DigitalUM :: Browsing by Subject "Placenta"

Browsing by Subject "Placenta"

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Open Access
Caracterización histológica y expresión inmunohistoquímica de los marcadores p53 y p21 en el trofoblasto placentario en preeclampsia
(2017-01-04) Oviedo Ramírez, María Isabel; Pérez Cárceles, María Dolores; Blanco Carnero, José Eliseo; Escuela Internacional de Doctorado
Introducción La preeclampsia es una enfermedad frecuente en el embarazo, que se incluye dentro del grupo denominado trastornos hipertensivos del embarazo y constituye hasta un 10% de las gestaciones de forma global. Se ha encontrado incremento de la apoptosis en placentas de mujeres con preeclampsia en comparación con mujeres con embarazos sin esta enfermedad. Existen escasas referencias en relación a la expresión inmunohistoquímica de la p53 y la p21 en el citotrofoblasto y sincitiotrofoblasto de placentas de mujeres con preeclampsia por lo que el presente estudio contribuye al respecto, complementando con la aportación de la expresión de la apoptosis a nivel óptico evaluada con hematoxilina y eosina. Objetivos Los objetivos de este trabajo son describir los principales hallazgos histopatológicos en placentas con preeclampsia, así como describir la expresión inmunohistoquímica de los marcadores p53 y p21. Material y métodos En esta investigación estudiamos placentas procedentes de mujeres del área de salud I (Murcia/Oeste) que durante el embarazo presentaron preeclampsia. Se utilizó un grupo control de mujeres con gestaciones normales. El estudio se realizó en colaboración con el Área de Fisiología de la Facultad de Medicina de la Universidad de Murcia y la Unidad de Medicina Materno fetal del Hospital Clínico Universitario Virgen de la Arrixaca, del cual uno de los facultativos especialistas obtuvo los datos clínicos y muestras biológicas. El estudio histopatológico y la inmunohistoquímica se realizó en el Servicio de Anatomía Patológica de este Hospital. El trabajo fue de tipo observacional, transversal y prospectivo. La muestra final incluyó 53 placentas, de estás 15 correspondieron a placentas de embarazo normal (grupo control), 22 a placentas con preeclampsia pretérmino y 16 a placentas con preeclampsia a término. El estudio microscópico se realizó con tinción de hematoxilina y eosina obteniéndose varias preparaciones histológicas de cada caso. La evaluación inmunohistoquímica se realizó siguiendo la escala de Fulop con marcadores para p53 y p21. Resultados Las placentas con preeclampsia mostraron signos de arteriolopatía decidual (aterosis aguda, necrosis fibrinoide y placentación deficiente), sin evidencia de cambios de este tipo en las placentas del grupo control con embarazo normal. La arteriolopatía decidual en placentas con preeclampsia fue alta (65,8%), encontrándose además mayor número de infartos (55,3%). Por otro lado se encontraron diferencias notables entre las placentas con preeclampsia del grupo pretérmino y a término, observando mayor número de casos de arteriolopatía decidual, aterosis aguda, necrosis fibrinoide y placentación deficiente en el grupo de placentas con preeclampsia pretérmino que en el de término (72,7%, 54,6%,45,4% y 77,3%). Las placentas con preeclampsia pretérmino mostraron mayor incremento de células de hofbauer, cambios estromales y grado severo de presencia de nodos sincitiales ( >50%) siendo estadísticamente significativo. Además se encontró que las placentas con preeclampsia a término mostraron más calcificaciones que el grupo de preeclampsia pretérmino, con diferencias estadísticamente significativas (93,8%). En relación a los estudios de inmunohistoquímica realizados, se encontró que tanto la intensidad, como el grado de expresión de la p53 y p21 en el citotrofoblasto, fue mayor en las placentas con preeclampsia que en el grupo control. La inmunohistoquímica de la p53 en el sincitiotrofoblasto no mostró diferencias, que si fueron detectadas en la tinción de p21 en el sincitiotrofoblasto tanto en intensidad como en el grado de expresión. En cuanto a los dos grupos de placentas de preeclampsia, no se encontraron diferencias significativas en intensidad y grado de expresión de p53. La intensidad de la p21 tanto en el citotrofoblasto como en el sincitiotrofoblasto fue mayor en placentas con preeclampsia pretérmino con tinción marcada en 77,3% y 50% respectivamente mostrando además mayor grado de expresión para este marcador. Conclusiones El grupo de placentas con preeclampsia mostró significativamente una mayor frecuencia de aterosis aguda, necrosis fibrinoide, infartos y placentación deficiente, contrastando con el grupo de placentas sin enfermedad, en las que no se detectaron estos cambios. Las placentas con preeclampsia mostraron diferencias histopatológicas respecto a placentas no patológicas, identificándose una mayor frecuencia de arteriolopatía decidual, lo que evidencia una mayor susceptibilidad al daño vascular estructural. La intensidad y el grado de expresión inmunohistoquímica de la p53 en el citotrofoblasto fue significativamente superior en las placentas con preeclampsia que en el grupo control. La tinción inmunohistoquímica de p21 fue mayor en el citotrofoblasto en las placentas con preeclampsia, tanto en la intensidad como en el grado de expresión, que en el grupo control. También en el sincitiotrofoblasto el grado de expresión fue significativamente superior en el grupo de preeclampsia, que en el grupo sin patología. Las placentas con preeclampsia pretérmino mostraron significativamente una mayor expresión inmunohistoquímica de la p21 en el citotrofoblasto, que en los casos de preeclampsia a término. Observamos una asociación significativa entre la presencia de altas resistencias uterinas y un mayor grado de expresión inmunohistoquímica de la p21 en el citotrofoblasto, hallazgo que podría constituir una interesante línea de investigación futura. SUMMARY Introduction Preeclampsia is a common disease during pregnancy, and is included within the group of diseases known as hypertensive pregnancy disorders and accounts for 10% of overall gestations. Increased apoptosis has been found in the placentas of women with pre-eclampsia unlike women with pregnancies without this disease. There have been few references to p53 and p21 immunohistochemical expression in the cytotrofoblast and syncytiotrofoblast of the placentas of women with pre-eclampsia so that the present study intends to contribute to this area of research, complemented by an analysis of apoptosis at an optical level assessed using hematoxylin and eosin. Objectives The objectives of this study are to describe the