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PubMed1999

Amnion-derived cells express intercellular adhesion molecule-1: regulation by cytokines.

We have examined the expression of the intercellular adhesion molecule-1 (ICAM-1) mRNA in primary and established amnion-derived cell cultures and regulation of this expression by tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta. TNF-alpha (50 ng/ml) and IL-1beta (1.0 ng/ml) induced 18- and 11-fold increases respectively in expression of the ICAM-1 mRNA in WISH cells (an amnion epithelium-derived cell line). The increase was detectable within one hour of treatment and peaked by two hours. The protein synthesis inhibitor, cycloheximide (10 microg/ml) did not inhibit this induction. Increased levels of ICAM-1 protein were detected in the cells within 4 h after initiation of treatment with either cytokine. By 16 h of treatment with IL-1beta or TNF-alpha ICAM-1 reached 40 and 73 pg/microg cellular protein, representing 6- and 11-fold stimulations respectively. In primary amnion cells, basal expression of ICAM-1 mRNA was undetectable. However, TNF-alpha (50 ng/ml) induced ICAM-1 mRNA within two hours, peak expression being reached between four and eight hours after initiation of treatment. The present report demonstrates for the first time that amnion derived cells can express ICAM-1 and, further, that this expression is regulated by pro-inflammatory cytokines. This has implications for the amnion as a possible source for soluble ICAM-1, for this gene product as a marker for preterm labour, and for participation of the amnion, additional to its reported secretory role, in inflammatory processes of the fetal membranes.

Amnion↗
PubMed1990

Steroid, corticotrophin-releasing hormone, ACTH and prostaglandin interactions in the amnion and placenta of early pregnancy in man.

Corticotrophin-releasing hormone (CRH) is produced by both the placenta and fetal membranes at term in man, and CRH mRNA has been detected in human placental tissue. The synthesis of CRH and its control during early pregnancy, however, have not been established, and the role of CRH produced in the placenta and fetal membranes is not known. We examined whether amnion and placental tissue obtained between 12 and 15 weeks of gestation produced CRH in vitro, whether steroid modulation of output occurred and whether CRH affected prostaglandin (PG) output by the placenta and amnion. Immunoreactive (ir) CRH output by amnion (2.8 +/- 0.31 (S.E.M.) nmol/10(5) cells) was significantly (P less than 0.01) greater than that from placenta (1.76 +/- 0.21 nmol/10(5) cells). Output of ir-CRH decreased in the presence of progesterone, but increased in the presence of cortisol and dexamethasone. There was no significant effect of progesterone or ir-CRH output by placental cells; however, ir-CRH output was increased in the presence of dexamethasone and cortisol. There was no significant effect of corticosterone on ir-CRH output by either amnion or placental cells. Both ACTH and CRH stimulated the output of PGE2 and PGF2 alpha by amnion cells. In contrast, there was no significant effect of PGE2 output by placental cells maintained in the presence of either human CRH or ACTH. Output of PGF2 alpha by placental cells was increased in the presence of both CRH and ACTH. We conclude that both amnion and placental tissue produce CRH in early gestation, and that this output is modulated by steroids.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗
PubMed1997

The expression of parathyroid hormone-related protein mRNA and immunoreactive protein in human amnion and choriodecidua is increased at term compared with preterm gestation.

