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1.
Bull Exp Biol Med ; 171(2): 254-257, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34173099

RESUMO

Atomic force microscopy is not very popular in practical health care, therefore, its potential is not studied enough, for example, in obstetrics when studying the "mother-placenta-fetus" system. Our study summarizes the possibilities of using atomic force microscopy for detection of various circulatory disorders and vascular changes at the microscopic level in the uterus (endometrium and myometrium), placenta, and umbilical cord in the main variants of obstetric and endocrine pathology. For instance, in the case of endocrine pathologies, changes in the form of stasis, sludge, diapedesis, ischemia, destruction and separation of endotheliocytes in villous blood vessels were found in the mother. The oxygen content in erythrocytes also naturally decreased in pathologies; poikilo- and anisocytosis were observed.


Assuntos
Microscopia de Força Atômica , Complicações na Gravidez/diagnóstico , Diagnóstico Pré-Natal/métodos , Adulto , Estudos de Casos e Controles , Vilosidades Coriônicas/irrigação sanguínea , Vilosidades Coriônicas/diagnóstico por imagem , Vilosidades Coriônicas/patologia , Vilosidades Coriônicas/ultraestrutura , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 1/diagnóstico por imagem , Diabetes Mellitus Tipo 1/patologia , Diabetes Gestacional/sangue , Diabetes Gestacional/diagnóstico , Diabetes Gestacional/diagnóstico por imagem , Diabetes Gestacional/patologia , Feminino , Feto/irrigação sanguínea , Feto/diagnóstico por imagem , Testes Hematológicos/métodos , Humanos , Relações Materno-Fetais , Microscopia Eletrônica de Varredura , Miométrio/diagnóstico por imagem , Miométrio/patologia , Miométrio/ultraestrutura , Placenta/irrigação sanguínea , Placenta/diagnóstico por imagem , Placenta/patologia , Placenta/ultraestrutura , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/diagnóstico por imagem , Pré-Eclâmpsia/patologia , Gravidez , Complicações na Gravidez/sangue , Complicações na Gravidez/diagnóstico por imagem , Complicações na Gravidez/patologia , Gravidez em Diabéticas/sangue , Gravidez em Diabéticas/diagnóstico , Gravidez em Diabéticas/diagnóstico por imagem , Gravidez em Diabéticas/patologia , Doenças da Glândula Tireoide/sangue , Doenças da Glândula Tireoide/diagnóstico , Doenças da Glândula Tireoide/diagnóstico por imagem , Doenças da Glândula Tireoide/patologia , Cordão Umbilical/irrigação sanguínea , Cordão Umbilical/diagnóstico por imagem , Cordão Umbilical/ultraestrutura , Útero/irrigação sanguínea , Útero/diagnóstico por imagem , Útero/ultraestrutura
2.
Placenta ; 103: 86-93, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33120050

RESUMO

Hypertension is one of the major clinical manifestations of preeclampsia. Vascular dysfunction is crucial for the occurrence and progression of hypertension. Exosomes are emerging as mediators of intercellular communication and can participate in angiogenesis. In this study, we hypothesize that umbilical cord plasma-derived exosomes from preeclamptic women (PE-uexo) impair vascular development by regulating endothelial cells. Here, umbilical cord plasma samples from women with normal pregnancies and matched preeclamptic patients were used to isolate circulating exosomes. Proliferation, Transwell and tube formation assays indicated that PE-uexo impaired the angiogenesis of human umbilical vein endothelial cells (HUVECs). On the basis of microarray analysis of HUVECs treated with PE-uexo or exosomes from women with normal pregnancies, we showed that the expression of 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1) was decreased in the PE-uexo-treated HUVECs. Furthermore, downregulation of HMGCS1 in HUVECs attenuated the proliferation and migration of these cells. Interestingly, HMGCS1 was decreased in P0 HUVECs from preeclamptic pregnancies compared with normotensive pregnancies. Together, these observations suggest that PE-uexo disrupts normal function in vascular endothelial cells by targeting HMGCS1, which may result in vascular disorders in the offspring.


