from women with normal pregnancies. RESULTS: There was decreased content of terminal residues of α(2,6)-linked sialic acid (as stained by SNA lectin) in the carbohydrate chains of glycocalyx of the endothelium of placental terminal villi in patients with moderate preeclampsia. The composition of the glycocalyx of syncytiotrophoblast in patients of this group did not differ from the control group. Amount of the glycans with terminal ß-Gal- (ECL) and α-mannosyl residues (ConA) in the syncytiotrophoblast and capillary endothelium of the placenta was significantly higher in the group with severe PE compared to the control group. The increased content of sialoglycans with α(2,6)-linked sialic acids residues were discovered in the syncytium, and the decreased content of α(2,3)-linked sialic acids residues - in the endothelium of terminal villi in preeclampsia. DISCUSSION: The most prominent alteration of the glycocalyx composition was found in the placentas of women with severe preeclampsia. It is likely that the modified glycome of syncytiotrophoblast and capillary endothelium may play an important role in pathogenesis of preeclampsia.

Specific Features of TLR4 Expression in Structural Elements of Placenta in Patients with Preeclampsia.

We studied the expression of TLR4 (Toll-like receptor 4) in the syncytiotrophoblast and vascular endothelial cells of terminal and stem placental villi in severe preeclampsia and normal pregnancy. In women of both groups, the expression of TLR4 was higher in the syncytiotrophoblast that in placental endothelial cells. In patients with severe preeclampsia, TLR4 expression in endothelial cells of terminal villi was 1.3-fold lower than in normal pregnancy. Lower TLR4 expression in the terminal villi endothelium in preeclampsia can underlie impaired recognition of damaging molecules with subsequent development of endothelial dysfunction and changes in immunological tolerance.