main histopathological findings in placental samples of patients with preeclampsia, and to report on the detection of p53 and p21 proteins by immunohistochemistry. Material and methods In our present research we have studied placentas from women in our Healthcare Area I (Murcia/West) who presented with preeclampsia during pregnancy. A group of women with normal gestation was used as the control. The study was carried out in collaboration with the Physiology Department of the Faculty of Medicine at the University of Murcia and the Maternal-Fetal Medicine Unit at the Virgen de la Arrixaca Clinical Hospital, where one of the specialist physicians obtained the clinical data and biological samples. The histopathological study and immunohistochemistry were carried out in the Pathological Anatomy Service at the same hospital. It was a prospective, observational and cross-sectional study. The final sample included 53 placentas, of which 15 were placentas from normal pregnancies (control group), 22 placentas with preeclampsia from preterm pregnancies and 16 placentas with preeclampsia from term pregnancies. The microscopic analysis was carried out using hematoxylin and eosin staining obtaining several histological preparations from each case. The immunohistopathological assessment was carried out following Fulop’s scale with markers for p53 and p21. Results The placentas with preeclampsia showed signs of decidual arteriolopathy (acute atherosis, fibrinoid necrosis and deficient placentation), while there was no evidence of these types of changes in the placentas of the control group (with a normal pregnancy). In placentas with preeclampsia, decidual arteriolopathy was high (65.8%), and there were also a high number of heart attacks (55.3%). In addition there were notable differences between the placentas with preeclampsia in the preterm and the term pregnancy group, with more cases of decidual arteriolopathy, acute atherosis, fibrinoid necrosis and deficient placentation being observed in the group of placentas with preeclampsia in the preterm pregnancy group than in the term group (72.7%, 54.6%,45.4% and 77.3%). The placentas with preeclampsia in the preterm group had a greater increase in Hofbauer cells, stromal changes and a severe level of syncytial nodes present ( >50%), all being statistically significant. Additionally, it was found that the placentas with preeclampsia in term pregnancies had more calcifications than in the preterm pregnancy group with a statistically significant difference (93.8%). With regard to the immunohistochemical analysis carried out, it was found that both the intensity of p53 and p21 in the cytotrophoblast and the grade of expression of these markers, was greater in the placentas with preeclampsia than in the control group. Immunochemical evidence of p53 protein in the syncytiotrophoblast did not show any differences between the groups, detected using p21 staining in the syncytiotrophoblast to analyze the intensity and grade of expression. With regard to the two groups of placentas with preeclampsia, no significant differences were found in p53 protein intensity and expression. The intensity of p21 expression in both cyto- and syncytiotrophoblast was greater in the placentas with preeclampsia of preterm pregnancies using marker staining in 77.3% and 50% respectively, also revealing a higher level of expression for this marker. Conclusions The group of placentas with preeclampsia showed a significantly higher frequency of acute atherosis, fibrinoid necrosis, heart attacks and deficient placentation, contrasting with the group of placentas with no disease, in which no changes were detected. The placentas with pre-eclampsia showed histopathological differences compared to non-diseased placentas, with a higher frequency of decidual atheriolopathy being identified, a finding which reveals greater susceptibility to structural vascular damage. The intensity and grade of p53 immunohistochemical expression in cytotrophoblast was significantly higher in the placentas with preeclampsia than in the control group. Immunohistochemical staining with p21 had a greater intensity and grade of expression in the cytotrophoblast in the placentas with preeclampsia, than in the control group. It was also found that in the syncytiotrophoblast the grade of expression was significantly greater in the preeclampsia group, than in the group without any disease. The placentas with preeclampsia from preterm pregnancies had a significantly higher immunohistochemical expression of p21 in the cytotrophoblast, than in cases of pre-eclampsia in term pregnancies. We have found a significant association between the presence of high uterine artery resistance and a higher immunohistochemical expression of p21 protein in the cytotrophoblast, a finding that could provide an interesting line of research in the future.
Open Access
CCM2 and CCM3 proteins contribute to vasculogenesis and angiogenesis in human placenta
(Murcia : F. Hernández, 2009) Tanriover, Gamze; Seval, Yasemin; Sati, Leyla; Gunel, Murat; Demir, Necdet
Placenta as an ideal model to study angiogenic mechanisms have been established in previous studies. There are two processes, vasculogenesis and angiogenesis, involved in blood vessel formation during placental development. Therefore, blood vessel formation is a crucial issue that might cause vascular malformations. One of the vascular malformations is cerebral cavernous malformation (CCM) in the central nervous system, consisting of endothelium-lined vascular channels without intervening normal brain parenchyma. Three CCM loci have been mapped as Ccm1, Ccm2, Ccm3 genes in CCM. In order to investigate whether CCM proteins participate in blood vessel formation, we report here the expression patterns of CCM2 and CCM3 in developing and term human placenta by means of immunohistochemistry and Western blot analysis. CCM2 and CCM3 were obviously detected in the vascular endothelium during early pregnancy. Moreover, vascular endothelium of stem villi revealed a moderate immunostaining for CCM2 and, to a lesser extent, in the endothelium of mature intermediate villi in term placenta. Interestingly, CCM3 immunostaining was weakly localized in the endothelium of mature intermediate villi and showed lesser expression going toward stem villi in term placenta. The expression patterns of the proteins were clearly identified in the vascular endothelium of human placenta, suggesting that they might play roles during angiogenesis and vasculogenesis. Furthermore, with this study, CCM2 and CCM3 have been described for the first time in the human placenta.