Assuntos
Exossomos/fisiologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Hidroximetilglutaril-CoA Sintase/genética , Pré-Eclâmpsia , Cordão Umbilical/citologia , Adulto , Estudos de Casos e Controles , Células Cultivadas , Feminino , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Hidroximetilglutaril-CoA Sintase/metabolismo , Neovascularização Patológica/sangue , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/patologia , Pré-Eclâmpsia/fisiopatologia , Gravidez , Cordão Umbilical/ultraestrutura
3.
Biomaterials ; 185: 39-50, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30218835

RESUMO

Clinical application of a large variety of biomaterials is limited by the imperfections in storage technology. Perspective approaches utilizing low-temperature storage are especially challenging for multicellular structures, such as tissues, organs, and bioengineered constructs. Placenta, as a temporary organ, is a widely available unique biological material, being among the most promising sources of various cells and tissues for clinical and experimental use in regenerative medicine and tissue engineering. The aim of this study was to analyse the mechanisms of cryoinjuries in different placental tissues and bioengineered constructs as well as to support the viability after low temperature storage, which would contribute to development of efficient biobanking technologies. This study shows that specificity of cryodamage depends on the structure of the studied object, intercellular bonds, as well as interaction of its components with cryoprotective agents. Remarkably, it was possible to efficiently isolate cells after thawing from all of the studied tissues. While the outcome was lower in comparison to the native non-frozen samples, the phenotype and expression levels of pluripotency genes remained unaffected. Further progress in eliminating of recrystallization processes during thawing would significantly improve biobanking technologies for multicellular constructs and tissues.


Assuntos
Criopreservação/métodos , Placenta/citologia , Adolescente , Adulto , Alginatos/química , Âmnio/citologia , Âmnio/ultraestrutura , Materiais Biocompatíveis/química , Bancos de Espécimes Biológicos , Engenharia Biomédica , Células Cultivadas , Células Imobilizadas/citologia , Células Imobilizadas/ultraestrutura , Crioprotetores/química , Feminino , Humanos , Placenta/ultraestrutura , Gravidez , Engenharia Tecidual , Cordão Umbilical/citologia , Cordão Umbilical/ultraestrutura , Adulto Jovem
4.
Medisan ; 22(2)feb. 2018. tab, fig
Artigo em Espanhol | LILACS | ID: biblio-894676

RESUMO

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


Assuntos
Humanos , Cordão Umbilical/ultraestrutura , Neovascularização Fisiológica , Células Endoteliais/ultraestrutura , Achados Morfológicos e Microscópicos , Neovascularização Patológica , Epidemiologia Descritiva , Estudos Transversais , Biologia Celular
5.
Stem Cell Res ; 25: 166-178, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29154076

RESUMO

Stem cell therapy is considered a promising approach in the treatment of amyotrophic lateral sclerosis (ALS) and mesenchymal stem cells (MSCs) seem to be the most effective in ALS animal models. The umbilical cord (UC) is a source of highly proliferating fetal MSCs, more easily collectable than other MSCs. Recently we demonstrated that human (h) UC-MSCs, double labeled with fluorescent nanoparticles and Hoechst-33258 and transplanted intracerebroventricularly (ICV) into SOD1G93A transgenic mice, partially migrated into the spinal cord after a single injection. This prompted us to assess the effect of repeated ICV injections of hUC-MSCs on disease progression in SOD1G93A mice. Although no transplanted cells migrated to the spinal cord, a partial but significant protection of motor neurons (MNs) was found in the lumbar spinal cord of hUC-MSCs-treated SOD1G93A mice, accompanied by a shift from a pro-inflammatory (IL-6, IL-1ß) to anti-inflammatory (IL-4, IL-10) and neuroprotective (IGF-1) environment in the lumbar spinal cord, probably linked to the activation of p-Akt survival pathway in both motor neurons and reactive astrocytes. However, this treatment neither prevented the muscle denervation nor delayed the disease progression of mice, emphasizing the growing evidence that protecting the motor neuron perikarya is not sufficient to delay the ALS progression.


