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PubMed2003

Peroxisome proliferator-activated receptor isoform expression changes in human gestational tissues with labor at term.

Peroxisome proliferator-activated receptors (PPARs) are a family of nuclear receptors that are involved in lipid metabolism, differentiation, proliferation, cell death, and inflammation. Three subtypes have been identified: PPAR-alpha, -delta, and -gamma. We have previously shown presence of PPAR-gamma mRNA in the amnion, choriodecidua, and placenta, and its level of expression was unchanged with labor. To evaluate whether PPAR-alpha and -delta subtypes are present in intrauterine tissues, placentae were obtained from women at term after spontaneous vaginal delivery (TSL; n = 15) and elective caesarean section before labor (TNL; n = 15). Northern blot analyses were used to evaluate the mRNA for PPARs. Activities of PPARs were assessed using JEG3 choriocarcinoma cells transfected with a PPAR-response element reporter construct (pTK-PPREx3-luc) and treated with PPAR ligands. The PPAR-gamma-specific ligand rosiglitazone induced PPAR response element (PPRE)-mediated activity in a concentration-dependent manner, whereas the PPAR-gamma-specific irreversible inhibitor GW9662 fully inhibited this induction. However, GW9662 only partially inhibited 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2)-induced luciferase activity, suggesting that 15d-PGJ2 may also activate either of the other isoforms. PPAR-alpha and -delta are expressed in the amnion, choriodecidua, and placental villous tissues. In the amnion, although for PPAR-alpha no significant difference in expression was observed with labor, PPAR-delta expression increased significantly (p < 0.001). In the choriodecidua, expression of PPAR-alpha declined with labor (p < 0.01), whereas, as in the amnion, PPAR-delta expression increased (p < 0.05). In the placenta, both PPAR-alpha and -delta expression increased with labor (p < 0.005). The changes observed with labor suggest that regulation of PPAR expression and function may have roles to the mechanisms that maintain pregnancy or initiate labor.

Amnion↗
PubMed1994

Studies on the action of interleukin-1 on term human fetal membranes and decidua.

Recent studies have indicated the possible importance of cytokines in the onset of term and preterm labor. To examine this further, the effect of interleukin-1 beta (IL-1 beta), interleukin-1 alpha (IL-1 alpha), and interleukin-6 (IL-6) on prostaglandin output by dispersed cells from human amnion, chorion laeve, and decidua obtained at term (38-40 weeks gestation) was examined. During the first or second 24 h of culture no significant effect of these interleukins on prostaglandin output was observed. The reason for this apparent refractoriness was further investigated by studying the distribution of IL-1 receptors in frozen sections of undisrupted fetal membranes and decidua at term. Whole-tissue autoradiography indicated that receptors were present in chorion-decidua but not in amnion. By using emulsion autoradiography, IL-1 receptors were found in high concentrations in chorion laeve and were absent in amnion and at low levels in the decidua. These studies indicate that under normal circumstances in human pregnancy at term IL-1 did not stimulate prostaglandin production by dispersed cells. In the case of amnion, this may be due to the absence of receptors, and therefore it would appear that the IL-1 receptor must first be induced in this tissue before it can respond to this cytokine. Furthermore, although chorion laeve expresses the IL-1 receptor, dispersed cells from this tissue did not respond to the cytokine by increasing prostaglandin output.

Amnion↗
PubMed2000

Differential distribution of ERalpha and ERbeta mRNA in intrauterine tissues of the pregnant rhesus monkey.