Open Access
Cell cycle inhibitor p57 expression in normal and diabetic rat placentas during some stages of pregnancy
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2012) Acar, Nuray; Turkay Korgun, Emin; Ustunel, Ismail
Placentomegaly, an abnormal increase in the size of the placenta, is commonly seen in human diabetic pregnancies and diabetic animal experimental models. Proper placental development depends on the proliferation and differentiation of trophoblasts. However, our knowledge about the mitotic regulators that play key roles in synchronizing these events is limited. p57 is a cyclin-dependent kinase (CDK) inhibitor acting in the G1/S transition of the cell cycle. There is no data regarding p57 expression in either rat or human diabetic placentas. The purpose of this study was to investigate p57 expression in control and diabetic rat placentas at different stages of pregnancy. Diabetes was induced by streptozotocin on the first day of pregnancy, and placentas were taken on days 11, 13, 17, and 21 of pregnancy. Our results showed that on day 11, p57 immunostaining intensity was stronger in control group placentas compared to the diabetic group. On day 13, p57 immunostaining intensity increased in both groups, but increased more in the diabetic group. On day 17, p57 immunostaining intensity decreased in both the control and diabetic groups compared to day 13, yet the intensity remained higher in control placentas compared to diabetic placentas. On day 21 of pregnancy, p57 immunostaining intensity increased in the control group and it decreased from the day 17 level in the diabetic group. Western blot results showed consistency with immunohistochemistry results. Our study shows different expression patterns of p57 between control and diabetic rat placentas, which indicate p57 may play a role in abnormal placental formation resulting in placentomegaly arising from diabetes
Open Access
Changes in vascular extracellular matrix composition during decidual spiral arteriole remodeling in early human pregnancy
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Smith, Samantha D.; Choudhury, Ruhul H.; Matos, Patricia; Horn, James A.; Lye, Stephen J.; Dunk, Caroline E.; Aplin, John D.; Jones, Rebecca L.; Harris, Lynda K
Uterine spiral arteriole (SA) remodeling in early pregnancy involves a coordinated series of events including decidual immune cell recruitment, vascular cell disruption and loss, and colonization by placentalderived extravillous trophoblast (EVT). During this process, decidual SA are converted from narrow, muscular vessels into dilated channels lacking vasomotor control. We hypothesized that this extensive alteration in SA architecture must require significant reorganization and/or breakdown of the vascular extracellular matrix (ECM). First trimester decidua basalis (30 specimens) was immunostained to identify spiral arterioles undergoing trophoblast-independent and -dependent phases of remodeling. Serial sections were then immunostained for a panel of ECM markers, to examine changes in vascular ECM during the remodeling process. The initial stages of SA remodeling were characterized by loss of laminin, elastin, fibrillin, collagen types III, IV and VI from the basement membrane, vascular media and/or adventitia, and surrounding decidual stromal cells. Loss of ECM correlated with disruption and disorganization of vascular smooth muscle cells, and the majority of changes occurred prior to extensive colonization of the vessel wall by EVT. The final stages of SA remodeling, characterized by the arrival of EVT, were associated with the increased mural deposition of fibronectin and fibrinoid. This study provides the first detailed analysis of the spatial and temporal loss of ECM from the walls of remodeling decidual SA in early pregnancy.
Open Access
Comparative analyses on expression of galectins1-4, 7-10 and 12 in first trimester placenta, decidua and isolated trophoblast cells in vitro
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Unverdorben, Laura; Jeschke, Udo; Santoso, Laura; Hofmann, Simone; Kuhn, Christina; Arck, Petra; Hutter, Stefan
Introduction: Galectins are members of the mammalian β-galactoside-binding proteins, which recognize Galβ1-4GlcNAc sequences of several cell surface oligosaccharides. Plenty of galectins are already described in human tissue, especially in placenta. Here, gal-1-4, 7-10 and gal-12 were investigated systematically in trophoblast and decidua cells of first trimester placentas. Material and methods: Within this study, 15 first trimester placentas after induced abortion (7th-14th week of gestation) were examined with immunohistology and immunofluorescence based on a scoring system. Moreover, isolated and cultivated trophoblast cells from the first trimester were analyzed and evaluated for expression of gal-1-4, gal-7-10 and gal-12 at mRNA and protein level with real-time RTPolymerase chain Reaction/PCR (Taq-Man). Double immunofluorescence with trophoblast specific markers identified galectin expressing cells at the feto-maternal interface. Results: We could detect immunohistochemical staining of galectins 1-4, 7-10 and 12 in first trimester placenta: all examined galectins were found in the cytotrophoblast (CTB) and syncytiotrophoblast (SCT). Gal-1, -2, -3, -4, -7, -8, -9, -10 and -12 were identified in extravillous trophoblast cells (EVT) in immunohistology and immunoflourescence. The expression of gal-1, -9, - 10, and gal-12 increased after 96h incubation in vitro without stimulation at mRNA level, while gal-2, -3, -4, -7 and -8 were decreased. Discussion and conclusion: This study describes a systematic analysis of the expression of gal-1-4, gal-7-10 and gal-12 in first trimester placentas and isolated trophoblast cells. Expression levels at mRNA level and the change within 96h cultivation in vitro indicate a possible influence on syncytium building of trophoblast cell on expression of galectins. Therefore, an interaction of galectins in vitro in syncytium building is possible.