Assuntos
Esclerose Lateral Amiotrófica/terapia , Transplante de Células-Tronco Mesenquimais , Neurônios Motores/citologia , Superóxido Dismutase-1/genética , Cordão Umbilical/transplante , Esclerose Lateral Amiotrófica/enzimologia , Esclerose Lateral Amiotrófica/genética , Animais , Feminino , Humanos , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios Motores/metabolismo , Mutação Puntual , Superóxido Dismutase-1/metabolismo , Cordão Umbilical/citologia , Cordão Umbilical/metabolismo , Cordão Umbilical/ultraestrutura
6.
Med Sci Monit ; 22: 4542-4554, 2016 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-27885249

RESUMO

BACKGROUND The complex process by which lactation is initiated upon neonate delivery remains incompletely understood. Microvesicles (MVs) can transmit microRNAs (miRNAs) into recipient cells to influence cell function, and recent studies have identified miRNAs essential for mammary gland development and lactation. This study aimed to investigate the expression of lactation-related miRNAs in MVs isolated from human umbilical cord blood immediately after delivery. MATERIAL AND METHODS Umbilical cord blood samples were collected from 70 healthy pregnant women, and MVs were isolated through differential centrifugation and characterized by transmission electron microscopy, Western blotting, and nanoparticle tracking analysis. Lactation-related miRNAs were screened using bioinformatics tools for miRNA target prediction, gene ontology, and signaling pathway analyses. miRNA PCR arrays were used for miRNA expression analysis, and the results were validated by real-time PCR. Upon exposure of HBL-100 human mammary epithelial cells to MVs, MV uptake was examined by fluorescence confocal microscopy and b-casein secretion was detected by ELISA. RESULTS Spherical MVs extracted from umbilical cord blood expressed CD63 and had an average diameter of 167.0±77.1 nm. We profiled 337 miRNAs in human umbilical cord blood MVs and found that 85 were related to lactation by bioinformatics analysis. The 25 most differentially expressed lactation-related miRNAs were validated by real-time PCR. MV uptake by HBL-100 cells was after 4 h in culture, and significantly increased secretion of ß-casein was observed after 96 h from cells exposed to MVs (P<0.05). CONCLUSIONS Umbilical cord blood MVs contain many lactation-related miRNAs and can induce ß-casein production by HBL-100 cells in vitro. Thus, umbilical cord blood MVs may mediate secretion of ß-casein through miRNAs, thereby playing an important role in fetal-maternal crosstalk.


Assuntos
Micropartículas Derivadas de Células/metabolismo , Sangue Fetal/metabolismo , Lactação/fisiologia , MicroRNAs/biossíntese , Adulto , Micropartículas Derivadas de Células/genética , Células Cultivadas , Feminino , Humanos , MicroRNAs/genética , Gravidez , Cordão Umbilical/metabolismo , Cordão Umbilical/ultraestrutura
7.
Rom J Morphol Embryol ; 57(2): 383-90, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27516009

RESUMO

In recent years, identification of new sources of adult stem cells developed rapidly, pursuing to find easily available tissues, which will give rise to homogenous stem cells populations. Up to present, bone marrow-derived mesenchymal stem cells (BM-MSCs) are unanimously considered to fulfill the criteria for being used in clinical settings, but adipose stem cells, placental and umbilical cord stem cells, and other tissue-derived stem cells are making their way to being used at least in autologous transplantation. We isolated cellular populations from placental tissue and umbilical cord using the explants method. The placental (PL) and umbilical cord (UC)-derived cells were cultured and expanded in appropriate conditions for generation of stem cells. We assessed the stemness characteristics of the tissue-isolated cells and compared them to an established MSCs line. For this purpose, we determined the immunophenotype, morphological and ultrastructural characteristics, as well as functional abilities of PL- and UC-derived cells. Flow cytometric evaluation of cells revealed presence of CD90, CD73, and CD105 stem cells markers, while the cells were negative for CD34, CD45 and HLA-DR. Immunocytochemical staining showed that 100% of PL- and UC-derived cells are positive for vimentin and CD105 expression, while cytokeratin was revealed in less than 10% in both tissue-isolated cells. Morphological and ultrastructural characteristics of cells exposed analogous cellular size and intracellular organization, similar to MSCs, but detailed view of UC-derived cells by transmission electron microscopy (TEM) demonstrated presence of intercellular junctions-desmosomes, similar to epithelial cells. Both PL- and UC-derived cells confirmed their trilineage potential, being able to differentiate into adipocytes, osteoblasts, and chondrocytes in different proportions. Flow chamber in vitro assay was used to determine to what extent PL- and UC-derived cells are able to adhere to substrates (VCAM and ICAM) and we showed progressively decreased adhesion of both cellular types, inversely proportional to the generated shear stress. We may conclude that explants-isolated placental and umbilical cord cells are endowed with characteristics of both epithelial and mesenchymal stem cells, and purification procedures are additionally required for safe use of these cells in diverse clinical applications.