Two estrogen receptor (ER) isoforms, ERalpha and ERbeta, have been described. However, no information is available in any species regarding the comparison of ERalpha and ERbeta levels in pregnant intrauterine tissues. We investigated 1) distribution of ERalpha and ERbeta mRNA in myometrium, amnion, choriodecidua, and placenta; 2) their abundance in intrauterine tissues at term not in labor (NIL) and in spontaneous term labor (STL); and 3) immunolocalization of ERalpha and ERbeta in pregnant rhesus monkey myometrium. Myometrium, amnion, choriodecidua, and placenta were obtained at cesarean section from monkeys in STL at 156-166 days gestational age (GA) (n = 4) and from control monkeys NIL at 140-152 days GA (n = 4). RT-PCR was conducted to determine ERalpha and ERbeta and glyceraldehyde-3-phosphate dehydrogenase mRNA abundance in four intrauterine tissues of the pregnant rhesus monkey. The cloned ERbeta PCR fragment was subjected to sequence analysis. ERalpha and ERbeta were localized in the myometrium by immunohistochemistry. We demonstrated that 1) rhesus monkey ERbeta shares >97% identity with human ERbeta in the region sequenced; 2) both ERs were expressed in myometrium, amnion, and choriodecidua but not in placenta in the current study; 3) ERalpha and ERbeta were differentially distributed in myometrium and amnion; 4) ERalpha and ERbeta were immunolocalized in myometrial smooth cells and smooth muscle and endothelial cells of the myometrial blood vessels. The biological significance of these quantitative differences in ER subtypes merits further study.

Amnion↗
PubMed1997

Tissue-specific expression of the relaxed conformation of plasminogen activator inhibitor-2 and low-density lipoprotein receptor-related protein in human term gestational tissues.

The relaxed conformation of plasminogen activator inhibitor-2 (PAIr) is formed during inactivation of the matrix-degrading enzyme urokinase plasminogen activator (uPA). The presence of PAIr in tissues, therefore, indicates the in situ inhibition of uPA-mediated proteolysis. In addition, PAIr functions as a ligand for the clearance receptor low-density lipoprotein receptor-related protein (LRP), thereby promoting internalization of receptor-bound uPA-PAIr complexes from the cell surface. The rapid internalization of receptor-bound, inactivated uPA has been suggested to be characteristic of invasive cell phenotypes. The aims of this study were to characterize the immunohistochemical localization of PAIr in human term gestational tissues (amnion, choriodecidua, and placenta) and to establish its co-expression with other components of the uPA cascade. The results obtained indicate that PAIr immunoreactivity was exclusively localized to amnion epithelial cells, with only minimal staining in the underlying chorion. PAIr immunoreactivity was not detectable in any of the trophoblastic tissues examined (villous and extravillous). The tissue-specific expression of PAIr immunoreactivity was not significantly altered in association with labor onset. uPA and PAI-2 staining was localized predominantly to amnion epithelial cells, underlying chorion, and trophoblast cells of villous and extravillous tissue. Amnion and trophoblasts of extravillous and chorionic tissue showed uPAR immunoreactivity, whereas staining in placenta was absent. Immunoreactive LRP was confined to trophoblasts of the chorion, and the villous and extravillous tissue. For the first time, localization of PAIr at the tissue level has been identified. The data obtained are consistent with the hypothesis that cells of invasive phenotype, although expressing all components of the uPA cascade, do not accumulate immunoreactive PAIr, because it is rapidly internalized from the cell surface. Conversely, cells of noninvasive phenotype will accumulate PAIr immunoreactivity only in the absence of LRP expression. We propose that the presence of PAIr and the absence of LRP at the cell surface are putative markers of noninvasive phenotypes.

Amnion↗
PubMedDate not supplied

Inducing proliferation of human amniotic epithelial (HAE) cells for cell therapy.