Open Access
Crim1–, a regulator of developmental organogenesis
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Iyer, Swati; Pennisi, David J.; Piper, Michael
The regulation of growth factor localization, availability and activity is critical during embryogenesis to ensure appropriate organogenesis. This process is regulated through the coordinated expression of growth factors and their cognate receptors, as well as via proteins that can bind, sequester or localize growth factors to distinct locations. One such protein is the transmembrane protein Crim1. This protein has been shown to be expressed broadly within the developing embryo, and to regulate organogenesis within the eye, kidney and placenta. Mechanistically, Crim1 has been revealed to mediate organogenesis via its interaction with growth factors including TGFβs, BMPs, VEGFs and PDFGs. More recently, Crim1 has been shown to influence cardiac development, providing further insights into the function of this protein. This review will provide an overview of the role of Crim1 in organogenesis, largely focusing on how this protein regulates growth factor signaling in the nascent heart. Moreover, we will address the challenges ahead relating to further elucidating how Crim1 functions during development.
Open Access
Differential microscopic finding and glucose transporter 3 expression in terminal chorionic villi among birth weight-discordant twin placentas
(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2015) Lee, Mina; Choi, Song-Yi; Kang, Byung-Hun; Yoo, Heon-Jong; Song, Soo-Youn; Seong, In-Ock; Suh, Kwang-Sun; Kim, Kyung-Hee
Objective: To evaluate differences in microscopic findings and glucose transporter 3 (GLUT3) expression in terminal chorionic villi (TV) among birth weight-discordant twin (BWDT) placentas compared with the birth weight-concordant twin (BWCT) placentas. Methods: We retrospectively studied a cohort of 26 BWDT, 10 BWCT, 10 pre-eclampsia singleton and 10 normal singleton pregnancies. Placentas were scored for the percentage of TV, the percentage of TV with syncytial knots, the presence of capillary branching patterns of TV, the capillary to terminal villous ratios, the membranous expression of GLUT3 and the nuclear expression of HIF-1α in trophoblasts and capillary endothelial cells of TV using immunohistochemistry. The clinical characteristics and microscopic findings were analyzed and compared. Results: BWDT placentas exhibited differential percentages of TV, percentages of TV with syncytial knots, capillary to terminal villous ratios, expression of HIF-1α in capillary endothelial cells and expression of GLUT3 in trophoblasts and capillary endothelial cells of TV among each twin pair compared with BWCT placentas (P=0.003, P=0.022, P=0.037, P=0.007, P=0.046 and P=0.002, respectively). Pre-eclampsia singleton placentas exhibited higher GLUT3 expression in trophoblasts, higher HIF-1α expression in capillary endothelial cells of TV and high capillary to terminal villous ratios compared with normal singleton placentas (P=0.001, P<0.001 and P=0.001, respectively). Conclusions: We observed a strong relationship between characteristics of adaptive change to hypoxia (GLUT3 expression, TV and syncytial knotting and higher capillary to terminal villous ratios) and BWDT pregnancy but not BWCT pregnancy.
Open Access
Distal villous lesions are clinically more relevant than proximal large muscular vessel lesions of placental fetal vascular malperfusion
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2022) Stanek, Jerzy
Background. Fetal vascular malperfusion (FVM) can be diagnosed on placental examination based on histology of distal placental villi and large muscular placental vessels. While histology of both those placental compartments can be low grade or high grade, it is not known if these are clinically equivalent. This retrospective study aimed to compare the impact of placental distal villous and large vessel FVM lesions on clinical and placental phenotypes. Methods. Clinical and placental phenotypes of 479 consecutive ≥20 weeks of gestation at delivery cases of placental FVM were analyzed among 3 groups: Group 1: 86 cases with distal FVM (clusters of sclerotic distal villi and/or those with stromal vascular karyorrhexis and/or mineralization, and/or endothelial fragmentation by CD34 immunostain) without large vessel lesions; Group 2: 186 cases with large vessel lesions (fetal vascular ectasia, vascular thrombi, stem vessel obliteration, intramural fibrin deposition) without distal villous lesions; and Group 3: 207 cases showing both distal villous lesions and large fetal vessel lesions. Results. Statistically significant differences (Bonferroni correction) were observed in: average gestational age at delivery 31, 35, 34 weeks, fetal growth restriction 24, 9, 25%, average placental weight 318, 413, 366 g, postuterine pattern of chronic hypoxic placental injury 12, 2, 6%, luminal vascular abnormalities in stem vessels 16, 3, 11%, and high grade FVM 33, 16, 39%, among Groups 1-3, respectively. Conclusion. Because of longer time needed for its development, distal FVM portends poorer prognosis for the fetus than large vessel FVM
Open Access
Estudio del papel del polimorfismo Val108/158 Met del gen humano de la catecol-O-metiltransferasa y su producto, el 2-metoxiestradiol, en el origen y fisiopatología de la preeclampsia
(2015-07-02) Pertegal Ruiz, Miriam; Hernández García, Isabel; Fenoy Palacios, Francisco José; Facultad de Medicina
Los objetivos de esta tesis fueron por un lado evaluar la asociación entre el polimorfismo funcional Val108/158Met de la catecol-O-metiltransferasa (COMT) fetal y materno y el riesgo de desarrollar preeclampsia (PE), examinando la influencia de dicho polimorfismo en la expresión, la actividad enzimática placentaria de la COMT y en los niveles maternos de su producto, el 2-metoxiestradiol (2ME), en gestantes control y preeclámpticas. Por otra lado, analizamos si los niveles del 2ME están relacionados con diferentes índices de gravedad clínica y biomarcadores de la enfermedad. Llevamos a cabo un estudio caso-control observacional, analítico y prospectivo en 126 gestantes (53 preeclámpticas y 73 gestaciones normales) reclutadas en el Hospital Clínico Universitario Virgen de la Arrixaca de Murcia entre enero de 2010 y enero de 2012. Se extrajeron muestras de sangre de las gestantes antes del parto y, una vez producido el mismo, se tomó sangre del cordón umbilical (fetal) y fragmentos de tejido