Parathyroid hormone-related protein (PTHrP) gene expression and/or immunoreactive protein have previously been identified in the uterus and intrauterine gestational tissues. The putative roles of PTHrP during pregnancy include vasodilatation, regulation of placental calcium transfer, uterine smooth muscle relaxation and normal fetal development. The aims of this study were 1) to determine the tissue-specific and temporal expression of PTHrP mRNA and immunoreactive protein in human gestational tissues collected at preterm and term; and 2) to determine the effect of labour on PTHrP expression by collecting these tissues from women undergoing elective caesarean section (before labour), intra-partum caesarean section during spontaneous-onset labour (during labour), and women with spontaneous labour and normal vaginal delivery (after labour). Total RNA and protein were extracted from placenta, amnion (over placenta and reflected) and choriodecidua for analysis by Northern blot (using a specific human PTHrP cDNA probe), and by N-terminal PTHrP RIA respectively. In amnion over placenta, reflected amnion and choriodecidua both PTHrP mRNA relative abundance and immunoreactive protein were significantly elevated at term compared with preterm (P < 0.01). At term, both PTHrP and its mRNA were significantly greater in amnion than in placenta and choriodecidua (P < 0.05). Also, both PTHrP and its mRNA were significantly elevated in amnion over placenta compared with reflected amnion (P < 0.05). The expression of PTHrP and its mRNA did not change in association with term labour or rupture of the fetal membranes, therefore this study provides no evidence for a specific PTHrP role in the onset and/or maintenance of term labour. However, the significant up-regulation of PTHrP mRNA and protein in the fetal membranes at term compared with preterm suggests an important role in late human pregnancy.

Amnion↗
PubMed1993

The amnion produces little of the prostaglandin E2 detected on the decidual side of human fetal membranes.

Cultured intact fetal membrane disks initially produced high levels of prostaglandin E2 (PGE2) on the fetal and maternal sides which declined during four days of culture. The transfer of a bolus of PGE2 from the fetal side to the maternal side of the membrane ranged from 1% to 3% after 24 hours of culture, and was a minimum over the period of 48-72 hours from the start of the incubation. To assess the handling of PGE2 synthesized by the amnion, 3H-arachidonic acid was incorporated into cultured amnion and into the amnion side of cultured intact fetal membrane disks. Labelled amnion released 3H-PGE2 on both sides of the tissue, whereas similarly labelled cultured intact fetal membrane only had detectable levels of 3H-PGE2 on the fetal side. It was calculated that no more than 9.7 +/- 1.4% of the PGE2 synthesised by the amnion crossed to the maternal side of the membrane without being metabolised during the transfer through the membrane. These results are consistent with similar indirect methods which suggested that PGE2 from the amnion may have only a limited role in human labor, and indicates the importance of using appropriate culture systems to investigate intra-uterine prostaglandin production.

Amnion↗
PubMed1999

Amnion allografts prepared in the Central Tissue Bank in Warsaw.

Applications of allogenic amnion grafts range from wound dressing of severe burns, dermabrasions and lower extremity ulcer treatments to plastic surgery, laryngology and ophthalmology. The aim of the present study was to elaborate the method of processing, preservation and sterilization of human amnion allografts prepared as wound dressing used mainly for burned patients. During the amniotic sac processing (after separation of chorion) special attention was paid to ensure that the epithelial side of amnion is placed directly on polyester net used as a support. After application on the wound, the epithelial side with the basement membrane is facing outwards; this will promote migration, attachment and spreading of the host cells encouraging epithelialization. Human amnion allografts were preserved by lyophilization or deep-freezing and subsequently radiation-sterilized with a dose of 35 kGy. It has been observed, however, that lyophilized irradiated allografts are resorbed within a few days, while frozen irradiated ones better adhere to wound and persist even 3 weeks after grafting, therefore, it has been decided to preserve amnion by deep-freezing. Since the beginning of 1998 over 400 preserved radiation-sterilized amnion allografts (with a total surface area over 40,000 cm2) have been prepared at the Central Tissue Bank in Warsaw and distributed to clinics and hospitals throughout the country.

Amnion↗
PubMed2002

Expression of angiogenic and neurotrophic factors in the human amnion and choriodecidua.