Probably because amnion is derived from the fetus and is exposed to the maternal immune system, human amniotic epithelial (HAE) cells do not express the HLA-A, -B, -C, or -DR antigens on their surfaces, suggesting that HAE cells do not induce rejection (immune reaction) after allotransplantation. And the amnion, like the placenta, is useless to the mother and child after birth. Therefore, HAE cells or tissues were expected to be suitable for allotransplantation. Because HAE cells produce large amounts of enzymes, amnion transplantation has been carried out in order to correct inborn errors of metabolism by supplementing lysosomal enzyme deficiencies. However, several problems remain before amnion allotransplantation can be accepted as effective. The HAE cell population is limited, because the maximum number of HAE cells obtainable from one donor is about 2 x 10(8) cells, and HAE cells proliferate poorly in in vitro culture. In this study, we aimed at increasing the HAE cell population in vitro. First, we investigated the effect of several cytokines on HAE cell proliferation and found that hepatocyte growth factor (HGF), epidermal growth factor (EGF), and transforming growth factor-beta stimulated it, whereas IL-6 and LIF inhibited it. Second, we investigated the effects of amniotic fluid on HAE cell proliferation and observed that IL-6 in amniotic fluid inhibits it. Then, to inhibit the dying of cells, we attempted to inhibit apoptosis (one mode of cell death). Treatment with caspase III inhibitor increased the cell viability of HAE cells by 20%.

Amnion↗
PubMed2003

Expression and localization of prostaglandin E synthase isoforms in human fetal membranes in term and preterm labor.

Increased prostaglandin (PG) synthesis by fetal membranes occurs at parturition. PGE(2) synthesis from arachidonic acid involves multiple enzymes and two isoforms of the terminal enzyme of this biosynthetic pathway, PGE synthase (PGES), were recently identified. Cytosolic PGES (cPGES) is identical to the heat shock protein 90 chaperone, p23, and is reportedly functionally coupled to constitutive PG endoperoxide H synthase-1. Microsomal PGES (mPGES) is inducible by proinflammatory cytokines such as IL-1 beta. We have studied expression and localization of both enzyme isoforms in human fetal membranes either at term or preterm, with or without labor. The cPGES was immunolocalized in the amnion epithelium, and associated with fibroblasts and macrophages in the choriodecidual layer, whereas mPGES was localized in the amnion epithelium as well as the chorion trophoblast. Both enzymes were found to be associated with lipid particles present in the amnion epithelium, which are more prevalent in term tissues. Western blot analysis of the amnion and choriodecidua showed no differences in amounts of either cPGES or mPGES at term or preterm, with or without labor, in either tissue with advancing gestation. It does not appear that expression of PGES is the rate-limiting step in PGE2 synthesis in fetal membranes at labor.

Amnion↗
PubMed1987

Amniotic membrane production of prostaglandin F2 alpha is reduced in dysfunctional human labor: results of in vivo and in vitro studies.

Mobilization of arachidonic acid from glycerophospholipids and prostaglandin (PG) release from fetal membranes were studied in women with dysfunctional labor in the absence of cephalopelvic disproportion or fetal malposition. Using superfusion of intact amnion and chorion, we found a slight decrease in PGE and a more significant decrease in PGF release by the amniotic side of the fetal membrane obtained from women with dysfunctional labor compared to that in women with normal labor (PGE: normal labor, 2992 pg/cm2.h; dysfunctional labor, 1846 pg/cm2.h; P less than 0.05; PGF: normal labor, 662 pg/cm2.h; dysfunctional labor, 204 pg/cm2.h; P less than 0.02). Release of both prostanoids was significantly greater from the amniotic side in tissues obtained after labor compared to that in prelabor tissue. Analysis of arachidonic acid (by gas liquid chromatography) and phospholipid content (by two-dimensional thin layer chromatography) confirmed metabolic disposal of arachidonic acid from the amnion after the onset of labor. However, no difference in either phospholipid or phospholipase A2-releasable arachidonic acid of individual phospholipid classes was found in amnion tissue from women with normal and dysfunctional labor, suggesting similar activities of phospholipase A2 in these two groups. The finding of decreased free and phospholipase A2-releasable arachidonic acid of the total lipid extract of the amnion of women with dysfunctional labor could suggest further metabolic exhaustion of the substrate or failure of liberation of this fatty acid from glycerophospholipids by enzymes other than phospholipase A2, such as phospholipase C or diacyl and monoacylglycerolipases.

Adult↗
PubMed1993

The dynamics of prostaglandin metabolism in human fetal membranes and decidua around the time of parturition.