placentario. Las muestras sanguíneas se utilizaron para realizar los estudios de genotipado, y para medir los niveles maternos de 2ME, y de diferentes biomarcadores relacionados con la PE (homocisteína, sFlt1, PlGF). Por otra parte, en el tejido placentario se realizaron los análisis de expresión y actividad enzimática de la COMT. Finalmente, obtuvimos datos clínicos y analíticos de la historia clínica de las pacientes. Los principales resultados de este trabajo muestran que el genotipo met-met fue el doble de frecuente en la sangre fetal de gestantes preeclámpticas que en las controles. La odds ratio ajustada (ORajt) para el riesgo de PE en dichas mujeres fue de 3.22 [intervalo de confianza (IC) 95%: 1.01, 10.28]. Los análisis del polimorfismo materno de la COMT no revelaron un aumento significativo del riesgo de PE, y tampoco añadió nada el estudio de los haplotipos de la COMT. Al analizar la actividad enzimática in vitro, ésta se vio determinada por el polimorfismo fetal Val108/158Met, siendo menor para el genotipo met-met que para el val-met y el val-val tanto en población total como en la preeclámptica. La actividad de la COMT in vitro no difirió entre controles y preeclámpticas dentro de cada genotipo. En cuanto a la expresión de la COMT soluble, ésta fue menor para el genotipo met-met en ambas subpoblaciones y dicha expresión se correlacionó inversamente con los valores de actividad in vitro, pudiendo por tanto estar determinada dicha actividad por la cantidad de proteína. La expresión placentaria de la COMT soluble no difirió entre controles y preeclámpticas dentro de cada genotipo. En lo que respecta a la actividad in vivo, no encontramos ninguna asociación entre el polimorfismo estudiado y los niveles de 2ME en plasma materno; no obstante los valores de 2ME sí estuvieron significativamente disminuidos en las gestantes con PE frente a las controles (1818.41±189.25 pg/ml vs 2906.43±200.69 pg/ml). Además, las gestantes con PE presentaron unos niveles plasmáticos mayores de homocisteína, condicionando éstos un mayor riesgo de PE, y correlacionándose inversamente con los valores de 2ME, lo que podría contribuir a la menor actividad observada de la COMT in vivo en PE. Por otra parte, los valores plasmáticos de 2ME se correlacionaron con parámetros clínicos y analíticos de gravedad del síndrome como la mayor tensión arterial sistólica pico, la necesidad de una mayor agresividad terapéutica antihipertensiva, el menor peso neonatal o la precocidad de la PE. Finalmente, los niveles maternos de 2ME también se correlacionaron, tanto en la población total como en la preeclámptica, con conocidos factores de desequilibrio angiogénico en PE como el sFlt1 o el PlGF, sugiriendo que el 2ME podría ser un factor clave en el mantenimiento del equilibrio angiogénico necesario para el correcto desarrollo de la gestación. STUDY OF THE ROLE OF VAL108/158MET POLYMORPHISM OF CATECHOL-O-METHYLTRANSFERASE HUMAN GENE AND ITS PRODUCT, 2-METHOXYESTRADIOL, ON THE ORIGIN AND PATHOPHYSIOLOGY OF PREECLAMPSIA The objectives of this thesis were on one hand to assess the association between fetal and maternal catechol-O-methyltransferase (COMT) Val108/158Met functional polymorphism and the risk of developing preeclampsia (PE), examining the influence of this polymorphism in expression and activity of placental COMT enzyme and in maternal plasma levels of its product, 2-methoxyestradiol (2ME), in both control and preeclamptic pregnant women. On the other hand, we analyze whether 2ME levels are associated with different clinical severity index and biomarkers of PE. We conducted an observational, analytical and prospective case-control study in 126 pregnant women (53 preeclamptic and 73 normal pregnancies) recruited at the Hospital Clínico Universitario Virgen de la Arrixaca in Murcia from January 2010 to January 2012. Blood samples of pregnant women were taken before delivery and, immediately after, cord blood (fetal) and placental tissue fragments were taken. Blood samples were used for genotyping studies, and to measure maternal levels of 2ME, and different biomarkers related to PE (homocysteine, sFlt1, PlGF). Moreover, we made the analysis of expression and enzymatic activity in vitro of COMT in placental tissue. Finally, we obtained clinical and laboratory data from the clinical history of patients. The main results of this work show that the genotype met-met was twice as frequent in fetal blood from patients with PE as in controls. The adjusted odds ratio (ORajt) for the risk of PE in these women was 3.22 [95% confidence interval (CI): 1.01 -10.28]. Analyses of maternal COMT polymorphism did no reveal a significant increased risk of PE, and either added nothing COMT haplotypes analysis. When analyzing the enzyme activity in vitro, it was determined by fetal Val108/158Met polymorphism, being lower for met-met genotype than for val-met and val-val both the total population and preeclamptic. The COMT activity in vitro did not differ between control and preeclamptic pregnant women within each genotype. The expression of soluble COMT was lower for the met-met genotype in both subpopulations and this expression was inversely correlated with in vitro activity values, then such activity may be determined by the amount of protein. Placental expression of soluble COMT not differ between controls and preeclamptic within each genotype In respect to in vivo activity, we found no association between the polymorphism studied and levels of 2ME in maternal plasma, however those values were significantly decreased in women with PE compared to control (1818.41±189.25 pg/ml versus 2906.43±200.69 pg/ml). In addition, there were a higher plasma homocysteine levels in preeclamptic women conditioning them at increased risk of PE, and correlated inversely with the values of 2ME in total population, which could contribute to the lower COMT activity in vivo observed in PE. On the other hand, plasmatic 2ME values were significantly correlated with different clinical and laboratory parameters of severity of the syndrome such as higher peak systolic blood pressure, the need for more aggressive antihypertensive therapeutic, lower birth weight or early onset of PE. Finally, maternal 2ME levels were also correlated (in patients with PE and in total population) with known angiogenic imbalance factors present in PE as sFlt1 or PlGF, suggesting that 2ME could be a key factor in maintaining the balance angiogenic necessary for the proper development of gestation.