Assuntos
Separação Celular/métodos , Células Epiteliais/citologia , Células-Tronco Mesenquimais/citologia , Placenta/citologia , Cordão Umbilical/citologia , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Biomarcadores/metabolismo , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem da Célula/efeitos dos fármacos , Membrana Celular/metabolismo , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Imunofenotipagem , Molécula 1 de Adesão Intercelular/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Gravidez , Cordão Umbilical/ultraestrutura , Molécula 1 de Adesão de Célula Vascular/farmacologia , Vimentina/metabolismo
8.
Int. j. morphol ; 33(4): 1277-1281, Dec. 2015. ilus
Artigo em Inglês | LILACS | ID: lil-772308

RESUMO

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.


Assuntos
Humanos , Feminino , Gravidez , Caderinas/metabolismo , Diabetes Gestacional , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Cordão Umbilical/metabolismo , Cordão Umbilical/ultraestrutura , Imuno-Histoquímica , Microscopia
9.
Int J Low Extrem Wounds ; 14(2): 136-45, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25700709

RESUMO

Human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are multipotent, primitive, and have been widely used for skin tissue engineering. Their transdifferentiation is determined by the local microenvironment. In this study, we investigated the potential epidermal differentiation of UC-MSCs and the formation of epidermis substitutes in a 3-dimensional (3D) microenvironment, which was fabricated by UC-MSCs embedded into collagen-chitosan scaffolds (CCSs) combined with an air-liquid interface (ALI) culture system. Using fluorescence microscope, we observed that UC-MSCs were spindle-shaped and evenly distributed in the scaffold. Methyl thiazolyl blue tetrazolium bromide assay and Live/Dead assay indicated that the CCSs have good biocompatibility with UC-MSCs. Immunohistochemistry and western blotting assay showed that UC-MSCs on the surface of the CCSs were positive for the epidermal markers cytokeratin 19 and involucrin at 14 days. In addition, hematoxylin-eosin staining indicated that multilayered epidermis substitutes were established. The constructed epidermis substitutes were applied to treat full-thickness wounds in rats and proved to promote wound healing. In conclusion, manipulating the 3D microenvironment is a novel method for inducing the epidermal differentiation of MSCs to engineer epidermal substitutes, which provides an alternative strategy for skin tissue engineering.


Assuntos
Transdiferenciação Celular , Células-Tronco Mesenquimais/citologia , Pele/lesões , Engenharia Tecidual/métodos , Cordão Umbilical/ultraestrutura , Cicatrização , Ferimentos e Lesões/terapia , Animais , Western Blotting , Diferenciação Celular , Células Cultivadas , DNA/administração & dosagem , Modelos Animais de Doenças , Epiderme/ultraestrutura , Humanos , Imuno-Histoquímica , Microscopia Eletrônica de Varredura , Ratos , Ratos Sprague-Dawley , Pele/patologia , Ferimentos e Lesões/patologia
10.
Appl Immunohistochem Mol Morphol ; 23(6): 430-2, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25390355

RESUMO

The embryonic origin of umbilical cord vestiges is well documented; however, their immunophenotype is unknown. This study was conducted to determine whether vitelline and allantoic remnants can be differentiated using immunohistochemical markers. All allantoic remnants were stained with p63 and were negative for CDX2, whereas the vitelline remnants stained with CDX2 and were negative for p63. An unexpected finding was a small number of morphologically ambiguous cases that stained with both markers in a complimentary manner. The term "hybrid" remnant is proposed for these remnants.


Assuntos
Alantoide/ultraestrutura , Cordão Umbilical/ultraestrutura , Membrana Vitelina/ultraestrutura , Alantoide/metabolismo , Biomarcadores/metabolismo , Fator de Transcrição CDX2 , Amarelo de Eosina-(YS) , Feminino , Expressão Gênica , Hematoxilina , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Coloração e Rotulagem , Cordão Umbilical/metabolismo , Membrana Vitelina/metabolismo
11.
Ultrastruct Pathol ; 37(3): 196-203, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23650992

RESUMO

The umbilical cord contains mucinous connective tissue, called Wharton's jelly. It consists of stromal cells, collagen fibers, and amorphous ground substances composed of proteoglycan. Recently, these stromal cells have been redefined as a new cell therapy source, named human umbilical cord-derived mesenchymal stromal cells (hUCMSCs). However, there are few studies on the ultrastructural features and immune-phenotypic characteristics of isolated hUCMSCs and comparisons with the cells found in original cord tissues. In this study, the authors describe and compare the phenotypic characteristics of hUCMSCs with cells in the umbilical cord in order to know the kinds of cells and ultrastructural changes. Isolated hUCMSCs showed similar ultrastructure with few structural differences from in situ stromal cells, and they are relatively homogenous and well-developed mesenchymal cells that demonstrate a myofibroblastic phenotype.