To address whether prostaglandins (PGs) produced in the amnion can gain access to the myometrium, we used three in vitro systems to determine the characteristics of PG secretion, metabolism, and transfer in amnion, chorion, and decidua around the time of parturition. PG metabolism, measured in explant cultures or cytosol preparations, occurs predominantly by the enzyme PG dehydrogenase, which was highest in chorion and was 2- to 3-fold more active for PGE2 than PGF2 alpha. The activity increased significantly around the time of labor onset. The activity of PG-9-ketoreductase, which interconverts the E and F series of PGs, was 2-3 orders of magnitude less than that of PG dehydrogenase. Using a dual chamber perfusion apparatus, we demonstrated that similar amounts of PGE2 were secreted from the fetal (amnion) and maternal (chorio-decidua) surfaces, and this ratio did not change with labor. Using radiolabeled PGE2, radioactivity traversed full thickness membranes at the rate of 4%/h. Only approximately 12% of the transferred radioactivity remained as intact PGE2, and very little conversion to PGF2 alpha was detected. No changes in transfer were detected around the onset of labor. We conclude that it is unlikely that PGE2 produced in the amnion acts directly on the myometrium.

Amnion↗
PubMed2004

Human amniotic epithelial cells possess hepatocyte-like characteristics and functions.

Hepatocyte transplantation is expected to become a novel method for treatment of liver disease. However, many questions remain regarding this approach, especially concerning donor cells. To evaluate whether human amniotic epithelial cells can be used as a cell source for hepatocyte transplantation, hepatic gene expression and functions of human amniotic epithelial cells were analyzed. Reverse transcription-polymerase chain reaction analysis demonstrated that human amniotic epithelial cells expressed albumin, alpha(1)-antitrypsin, and other hepatocyte-related genes. Cultivated human amniotic epithelial cells demonstrated albumin production, glycogen storage, and albumin secretion consistent with the hepatocyte gene expression profile. In organ culture, the amnion secreted 30-fold larger amounts of albumin than human amniotic epithelial cells in monolayer culture. Moreover, in organ culture the amnion also secreted alpha(1)-antitrypsin. Following transplantation into mice, the amnion survived and secreted albumin. These observations suggest that transplantation of human amniotic epithelial cells and/or amnion could be novel therapeutic strategy for treatment of hepatic diseases, including alpha(1)-antitrypsin deficiency.

Albumins↗
PubMed2001

Ultrasound determination of chorionicity in twin pregnancy: accuracy and operator experience.

To evaluate the accuracy of ultrasound in early detection of chorionicity and amnionicity in twin pregnancies and evaluate the role of operator-experience, a prospective follow up of 87 consecutive twin pregnancies was performed. Chorionicity and amnionicity were determined by 17 different observers, divided in 2 groups on the basis of their experience, using a composite of US markers at the first US examination confirming a viable intrauterine twin pregnancy. Accuracy of prediction was determined for both groups by comparison with postnatal pathology. From the 87 pregnancies examined at 10.1 (minimum 5.5 weeks, maximum 26.0) weeks of gestation, pathology was available in 82 cases and a correct prediction of chorionicity was made in all but two cases, both being diagnosed as monochorionic while dichorionic by observers belonging the more experienced group. No monochorionic pregnancy was missed and amnionicity was correctly diagnosed in all cases, including two monoamniotic twins. We conclude that use of a composite of ultrasound markers for early detection of chorionicity and amnionicity is reliable in a clinical setting where physicians with a variable degree of experience perform ultrasound examination.

Abortion, Spontaneous↗
PubMed1986

Site variability in the solubility of collagen of human fetal membranes.

The extractability of collagen was examined in different sites of amnion and chorion from term deliveries. Sequential extraction with NaCl, acetic acid and CaCl2 showed that soluble collagen accounted for 1.5% of the proteins extracted. Saline extracted more collagen from the amnion than from the chorion. Acetic acid and CaCl2 extracted decreasing and increasing amounts of collagen from the amnion and chorion respectively. The concentration of collagen decreased linearly in the chorion as the rupture site was approached. The results indicate differences in the nature of collagen between amnion and chorion as well as in various sites in the latter.

