Clasificación morfológica de células endoteliales de venas del cordón umbilical humano en imágenes digitales de cultivos in vitro / Morphological classification of endothelial cells of veins from the human umbilical cord in digital images of in vitro cultures

Se realizó un estudio descriptivo y transversal desde julio hasta octubre de 2017, por especialistas de la Universidad de Oriente y de la Universidad de Sao Paulo, Brasil, para analizar desde el punto de vista morfológico células endoteliales de venas del cordón umbilical humano, presentes en imágenes digitales de cultivos in vitro 2D, tratadas con la ß2GPI. . Se propuso la clasificación supervisada celular considerando 3 clases: circulares, deformadas alargadas y deformadas poco alargadas, según los coeficientes de formas elíptico y circular, todo lo cual permitió identificar formas celulares relevantes. Para comparar los resultados de las muestras de control y las tratadas, se calcularon los intervalos de confianza para cada una de las clases, con un nivel de confianza de 95 por ciento. Se concluye que el análisis de las alteraciones morfológicas in vitro puede ser utilizada en cultivos 2D precoces (de 24 y 48 horas) para la cuantificación de la angiogénesis

A descriptive and cross-sectional study was carried out from July to October, 2017, by specialists of the Oriente University and the University of Sao Paulo, Brazil, to analyze from the morphological point of view endothelial cells of the human umbilical cord veins, which were present in digital images of 2D in vitro cultures, treated with the ß2GPI. The cellular supervised classification was proposed considering 3 classes: circular, distorted elongated and distorted not very elongated, according to the coefficients of elliptic and circular shapes, all that allowed to identify outstanding cellular forms. To compare the results of the control and treated samples, the intervals of confidence were calculated for each of the classes, with a 95 percent level of confidence. It was concluded that the analysis of the morphological disorders in vitro can be used in early 2D cultures (24 and 48 hours) for the quantification of the angiogenesis

Expression of PECAM-1 and E-Cadherin in the Umblical Cords of Gestational Diabetic Mothers / Expresión de PECAM-1 y E-cadherina en los Cordones Umbilicales de Madres con Diabetes Gestacional

The purpose of this study is to examine the changes in the umbilical cord in women diagnosed with gestational diabetes mellitus In this study, as a control group human placental tissues from normotensive pregnancies was collected from diabetic women at 28­35 weeks of gestation. Gestational diabetes (n= 20) and normal umbilical cord (n= 20) for a total of 40 units were received.GDM groups compared to the control group was significantly higher values was detected (p<0.01). In GDM group, light microscopy showed erosion of the endothelium and complete rupture of theumbilicalvessels resulting in extravasation of blood within Wharton's jelly. it was observed that the cytoplasmic fragments and cell infiltration of the spill to the subepithelial layer of apoptotic cell PECAM-1 positive reaction showed. E-Cadherin in endothelial side surface of diabetes group showed weak expression in the nucleus and showed positive reaction in smooth muscle.

El objetivo fue examinar los cambios que presenta el cordón umbilical de mujeres con diagnóstico de diabetes mellitus gestacional (DMG). Se incluyeron en el grupo control muestras de tejidos placentarios humanos de embarazos normotensos y de mujeres diabéticas de entre 28­35 semanas de gestación. Las muestras se divieron en cordones umbilicales con cambios de DMG (n= 20) y cordones umbilicales normales (n= 20), constituyendo un total de 40 muestras. El grupo de DMG, en comparación con el grupo control, presentó valores significativamente más elevados (p<0,01). En el grupo de DMG, la microscopía óptica demostró la erosión del endotelio y la ruptura completa de los vasos umbilicales, resultando en la extravasación de sangre dentro de la gelatina . Se observaron fragmentos citoplasmáticos e infiltración celular de la capa subepitelial de células apoptóticas mostró una reacción positiva a PECAM-1. En el grupo de DMG, la E-cadherina de la superficie lateral endotelial mostró una expresión débil en el núcleo y una reacción positiva en el músculo liso.