OBJECTIVE: Our objective was to identify the novel or differential expression of growth or development associated genes in the human gestational membranes that might play roles in pregnancy or in term or preterm parturition. STUDY DESIGN: Complementary DNA arrays were probed with [alpha(33)P]dCTP-labeled-complementary DNA that was prepared from the RNA of reflected amnion and choriodecidua that represent term not-in-labor, term spontaneous labor, and preterm labor with and without chorioamnionitis (n = 4 per group). Differential expression (term not-in-labor vs term spontaneous labor or preterm labor with chorioamnionitis vs preterm labor without chorioamnionitis) was evaluated by Wilcoxon tests. RESULTS: All 16 amnion samples expressed angiogenic factors (endothelin-2 and -3, vascular endothelial growth factor, and vascular endothelial growth factor-B) and neurotrophic factors (ephrin-A2, ephrin receptors-A2, -B1, -B3, -B4, and -B5, neuropilin-2, p75/nerve growth factor receptor and semaphorin-F). In both amnion and choriodecidua, the expression of vascular endothelial growth factor and the angiopoietin receptor, Tie-2, were greater with term spontaneous labor than with term not-in-labor (P <.05); increased VEGF receptor-2 (flk-1) expression was observed in term spontaneous labor choriodecidua (P <.05) but not amnion. Ephrin-A1 expression increased with term spontaneous labor in both tissues (P <.05). Semaphorin-F expression decreased with preterm labor with chorioamnionitis in choriodecidua (P <.05), although the trend was not significant in amnion (P =.1). CONCLUSION: Neurotrophic and angiogenic factor genes are expressed in amnion and choriodecidual membranes. Several of the genes exhibit differential expression with labor at term or in association with infection preterm, which suggests roles in or associated with these processes.

Amnion↗
PubMed2005

The application of deep-frozen and radiation-sterilized human amnion as a biological dressing to prevent prolonged air leakage in thoracic surgery.

Amnion allografts have been used in a number of clinical applications. However, no references of their use in thoracic surgery have been identified. Air leakage, is one of the most common complications in thoracic surgery, resulting from visceral pleura and lung injuries caused intraoperatively, in particular following rethoracotomy. Mechanical and manual sutures are also potential sites of air leakage. The aim of our study was to evaluate the usefulness of human amnion grafts in the treatment of air leakage following thoracic surgery. Deep-frozen, radiation-sterilized (35 kGy) human amnion grafts prepared in the Central Tissue Bank in Warsaw (Poland) were used. The amnion allografts were applied to 20 patients who had surgery: 11 thoracotomies and 9 rethoracotomies were performed (15 resections of 1-12 metastases, 3 lobectomies and 2 residual tumor resections). During lung ventilation the air leakage sites were covered by the amnion flap attached by moderate compression and stabilized by sutures. Air leakage and drainage were measured during the postoperative period. In 80% of the cases, no traces of air leakage was observed. The preliminary results suggest that the application of human amnion grafts may be a safe and effective method for preventing prolonged air leakage after thoracic surgery.

Adult↗
PubMedDate not supplied

[Use of fresh amnion in the treatment of corneal diseases].

The effects of fresh amnion on the course of morbid processes in corneal ulcers and burn disease of the anterior segment of the eye were investigated. The cornea was coated with amnion in 15 eyes with deep bacterial ulcers, in 10 eyes with herpetic ulcers of the cornea, and in 8 eyes with second and third-degree corneal and conjunctival burns. The amnion was obtained in cesarean section and fixed to the limbal conjunctiva with an uninterrupted suture all around. The next day after surgery the pain syndrome reduced, as did photophobia and blepharospasm in all the patients. In the group of patients with bacterial ulcers the cornea epithelialized on days 5-11, in those with herpetic ulcers on days 10-15, and in those with burns on days 8-12 after amnion coating. No cases with suppuration of the burnt surface when coated with the amnion were recorded. The amnion slid off the cornea on days 7-10 after it was layered on.

Amnion↗
PubMed1987

Human amnion membrane as a substratum for cultured peripheral and central nervous system neurons.