Assuntos
Imunofenotipagem , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/ultraestrutura , Cordão Umbilical/imunologia , Cordão Umbilical/ultraestrutura , Geleia de Wharton/imunologia , Geleia de Wharton/ultraestrutura , Biomarcadores/análise , Separação Celular/métodos , Células Cultivadas , Feminino , Humanos , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Miofibroblastos/imunologia , Miofibroblastos/ultraestrutura , Fenótipo , Gravidez , Cordão Umbilical/citologia , Geleia de Wharton/citologia
12.
Med Mol Morphol ; 46(2): 77-85, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23446359

RESUMO

Chorangiosis is microscopically designated as more than ten terminal capillaries within the villous stroma of the placenta and is mostly related to chronic fetal hypoxia. However, the histogenetic relationship between increased number of terminal villous capillaries and chronic hypoxia has not yet been clarified. Of 665 placentas histologically examined at Saitama Medical University from 2003 to 2010, chorangiosis was found in 58 cases (8.7 %), which were mostly more than 35 gestational weeks. In addition, low birth weight (less than 2,500 g) infants (74.1 %) and those who suffered from cardiac anomalies, chromosome anomalies, and single umbilical artery comprised 32.7 % of cases. Placental lesions were associated with chorangiosis involved in infarct (46.6 %), intervillous thrombosis (20.7 %), and marginal hemorrhages (22.4 %). Scanning electron microscopic studies showed narrowing of vessel ostium and disorders of endothelium in the umbilical cord vessel complicated by chorangiosis. Furthermore, in transmission electron microscopic observation, not only the chorionic villi had multiple enlarged vessels within the villous stroma, but we also found that new capillaries were formed by angiogenesis with endothelial cells derived from fibroblasts under the chronic hypoxic state.


Assuntos
Vilosidades Coriônicas/ultraestrutura , Microscopia Eletrônica de Varredura , Neovascularização Patológica/fisiopatologia , Placenta/ultraestrutura , Capilares/fisiopatologia , Capilares/ultraestrutura , Vilosidades Coriônicas/fisiopatologia , Feminino , Hipóxia Fetal/fisiopatologia , Fibroblastos/patologia , Humanos , Hipóxia/complicações , Hipóxia/patologia , Lactente , Masculino , Neovascularização Patológica/complicações , Placenta/fisiopatologia , Gravidez , Cordão Umbilical/fisiopatologia , Cordão Umbilical/ultraestrutura
13.
Curr Stem Cell Res Ther ; 8(1): 60-72, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23270628

RESUMO

Human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs) are considered as a remarkable and promising stem cell source to be potentially used in cellular therapies. While no graft rejection has been reported in the recipient organism even in xeno-transplantation studies, attenuate tumor cell growth and gene transfers have been experimentally shown. In this study, we have demonstrated a reliable, reproducible and efficient cryopreservation method of hUCS-MSCs resulting in one of the highest cell survival rates reported so far. Conventional, computer-controlled multistep slow freezing (MSSF), and vitrification methods were comparatively tested using cell permeable [dimethylsulfoxide (DMSO), ethylene glycol] and impermeable [trehalose, sucrose, hydroxyethyl starch (HES), human serum albumin] cryoprotectant agents (CPAs). After determining the ice nucleation point for each solution, latent heat evolution was suppressed during freezing, followed by a cooling process to -40°C at 1°C/min or 0.3°C/min. The efficiency of the cryopreservation techniques used was determined by cell viability and proliferation assays, the expression of cell surface markers, cytoskeletal proteins and chromosome alignments. The cell survival rate was found to be highest (87 ± 5%) by MSSF with sucrose (0.1 M) +DMSO (10%) at 1°C/min freezing rate. In this group, no significant difference was noted before and after the cryopreservation in cell morphology, cytokeratin, vimentin, and α-smooth muscle actin profiles and the expressions of CD105, CD90, CD73, CD29 and HLA-DR. Second highest cell survival ratio (85 ± 6%) was obtained in DMSO (10%) alone at 1°C/min freezing rate. Interestingly, poor (18 ± 15%) cell survival rates were obtained after vitrification. Cumulatively, results indicated that MSSF favors the other freezing protocols with an addition of sucrose or DMSO alone depending on the freezing rate used.