Ultrastructural characterization of bovine umbilical cord blood cells / Caracterização ultra-estrutural das células sanguíneas do cordão umbilical bovino

The umbilical cord blood (UCB) is an important source of pluripotent stem cells, which motivated researches on ontogeny and transplantation. The morphological characterization of umbilical cord cells is the first step to establish subsequent experiments on these areas. Although some information on humans can be found, no data on UCB is available for bovines. Therefore, this work is the first attempt to conduct an ultrastructural characterization of bovine umbilical cord blood. Blood was collected from the umbilical cord of twenty fetuses by punction of the umbilical vein. Samples were processed for whole leucocytes observation by centrifugation and the buffy coat was collected. Cells were washed and pelleted and prepared according to the standard protocol of the transmission electron microscopy. The presence of cells with morphologic characteristics compatible with the precursors from the erythrocytic, neutrophilic, eosinophilic, basophilic, and lymphocytic lineages was observed. Atypical cells with peculiar morphological features, strongly similar to apoptotic cells, were seen. Bovine neutrophils with three types of cytoplasmic granules were also found in the blood. The ultrastructural characteristics of observed bovine UCB cells where similar to those found in other species, suggesting that bovines could possibly constitute an experimental model for approaches on UCB cells research.

O sangue de cordão umbilical (SCU) é uma importante fonte de células progenitoras pluripotentes, que motiva pesquisas em ontogenia e transplantes. A caracterização morfológica das células de cordão umbilical é o primeiro passo para se estabelecer experimentos subsequentes nessas áreas. Embora algumas informações sobre SCU em humanos possam ser encontradas, não existe nenhuma informação disponível sobre elas em bovinos. Portanto, este trabalho é a primeira tentativa de se conduzir uma caracterização ultra-estrutural do sangue de cordão umbilical bovino. O sangue foi coletado do cordão umbilical de 20 fetos por punção da veia umbilical. As amostras foram processadas para observação dos leucócitos totais por centrifugação pela coleta do botão leucocitário. As células foram lavadas, peletizadas e preparadas de acordo com protocolo padrão para microscopia eletrônica de transmissão. A presença de células com características morfológicas compatíveis com precursores das linhagens eritrocítica, neutrofílica, eosinofílica, basofílica e linfocítica foram observadas. Células atípicas com características morfológicas peculiares muito semelhantes a células em apoptose foram observadas. No sangue do cordão umbilical também foi encontrado neutrófilos bovinos apresentando três tipos de grânulos citoplasmáticos. As características ultraestruturais do SCU bovino foram semelhantes às encontradas em outras espécies, sugerindo que esta espécie possa servir como modelo experimental para abordagens em pesquisa sobre sangue de cordão umbilical.

Ultrastructural study of human umbilical cord blood Buffy coat and Wharton's Jelly cells

Human umbilical cord [UC] has been a tissue of increasing interest in recent years. Umbilical cord blood [UCB] was used in the treatment of several hematological diseases. New potential uses of cord blood in nonhematopoietic applications are now proposed. Lately, the Wharton's jelly [WJ], the embryonic mucuos connective tissue of the UC, with its mesenchymal stromal cells represents a promising source of cells for several regenerative therapies. This study aims to highlight the morphological ultrastustural characteristics of the UCB buffy coat and the WJ stromal cells in situ. Tiny specimens from the UCB buffy coat and the WJ were double-fixed with glutaraldehyde and osmium tetroxide, dehydrated, and embedded in epoxy resin. Ultrathin sections were cut and stained with uranyl acetate and lead citrate. Transmission electron microscopic examination [TEM] of the UCB buffy coat cells showed immature cellular constituents and few changes in the ultrastructural morphology of cord blood platelets. These ultrastructural changes may clarify the specific cultural, immunological and regenerative properties attributed to cord blood in comparison to bone marrow or peripheral blood. Wharton's Jelly TEM showed stromal cells exhibiting morphological resemblance with both fibroblasts and smooth muscle cells. In addition, cells resembling stellate cells exhibiting intracytoplasmic lipid globules and ultrastructural properties related to protein synthesis were discerned. Also, oval cells with primitive nuclear and cytoplasmic features pointing to a subpopulation with stem cell potency were seen. We suggest that the WJ contains several distinct populations of stromal cells with different potentials including a primitive subpopulation. We propose that the nomenclature of these cells should not only be linked to their morphological appearance but also to their specific functional properties. Extensive in situ and in vitro molecular, immunohistochemical and immuno electron microscopic studies are necessary to clearly identify these cell populations with different potentials