Open Access
Expression of the retinoblastoma-related p107 and Rb2/p130 genes in human placenta: an immunohistochemicaI study
(Murcia : F. Hernández, 2001) Cavallotti, I.; De Luca, L.; D'Aponte, A.; De Falco, M.; Acanfora, F.; Visciano, M.L.; Gualdiero, L.; De Luca, B.; Baldi, A.; De Luca, A.
It has been proposed that tumor suppressor genes may have a role in the mechanisms of proliferation and differentiation during human placental development. The Retinoblastoma gene family is a well known family of tumor suppressor genes. Many studies have pointed out a role of this family not only in cell cycle progression, but also during development and differentiation. On the light of these observations we have investigated the immunohistochemical expression pattern of the Retinoblastoma family members, p107 and Rb2/p130 in human placenta samples in first trimester and full-term placental sections. p107 and pRb2lp130 showed the most abundant expression levels during the first trimester of gestation and progressively declined to being barely detectable in the placenta by late gestation. These results indicate that the expression of the above genes is modulated during placental development and suggest a mechanism for controlling trophoblast proliferation.
Open Access
Expression of VEGF-C and activation of its receptors VEGFR-2 and VEGFR-3 in trophoblast
(Murcia : F. Hernández, 2001) Dunk, C.; Ahmed, A.
Placental villous development requires the co-ordinated action of angiogenic factors on both endothelial and trophoblast cells. Like vascular endothelial growth factor (VEGF), VEGF-C increases vascular permeability, stimulates endothelial cell proliferation and migration. In the present study, we investigated the expression of VEGF-C and its receptors VEGFR-3 and VEGFR-2 in normal and intrauterine growth-restricted (IUGR) placenta. Immunolocalisation studies showed that like VEGF and VEGFR-1, VEGF-C, VEGFR-3 and VEGFR-2 co-localised to the syncytiotrophoblast, to cells in the maternal decidua, as well as to the endothelium of the large placental blood vessels. Western blot analysis demonstrated a significant decrease in placental VEGF-C and VEGFR-3 protein expression in severe IUGR as compared to gestationallymatched third trimester pregnancies. Conditioned medium from VEGF-C producing pancreatic carcinoma (Suit-2) and endometrial epithelial (Hec-1B) cell lines caused an increased association of the phosphorylated extracellular signal regulated kinase (ERK) in VEGFR-3 immunoprecipitates from spontaneously transformed first trimester trophoblast cells. VEGF121 caused dosedependant phosphorylation of VEGFR-2 in trophoblast cells as well as stimulating DNA svnthesis. In addition. premixing VEGFl with Yheparin sÚlphate proteoglycan wtentiated trooho8ast oroliferation and the association Lf p h ~ s p hw~ith- t i~e V~E~GF R-2 receptor. VEGF165- mediated DNA synthesis was inhibited by anti-VEGFR- 2 neutralising antibody. The results demonstrate functional VEGFR-2 and VEGFR-3 receptors on trophoblast and suggest that the decreased expression of VEGF-C and VEGFR-3 may contribute to the abnormai villous development observed in IUGR placenta.
Open Access
Expression pattern alterations of the serine protease HtrA1 in normal human placental tissues and in gestational trophoblastic diseases
(Murcia : F. Hernández, 2009) Marzioni, Daniela; Quaranta, Alexia; Lorenzi, Teresa; Morroni, Manrico; Crescimanno, Caterina; De Nictolis, Michele; Toti, Paolo; Muzzonigro, Giovanni; Baldi, A.; De Luca, Antonio; Castellucci, Mario
HtrA1 is a secreted protein which behaves as a molecular chaperone at low temperatures and as a serine protease at high temperatures. When the placenta escapes the normal growth control mechanisms, which are present during normal pregnancy, it may develop trophoblastic diseases, such as hydatidiform mole and choriocarcinoma. The aim of the study is to investigate the expression of HtrA1 in these gestational trophoblastic diseases and evaluate whether different HtrA1 expression might be associated with increasingly severe forms of disease. We used immunohistochemistry to assess the expression of HtrA1 in normal human placenta, hydatidiform mole (partial and complete) and choriocarcinoma. In addition to that we used the western blotting technique to quantify HtrA1 immunoreaction in normal human placentas. The most striking finding of our investigation is the decrease in immunostaining of this protease with increasing severity of gestational trophoblastic disease. For instance, in partial and complete moles HtrA1 is weakly expressed in the trophoblast. Moreover, absence of immunoreaction for HtrA1 is observable in the choriocarcinoma cells. In conclusion, we suggest that HtrA1 may play an important role in the pathogenesis and progression of hydatidiform moles and choriocarcinomas, and that HtrA1 may play an important role during the normal development of the placenta, as well as in trophoblastic diseases.
Open Access
Glypican-3 (GPC3) expression in human placenta: localization to the differentiated syncytiotrophoblast
(Murcia : F. Hernández, 2001) Khan, S.; Blackburn, M.; Mao, D.L.; Huber, R.; Schlessinger, D.; Fant, M.