Assuntos
Bancos de Espécimes Biológicos , Crioprotetores , Células-Tronco Mesenquimais/citologia , Células Estromais/citologia , Cordão Umbilical/citologia , Biomarcadores , Engenharia Celular , Criopreservação , Feminino , Citometria de Fluxo , Humanos , Técnicas Imunoenzimáticas , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/ultraestrutura , Gravidez , Células Estromais/metabolismo , Células Estromais/ultraestrutura , Cordão Umbilical/metabolismo , Cordão Umbilical/ultraestrutura
14.
Microsc Res Tech ; 75(10): 1445-51, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22718425

RESUMO

It is known that pre-eclampsia affects the structure of the umbilical cord including changes in diameter and wall thickness. In this work, the morphological changes of umbilical cords associated with pre-eclampsia were investigated using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The SEM images showed the overall structural changes in the umbilical cord, and the AFM imaged the surface of the cord in the nanometer range. The amount of Wharton's jelly was reduced in the cords of pre-eclampsia patients and it was holed along the boundary. Compared to a normal pregnancy, the surface of a pre-eclampsia cord was relatively smooth. In all components (Wharton's jelly, veins, and arteries), the values for surface roughness, Sa (average value of the roughness), Sq (root mean square), and Sz (peak to peak value), were smaller than those of the control (P < 0.05). Especially, the values for Sa of veins were ~fourfold less than those of the controls (P < 0.05). In pre-eclamptic cords, the amount of elastin in veins was increased while that of the artery was decreased.


Assuntos
Pré-Eclâmpsia/patologia , Cordão Umbilical/patologia , Cordão Umbilical/ultraestrutura , Feminino , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Gravidez
15.
Ginekol Pol ; 83(11): 858-61, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23379196

RESUMO

INTRODUCTION: The actual rate of multiple pregnancies is significantly larger than that observed during labor due to the fact that in the course of pregnancy intrauterine death of one or more fetuses may occur. CASE PRESENTATION: A twenty-six old woman (GII, PII) reported to hospital in 26 weeks of DC/DA twin gestation complicated by intrauterine death of one fetus in the second trimester of a spontaneous pregnancy. Pregnancy ended at term with vaginal birth of a single live fetus. After birth, entanglement of the fetal umbilical cord around the leg of the dead fetus was discovered. It was the most probable cause of death. CONCLUSIONS: Conservative management is preferred in case of intrauterine demise of one of the fetuses in DC/DA twin pregnancy. One of the reasons of fetal death may be entanglement of the umbilical cord around fetal small parts. The time of fetal death can be determined on the basis of the length of the thigh bone (Femur Length - FL).


Assuntos
Morte Fetal/diagnóstico por imagem , Doenças Fetais/diagnóstico por imagem , Gravidez de Gêmeos , Cordão Umbilical/ultraestrutura , Adulto , Evolução Fatal , Feminino , Doenças Fetais/patologia , Humanos , Recém-Nascido , Gravidez , Segundo Trimestre da Gravidez , Diagnóstico Pré-Natal/métodos , Ultrassonografia Pré-Natal , Cordão Umbilical/patologia
16.
Pesqui. vet. bras ; 30(10): 897-902, out. 2010. ilus
Artigo em Inglês | LILACS | ID: lil-567925

RESUMO

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.