Características morfológicas y morfométricas de la placenta de término, en recién nacidos pequeños para la edad gestacional (PEG) en la ciudad de Temuco-Chile / Morphometric and morphological characteristics of the placenta at term, in small for gestational age newborns (SGA) in the city of Temuco-Chile

El metabolismo placentario, el intercambio de sustancias y la producción de hormonas son funciones vitales de la placenta para mantener y promover el desarrollo normal del feto. Existen factores de riesgo que alteran este patrón en el caso del retardo del crecimiento intrauterino, cuyo resultado será un recién nacido (RN) pequeño para la edad gestacional (PEG) que presentará una mayor morbilidad, crecimiento físico e intelectual comprometido y una mayor probabilidad de desarrollar durante la vida adulta diferentes patologías. Los objetivos del presente trabajo son: 1. Reconocer las diferencias en los parámetros morfométricos, como el área de las vellosidades, el área de los vasos, el número de vasos y el área del sinciciotrofoblasto de las placentas de PEG en relación con placentas de recién nacidos AEG y 2. Relacionar el diagnóstico neonatal de PEG con las características morfométricas. Se utilizaron 25 placentas de término (37-42 semanas), 12 de recién nacidos adecuados a la edad gestacional (AEG), y 13 de recién nacidos pequeños para la edad gestacional (PEG). Las muestras fueron obtenidas de la maternidad del Hospital Hernán Henríquez Aravena de Temuco, IX Región Chile. De cada placenta se tomaron dos segmentos pericordonales, desde la placa subcorial hasta la placa basal y luego fueron fijadas en formalina tamponada al 10 por ciento. Las técnicas histológicas utilizadas fueron H-E azul de Alcián, Tricrómico de Masson, PAS-Hematoxilina y PAS-Diastasa. El área de las vellosidades mostraron diferencias significativas entre el grupo control (AEG) y el grupo PEG con p = 0,0194. En el grupo de PEG el área de los vasos fue significativamente mayor, con un valor de 234,05 i,m² en comparación con el grupo control cuyo promedio fue de 150.99 lm² (p = 0,0001). El número de vasos sanguíneos por vellosidad libre no mostró diferencias significativas. En relación con el área del sinciciotrofoblasto la diferencia no resultó ser significativamente ...

The placental metabolism, the exchange of substances and the production of hormones are vital functions of the placenta to maintain and promote the normal development of the fetus. There are risk factors that disrupt this pattern in the case of intrauterine growth retardation, whose outcome will be a small for gestational age (SGA) newborn having a higher morbidity, physical and intellectual growth pledged and greater probability of develop different pathologies during adulthood. The aims of this study are: 1 .-recognize the morphometric parameters differences as the area of the villi, the area of the vessel, the number of vessels and the area of placental syncytiotrophoblast SGA in connection with placentas of newborns AGA and 2.- relate the diagnosis of neonatal SGA with morphometric characteristics. We used 25 placenta at term (37-42 weeks), 12 newborns appropriate to the gestational age (AEG), and 13 small for gestational age infants (SGA). The samples were obtained from the Maternity Hospital Hernán Henriquez Aravena of Temuco, Chile IX Región. In each placenta two segments were taken from the subchorionic plate to the basal plate and then were fixed in 10 percent formalin buffered. The histological techniques used were H- E Alcián blue, Masson's Trichromic, Pas-hematoxylin Pas-diastase. The area of the villi showed significant differences between the control group (AEG) and the PEG group with p = 0.0194. In the group of PEG the area of vessels was significantly higher, with a value of 234.05 mm² compared with the control group whose average was 150.99 mm² (p = 0.0001). The number of blood vessels for free villi sampling not significant differences. Regarding the area of syncytiotrophoblast the difference was not significantly (p = 0.1410). In conclusion it was determined that PEG newborns placenta showed significant differences at the blood vessel area and free chorial villi area in relation to the AEG placenta.