Acetates↗
PubMed1991

Localization and endogenous concentration of endothelin-like immunoreactivity in human placenta.

The aim of this study was to investigate the localization of endothelin-like immunoreactivity (ET-IR) in human placenta, chorion and amnion and to compare the endogenous concentration of immunoreactive endothelin (ET) in these tissues before and after the onset of labour. ET-IR was detected in the endothelium of stem vessels in placental villi, as well as in decidual stromal cells in the basal maternal plate, by immunocytochemistry using primary polyclonal rabbit antibody. A specific radioimmunoassay was used to detect endogenous concentration of ET in homogenized placental tissues. The endogenous concentration of ET-IR was significantly greater in amnion than in chorion and placenta (amnion 249 +/- 13 fmol/g; chorion 190 +/- 11 fmol/g; placenta 169 +/- 14 fmol/g; means +/- S.E.M.; n = 12; P less than 0.01). No significant difference was seen before or after the onset of labour. The detection of ET-IR in placenta, chorion and amnion suggests that the ETs may play a role in the paracrine control of human uterine function.

Amnion↗
PubMed2003

The effects of amniotic membrane on polymorphonuclear cells.

OBJECTIVE: To investigate the effects of fresh and preserved amniotic membrane on polymorphonuclear neutrophils (PMNs) so as to understand the anti-inflammatory mechanism of amniotic membrane transplantation. METHODS: Conditioned medium was collected 48 hours after fresh or preserved amnions were cultured in DMEM and 5% CO(2) at 37 degrees C. Then, polymorphonuclear cells were cultured in conditioned culture or DMEM. Fluorescent microscopy with 4',6-diamidino-2-phenylindole (DAPI) staining and cytometry were performed 6, 9, 12, and 15 hours later. RESULTS: Apoptotic neutrophils were found in each group at different time points. The percentage of apoptotic cells at 6, 9, 12, and 15 hours after culture in the fresh and preserved amnion groups and the control group was 17.3%, 24.4%, 29.8%, 37.1%, and 16.2%, 20.1%, 23.7%, 27.7%, and 10.2%, 13.7%, 21.1%, 26.4%, respectively (t test, P(1) < 0.01, P(2) < 0.01 and P(3) < 0.01). CONCLUSION: Amniotic membrane can accelerate apoptosis of polymorphonuclear neutrophils, reduce inflammation, and prevent ocular surface collagen from resolution, indicating that fresh amnion might have a stronger effect than preserved amnion.

Amnion↗
PubMed2006

[Comparison study between two types of vaginoplasty].

OBJECTIVE: To investigate the clinical therapeutic effects of two types of vaginoplasty. METHODS: From January 1996 to March 2005, 63 patients with the congenital absence of the vagina were treated by two types of vaginoplasty. Of the 63 patients, 37 underwent vaginoplasty using the amnion and 26 underwent an improved laparoscopic Vecchietti operation. The durations of the operation and hospitalization, as well as the blood loss were compared between the two types of vaginoplasty. The vaginal moulds were improved during the operations. RESULTS: According to the follow-up for 2 months to 4 years in the 35 patients. Compared with vaginoplasty using the amnion, vaginoplasty by an improved laparoscopic Vecchietti operation had advantages of significantly shorter surgical duration, shorter hospitalization, and less blood loss (P < 0.05). After the operations, the artificial vagina of all the 63 patients could hold a speculum and the mucosa appeared so soft and smooth with normal lubrication. The married patients were satisfied with the intercourse. However, after vaginoplasty using the amnion, an infection of the amnion occurred in 3 patients, scar contracture in 2 patients, one of whom underwent scar incision 13 months after operation with a success; but the other refuse to accept another operation. But the improved laparoscopic Vecchietti operation achieved a success in the patients without any infection or scar contracture, according to the 2 month-2.5 years follow-up. CONCLUSION: The improved laparoscopic Vecchietti operation is a preferred procedure of constructing a vagina for the patients suffering from the congenital absence of the vagina.