The expression of glypican-3 (GPC3), a heparan-sulfate proteoglycan associated with the Simpson-Golabi-Behmel fetal overgrowth syndrome, was studied in normal human placental tissue and cell lines derived from human placentae. Cytotrophoblasts derived from term placentae expressed GPC3 mRNA at low levels in culture. GPC3 mRNA expression increased markedly during trophoblast differentiation. By contrast, fibroblast cell lines derived from normal placentae did not express GPC3 in culture. Similarly, choriocarcinoma cell lines derived from human placentae (BeWo, JAR, and JEG) failed to express GPC3 mRNA. In situ hybridization confirmed the localization of GPC3 mRNA to the syncytiotrophoblast. Furthermore, immunohistochemical staining of paraffin imbedded placental tissue demonstrated intense staining of the syncytiotrophoblast cell layer and less intense staining of cytotrophoblasts. No staining of mesenchymal elements was noted. These data confirm the presence of GPC3 in human placenta and suggest it is expressed by the differentiated syncytiotrophoblast at term
Open Access
Immunoexpression of the hepatocyte growth factor (HGF), HGF-receptor (c-met) and STAT3 on placental tissues from malformed fetuses
(Murcia : F. Hernández, 2002) Trovato, M.; Grosso, Maddalena; Vitarelli, E.; Benvenga, S.; Trimarchi, F.; Barresi, G.
To characterize the possible relationship between the expression of the HGF/HGF-R system with transcription factor STAT3, responsible for morphogenetic response of HGF stimulation, and the embryonic development alterations, we investigated, by immunohistochemistry, the expression of HGF, c-met and STAT3 in 9 placentas from malformed fetuses and 9 control placentas from non-malformed fetuses. The major and distinct patterns of expression characterizing the placentas from malformed fetuses were a higher percentage mean of stromal cells stained for HGF, c-met and STAT3 antibodies (60%, 66% and 54%, respectively on fibroblast cells and 44%, 57% and 42%, respectively on myofibroblast cells) and a lower percentage mean of cytotrophoblast cells stained for the same antibodies (2%, 2% and 1%, respectively), than in control placentas. In fact, in this latter group, the stromal fibroblast cells were stained in a percentage mean of 27%, 22% and 7%, respectively; the stromal myofibroblast cells in a percentage mean of 5%, 6% and 2%, respectively and the cytotrophoblast cells in a percentage mean of 25%, 34% and 18%, respectively. The expression of each antibody on stromal cells in both groups suggests an alternative role of the HGF/HGF-R system activating the via STAT3 transdution and operating on placental tissues, overall in organogenesis alteration conditions. This immunohistochemical approach could be used in the diagnostic practice of pathologists on chorionic villi biopsy when genetic alterations are absent and ultrasound aspects are doubtful for malformations.
Open Access
Impact of COVID-19 disease on placental histopathology. PLAXAVID study
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2024) Montáns Araújo, J.; Suy Franch, A.; García Ruiz, I.; Maíz, N.; Garcia Aguilar, E.; Hidalgo Bermejo, F.J.
Background. The impact of COVID-19 on pregnancy has been analyzed suggesting an increased risk of placental lesions that might lead to maternal and neonatal complications. However, the current published evidence is not conclusive because contradictory results. Methods. PLAXAVID is an observational, retrospective, histopathological, single-center study that aimed to evaluate the prevalence of vascular and inflammatory lesions in placental and umbilical cord samples of one hundred women infected by SARS-CoV2 during pregnancy. Results. The histopathological analysis showed that in most of the placentas (77.8%) there were signs of maternal vascular malperfusion (MVM; primary endpoint). The most common MVM features were an accelerated villous maturation (37.4%), central villous infarcts (33.3%), and villous agglutination (46.5%). Fetal vascular malperfusion (FVM) was identified in 57.6% of samples, and the most frequent features were hyalinized avascular villi (38.4%), fetal vascular thrombi (20.2%) and umbilical cord at risk of partial obstruction (14.1%). Acute and chronic inflammatory pathology were noticed in 22.2% and 49.5% of placentas, respectively. No significant correlations were found between MVM presence and the time, duration, and severity of infection, nor with the duration of pregnancy. However, in critically ill patients, the pregnancy duration (p=0.008), newborn weight (p=0.003), and APGAR test scores (p<0.001) were significantly lower. The same trend was observed considering the presence of infection at the time of delivery and in preterm births. Conclusion. A very high percentage of placentas with vascular and/or inflammatory lesions was found in the analyzed cohort. Therefore, PLAXAVID study results supported that COVID-19 should be considered a risk factor during gestation and requires close monitoring of pregnancy.
Open Access
Keratins in the human trophoblast
(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Gauster, Martin; Blaschitz, Astrid; Siwetz, Monika; Huppertz, Berthold
Besides microfilaments and microtubules, intermediate filaments are major components of the cytoskeleton. In epithelial cells intermediate filaments are formed by heterodimers of specific keratins, whose expression pattern highly depends on the type of epithelium and differentiation degree of the cell. During the process of blastocyst implantation and subsequent development of the human placenta a very specialized epithelium appears at the feto-maternal interface. Arising from the trophectoderm of the blastocyst, the epithelium-like layer surrounding the early embryoblast, different trophoblast subtypes differentiate. They either develop into polar cells fulfilling real epithelial functions, or apolar tumor-like cells invading the maternal uterine wall to adapt the maternal tissue to progressing pregnancy. Thus, the whole trophoblast population, with all its subtypes, can be considered as an epithelial compartment and hence expresses keratin filaments. However, differentiation of trophoblast into different phenotypes may be linked to remodeling of the cytoskeletal composition, depending on spatiotemporal requirements of the respective cells. Here, we focus on the keratin composition of different trophoblast subtypes, how these keratins are used in trophoblast research and what is kn
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Kinetics of infection and effects on the placenta of Chlamydophila abortus in experimentally infected pregnant ewes
(SAGE Publications, 2004-09-01) García de la Fuente, J. N.; Sánchez, J.; Gutiérrez Martín, C. B.; Rodríguez Ferri, E. F.; Salinas, J.; Martínez Cáceres, Carlos Manuel; Navarro Cámara, José Antonio; Ortega Hernández, Nieves; Buendía Marín, Antonio Julián; Anatomía y Anatomía Patológica Comparadas
A Chlamydophila abortus-induced abortion model was carried out on the basis of the experimental infection of ewes at day 75 of gestation. The infection induced abortions and the birth of weak lambs during the last 3 weeks of pregnancy. To study the kinetics of the infection in the placenta and in other organs, infected ewes were killed at 105, 120, and 130 days of gestation and also several days after abortion or parturition. Infected ewes developed a systemic infection that caused a mild and transient pneumonia and focal hepatitis. Pathologic changes were observed in placentas at 120 day of gestation, although the lesions varied between animals and even between placentomes of the same placenta. The first placental area infected was the maternal stroma and epithelium next to the intercaruncular areas, where neutrophilic response seemed to control the infection. A substantial degree of multiplication of C. abortus was then observed in the trophoblast cells of the placentome, periplacentomal choriallantoic membranes, and hilius, with an inflammatory exudate composed mainly of neutrophils, some macrophages, and very scarce lymphocytes. After abortion, the lesions affected the intercotyledonary areas of the aborted placentas, whereas in the uterus significant lymphocyte infiltration was observed, together with a rapid decrease of the C. abortus antigen in the degenerated caruncular tissues.