Assuntos
Animais , Contagem de Células Sanguíneas/métodos , Contagem de Células Sanguíneas/veterinária , Cordão Umbilical/citologia , Cordão Umbilical/ultraestrutura , Bovinos
17.
Eur J Obstet Gynecol Reprod Biol ; 144 Suppl 1: S108-13, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19304367

RESUMO

The umbilical cord and its constituent tissues: an outer layer of amnion, porous Wharton's jelly, two umbilical arteries, and one umbilical vein, are designed to protect blood flow to the fetus during a term pregnancy. The outer amnion layer may regulate fluid pressure within the umbilical cord. The porous, fluid filled Wharton's jelly likely acts to prevent compression of the vessels. Blood flow is regulated by smooth muscle surrounding the arteries that is intermingled with a collagen based extracellular matrix (ECM). Doppler ultrasound measurements of blood flow within the umbilical cord, and at specific sites within the developing fetus, provide evidence of impaired blood flow in conditions such as preeclampsia. Mechanosensory communication between cells and the extracellular matrix (ECM) may likely result in cords possessing abnormal physical dimensions, impaired hemodynamics, and altered composition within the umbilical cord tissues. Few studies have explored the biomechanics of the intact umbilical cord, with its constituent tissues, from normal pregnancies or abnormal pregnancies, maternal or fetal complications. Here, alterations in the umbilical cord are reviewed concerning anatomical abnormalities, disease, or chromosomal alterations using sonography, Doppler ultrasound, histology, and biomolecular and biochemical analyses. This paper considers how current knowledge of the umbilical cord and its constituent tissues can be used to infer biomechanical function. In addition, the mechanical consequences of structural abnormalities and altered tissue structure or composition are discussed with a specific focus on preeclampsia.


Assuntos
Cordão Umbilical/fisiologia , Fenômenos Biomecânicos , Feminino , Humanos , Gravidez , Complicações na Gravidez/fisiopatologia , Fluxo Sanguíneo Regional , Cordão Umbilical/anatomia & histologia , Cordão Umbilical/ultraestrutura
18.
Int. j. morphol ; 26(3): 615-621, Sept. 2008. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-556721

RESUMO

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.


Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Placenta/anatomia & histologia , Placenta/embriologia , Placenta/ultraestrutura , Recém-Nascido Pequeno para a Idade Gestacional/crescimento & desenvolvimento , Recém-Nascido Pequeno para a Idade Gestacional/fisiologia , Recém-Nascido Pequeno para a Idade Gestacional/sangue , Recém-Nascido/crescimento & desenvolvimento , Recém-Nascido/sangue , Cordão Umbilical/anatomia & histologia , Cordão Umbilical/irrigação sanguínea , Cordão Umbilical/ultraestrutura , Saúde das Minorias Étnicas , Técnicas Histológicas/métodos
19.
Radiol. bras ; 41(3): 159-162, maio-jun. 2008. ilus, tab
Artigo em Inglês, Português | LILACS | ID: lil-486628

RESUMO

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.


Assuntos
Humanos , Feminino , Gravidez , Cistos , Cordão Umbilical/anormalidades , Cordão Umbilical/ultraestrutura , Doenças Fetais/diagnóstico , Brasil , Estudos Retrospectivos
20.
Reprod Fertil Dev ; 20(4): 537-45, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18462617

RESUMO

Evidence from several sources supports a close phylogenetic relationship between elephants and sirenians. To explore whether this was reflected in similar placentation, we examined eight delivered placentae from the Amazonian manatee using light microscopy and immunohistochemistry. In addition, the fetal placental circulation was described by scanning electron microscopy of vessel casts. The manatee placenta was zonary and endotheliochorial, like that of the elephant. The interhaemal barrier comprised maternal endothelium, cytotrophoblasts and fetal endothelium. We found columnar trophoblast beneath the chorionic plate and lining lacunae in this region, but there was no trace in the term placenta of haemophagous activity. The gross anatomy of the cord and fetal membranes was consistent with previous descriptions and included a four-chambered allantoic sac, as also found in the elephant and other afrotherians. Connective tissue septae descended from the chorionic plate and carried blood vessels to the labyrinth, where they gave rise to a dense capillary network. This appeared to drain into shorter vessels near the chorionic plate. The maternal vasculature could not be examined in the same detail, but maternal capillaries ran rather straight and roughly parallel to the fetal ones. Overall, there is a close resemblance in placentation between the manatee and the elephant.


Assuntos
Placentação/fisiologia , Prenhez , Trichechus inunguis/fisiologia , Animais , Animais Recém-Nascidos , Membranas Extraembrionárias/anatomia & histologia , Membranas Extraembrionárias/irrigação sanguínea , Feminino , Masculino , Placenta/anatomia & histologia , Placenta/fisiologia , Placenta/ultraestrutura , Circulação Placentária/fisiologia , Gravidez , Cordão Umbilical/ultraestrutura
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