Adult↗
PubMed1987

A comparative study on in vitro invasion rates by melanoma cells in the human amniotic basement membrane model versus in vivo tumor nodule formation in C57BL6 mice.

Invasion by murine B16-F10 melanoma cells was studied using the human amniotic basement membrane (HABM) assay. B16-F10 cells were collected after a single passage through the amnion and grown to near confluency. The cycle of plating, passaging, collecting, and culturing B16-F10 cells was repeated five times. The invasion rate for B16-F10 cells remained relatively unchanged after six passages through the amnion. Injection of first-passage B16-F10 cells into C57BL6 mice resulted in 29 lung tumors per animal whereas sixth-passage cells resulted in 300+ lung tumors. While there exists no correlation of the number of cells penetrating the amnion with colonization number, lung colonization appears correlated with increased number of passages through the amnion.

Amnion↗
PubMed1979

Further evidence for the role of prolactin on human fetoplacental osmoregulation.

The addition of ovine prolactin (oPRL) to the fetal side of human term amnion in vitro is associated with a decrease in membrane permeability to tritiated water (THO). As the concentration of oPRL is increased from 2.5 to 20.0 micrograms per milliliter, permeability is progressively impaired. The addition of a specific prolactin receptor antibody completely abolished the effect of oPRL. Also, the addition of Ouabain abolished the effect of oPRL. When an osmolic gradient was created using Dextran-10 on the maternal side of the amnion, the bulk flow of water in control and 3PRL-treated membranes was not significantly different. These findings suggest that PRL acts predominantly on the diffusional flow rather than the bulk flow of water across amnion and that it is a transcellular transport. These studies also represent indirect evidence for the existence of prolactin receptor sites at the level of the amnion.

Amnion↗
PubMed1984

Protein synthesis but not DNA synthesis is required for tumor cell invasion in vitro.

The effects of protein and DNA synthesis inhibitors on in vitro tumor cell invasion was studied in a quantitative invasion system using human amnion. Highly invasive M5076 tumor cells were incubated on the basement membrane (BM) of the amnion in the presence of cycloheximide, aphidicolin or sodium butyrate. Tumor cell penetration through the entire thickness of the amnion was measured after 24 h. Protein synthesis in the M5076 cells was inhibited 95% within 1 h by 10(-6)M cycloheximide while DNA synthesis was unaffected. Treatment with cycloheximide reduced the number of spontaneously invading cells by 82% and by 66% in the presence of the chemoattractant, formylmethyl leucine-phenylalanine (FMLP), known to stimulate invasiveness. DNA synthesis in the M5076 tumor cells was selectively inhibited with aphidicolin (10 micrograms/ml). Although the DNA synthesis was greatly reduced, no significant effect on invasiveness with or without FMLP was observed. A less specific inhibitor of DNA synthesis, sodium butyrate (1 mM), arrested cell proliferation in the G1 phase of the cell cycle. When the butyrate-treated M5076 cells were tested in the invasion assay, a 50% increase in the number of invasive cells was observed. All three inhibitors were used in concentrations that did not affect cell viability or cell attachment to the amnion. These data indicate that protein synthesis but not DNA synthesis and not cell proliferation are required for tumor cells to invade native connective tissue barriers in vitro.

Amnion↗
PubMed1976

[Investigations on the occurrence of mycoplasma in the amniotic fluid (author's transl)].

In order to elucidate the pathogenetic importance of mycoplasma in amnionitis and in infectious syndrome of the amnion, the amniotic fluid of 185 healthy pregnant women was investigated for mycoplasma, bacteria and fungi. In no case mycoplasma were detected. This means that amniotic fluid is normally free of mycoplasma. If mycoplasma occurs in the amniotic fluid it has to be considered as a possible cause of amnionitis and infectious syndrome of the amnion.

Amnion↗