Open Access
lntermediate filament protein expression and sugar moieties in normal canine placenta
(Murcia : F. Hernández, 2000) Fernandez, P.E.; Barbeito, C.G.; Portiansky, E.L.; Gimeno, E.J.
In the female dog, the placenta is considered zonal, endotheliochorial and labyrinthic. The distribution of the intermediate filaments as well as the surface glycoproteins in the canine placenta are still unknown. The aim of the present study was to provide this information for further understanding of pathological conditions in the bitch. Samples were obtained from normal uterine horns at the end of gestation. Tissues were routinely fixed and stained. Monoclonal antibodies against cytokeratins, vimentin and desmin were used for immunohistochemical staining. UEA-1, PNA, RCA-1, SBA, DBA, WGA and ConA were used for the lectin histochemical staining. A computer morphometrical analysis was made. Statistical analysis was then accomplished. The results showed the maximun immunohistochemical percentage for vimentin in the supraglandular connective tissue, for pancytokeratin in the spongy zone and for desmin in miometrium. SBA showed the highest staining percentage in the gland cells of the spongy zone, while ConA was the highest in the syncytiotrophoblastic cells and gland cells of the deep glandular zone. The results obtained indicate that the lectin binding pattern is partially different from other animal species. On the contrary, the intermediate filament data coincide with similar observations from other mammals.
Open Access
Localization of human placental glucose transporter 1 during pregnancy. An immunohistochemical study
(Murcia : F. Hernández, 1996) Tadokoro, C.; Yoshimoto, Y.; Sakata, M.; Fujimiya, M.; Kurachi, H.; Adachi, E.; Maeda, T.; Miyake, A.
To elucidate the potential roles of glucose transporter 1 (GLUTl) in human placenta during pregnancy, we examined the localization of GLUTl in human placenta at various stages by immunohistochemistry with an anti-GLUT1 antibody by use of both light and electron microscopy. Specific staining for GLUTl was localized on the apical brush border and along the basal plasma membrane of the syncytiotrophoblasts. The staining at the apical side was more intense than that at the basal side during the early stages of gestation. In later gestational stages, however, the staining pattern at the apical side became blurred and the staining intensity at the basal side increased. The cytotrophoblasts, seen embedded in the basal part of the syncytiotrophoblasts, seemed to show immunoreactivity for GLUTl along the plasma membranes at the lightmicroscopic level. However, immuno-electron microscopic analysis with either pre- or post-embedding methods revealed that specific staining for GLUTl was hardly observed on the cytotrophoblasts, but the cytotrophoblasts were often surrounded by immunoreactive processes of syncytiotrophoblasts. The blood capillaries and erythrocytes in the stroma of placental villi were always immunoreactive for GLUTl throughout pregnancy. These findings suggest that GLUTl may play a vital role in human pregnancy.
Open Access
Maternal diabetes affects cell proliferation in developing rat placenta
(Editores F. Hernandez y Juan F. Madrid. Murcia, Universidad de Murcia, Departamento de Biologia Celular e Histologia, 2011) Zorn, T.M.T.; Zúñiga, M.; Madrid, E.; Tostes, R.; Fortes, Z.; Giachini, F.; San Martín, S.
Placentation starts with the formation of a spheroidal trophoblastic shell surrounding the embryo, thus facilitating both implantation into the uterine stroma and contact with maternal blood. Although it is known that diabetes increases the placental size and weight, the mechanisms responsible for this alteration are still poorly understood. In mammals, cellular proliferation occurs in parallel to placental development and it is possible that diabetes induces abnormal uncontrolled cell proliferation in the placenta similar to that seen in other organs (e.g. retina). To test this hypothesis, the objective of this work was to determine cell proliferation in different regions of the placenta during its development in a diabetic rat model. Accordingly, diabetes was induced on day 2 of pregnancy in Wistar rats by a single injection of alloxan (40 mg/kg i.v.). Placentas were collected on days 14, 17, and 20 postcoitum. Immunoperoxidase was used to identify Ki67 nuclear antigen in placental sections. The number of proliferating cells was determined in the total placental area as well as in the labyrinth, spongiotrophoblast and giant trophoblast cell regions. During the course of pregnancy, the number of Ki67 positive cells decreased in both control and diabetic rat placentas. However, starting from day 17 of pregnancy, the number of Ki67 positive cells in the labyrinth and spongiotrophoblast regions was higher in diabetic rat placentas as compared to control. The present results demonstrate that placentas from the diabetic rat model have a significantly higher number of proliferating cells in specific regions of the placenta and at defined developmental stages. It is possible that this increased cell proliferation promotes thickness of the placental barrier consequently affecting the normal maternal-fetal exchanges.