Correlação entre o achado ultra-sonográfico isolado de cisto de cordão umbilical e anomalias fetais / Correlation between isolated sonographic finding of umbilical cord cyst and fetal anomalies

OBJETIVO: Correlacionar o achado ultra-sonográfico isolado de cisto de cordão umbilical com anomalias fetais, como cromossomopatias e alterações estruturais. Segundo a literatura médica, as implicações clínicas do achado ultra-sonográfico de cisto de cordão nos segundo e terceiro trimestres de gestação estão bem estabelecidas, entretanto, quando no primeiro trimestre, o significado ainda permanece controverso. MATERIAIS E MÉTODOS: Foi realizado estudo retrospectivo de gestantes da população geral, consecutivas, com fetos únicos e vivos, que apresentavam apenas o achado de cisto de cordão umbilical, num período de dez anos (1996-2006). Em todos os casos foram realizados exames ultra-sonográficos para o rastreamento de anomalias fetais após o diagnóstico de cisto de cordão. Os recém-nascidos e o cordão umbilical foram examinados após o parto para se verificar a presença de anomalias. RESULTADOS: Foram estudados nove casos que apresentavam cisto de cordão umbilical como único achado, sem outros marcadores ultra-sonográficos de anomalias fetais. Detectaram-se dois casos no primeiro trimestre de gestação e sete nos segundo e terceiro trimestres. Dois casos foram submetidos a estudo citogenético fetal, por meio de amniocentese. Nenhum recém-nascido apresentou anomalias estruturais ou aneuploidia. CONCLUSÃO: O achado ultra-sonográfico isolado de cisto de cordão umbilical não significou aumento de risco para anomalias estruturais ou aneuploidias.

OBJECTIVE: To correlate the isolated sonographic finding of umbilical cord cyst with fetal anomalies such as chromosomopathies and structural changes. According to the medical literature, the clinical implications of the sonographic finding of umbilical cord cyst in the second and third trimesters of pregnancy are well established; however, the meaning of this finding in the first trimester still remains controversial. MATERIALS AND METHODS: A retrospective study was developed with consecutive, pregnant women with single living fetuses presenting with umbilical cord cyst as an isolated finding, over a 10-year period (1996-2006). Ultrasound studies were performed in all cases for screening of fetal anomalies after the diagnosis of umbilical cord cyst. Neonates and umbilical cords were evaluated after delivery for the presence of abnormalities. RESULTS: Nine cases presenting umbilical cord cyst as a sole finding with no other sonographic marker for fetal abnormality were evaluated. Two cases were detected in the first pregnancy trimester and seven cases in the second and third trimesters. Fetal cytogenetic study was done by means of amniocentesis in two cases. No newborn presented with structural anomalies or aneuploidy. CONCLUSION: Isolated sonographic finding of umbilical cord cyst did not imply increased risk for fetal structural anomalies or aneuploidies.

Estudio ultraestructual de placenta y cordón umbilical en embarazos complicados con oligohidramnios severo idiopático / Ultraestructural study of placenta and umbilical chord in pregnancy complicated by idiopathic oligohydramnios

En la ultraestructura de los tejidos estudiados se aprecian alteraciones que son comunes a otras enfermedades, como compromiso en el riego sanguíneo uteroplacentario. La asociación entre retardo en el crecimiento intrauterino y oligohidramnios idiopático, parece estar más en relación con hipoxia favorecida por alteraciones en la arquitectura de los vasos sanguíeos, principalmente la íntima, y además por el escaso número y tipo de organelos citoplasmáticos, como mitocondrias, cuyo papel es la producción de ATP y fosforilación oxidativa. Las alteraciones de endotelio y sinciciotrofoblasto pueden afectar la pruducción y depuración del líguiqdo amniótico y favorecer isquemia, con mayor disminución del volumen amniótico. Esta entidad favorece las muertes perinatales, principalmente por compromiso del cordón umbilical, e incrementa la morbilidad feral.