We report here on the use of human amnion membrane as a substratum for the culture of neuronal cells. Pieces of amnion membrane were bound to nitrocellulose paper as a supporting material, seeded with neurons, and cultured for 1-4 days. Neurons and neurites were visualized after fixation by immunoperoxidase staining using an anti-neurofilament monoclonal antibody. Neurons from embryonic chick ciliary and dorsal root ganglia and fetal rat hippocampus were cultured on either the basement membrane or stromal surface of the amnion membrane. Neurons adhered to both surfaces but extended neurites only on the basement membrane surface. Neurons survived and continued to grow neurites in serum-free medium for at least 4 days. When cultured for 4 days on the basement membrane surface in the presence of 10% fetal calf serum, the ciliary ganglion neurons survived but neurite growth was markedly inhibited, while dorsal root ganglion neurons survived, hypertrophied and grew an extensive network of neurites. Other experiments addressed the question whether the basement membrane surface had an ability to guide growing neurites. Amnion membranes were folded, frozen, cross-sectioned using a cryostat, and placed on the nitrocellulose to give irregular patterns of basement membrane juxtaposed to the collagenous stroma. Ciliary ganglion neurons after 4 days in culture had initiated and extended neurites in patterns which corresponded and were limited to those areas visualized by indirect immunofluorescence staining using anti-laminin antibodies. Thus, in vivo assembled human amnion basement membranes appear to contain signals that both promote and guide neuritic growth from previously axotomized embryonic peripheral and central nervous system neurons. The amnion membrane represents a novel tool for the culture of neuronal cells in vitro and potentially could be used as a neurite-promoting bridging material in vivo for regeneration studies.

Amnion↗
PubMedDate not supplied

[The participation of neurotransmitters in the regulation of the motor activity of the chick embryo amnion].

The authors' data on time-related changes in the contractile activity of non-innervated smooth muscle amnion of the chick embryo devoid of blood vessels and on involvement of the amniotic fluid neurotransmitters in regulation of the amnion motor activity have been summarized. A scheme of possible mechanism underlying regulation of the spontaneous rhythmic contractile activity of the amnion by two antagonistic neurotransmitters present in the amniotic fluid: serotonin and noradrenalin, which stimulate and inhibit the amnion motor activity, respectively. The results of experiments on successive block of the serotonin and adrenergic receptors by introduction of the corresponding antagonists in the chick embryo amniotic fluid speak in favor of the proposed scheme of humoral regulation of the amnion motor activity. A possible involvement of this humoral mechanism in the inhibitory effect of increased carbon dioxide content of the air on the amnion motor activity has been considered.

Amnion↗
PubMed1981

Compression-related defects from early amnion rupture: evidence for mechanical teratogenesis.

The features of 27 cases of limb/body wall deficiency (formerly termed cyllosomus and pleurosomus) were evaluated and the anomalies were interpreted as being band-related defects and/or compression-related defects. The latter included limb deficiency, body wall deficiency, neural tube defects, scoliosis, postural deformations, growth deficiency, and short umbilical cord. It is hypothesized that the single event of early amnion rupture can explain both the band-related defects and the compression-related defects. Experimental animal studies are in accord with this hypothesis; amnion puncture of rat fetuses during early gestation produces a comparable array of defects. The term amnion rupture sequence is suggested to describe the overall pattern of malformation that results from amnion rupture whether these defects are band related, compression related, or a combination of the two. There is considerable variation in the phenotype of amnion rupture sequence, with limb/body wall deficiency representing the more severe end of the spectrum. It is important to recognize and correctly diagnose amnion rupture sequence because it is usually a sporadic event.

Abdominal Muscles↗
PubMed1999

Amnion-chorion separation after 17 weeks' gestation.

OBJECTIVE: To evaluate the cause of and perinatal outcomes of amnion-chorion separation that is apparent sonographically after 17 weeks' gestation. METHODS: We searched our ultrasound database over 7 years for information on pregnant women who had live fetuses and complete separation between amnion and chorion that persisted beyond 17 weeks' gestation. For inclusion in the study, the women had to have amnion separated from chorion on at least three sides of the gestational sac. Medical records were reviewed for whether women had amniocenteses, results of the amniocenteses, and outcomes of the pregnancies. RESULTS: Of 15 pregnant women with live fetuses, ten had amniocenteses before identification of amnion-chorion separation and five did not. Three had fetuses with Down syndrome, two of whom had amnion-chorion separation evident before amniocentesis, and all three had other sonographic findings suggestive of aneuploidy. Three fetuses died. The other pregnancies were complicated by one or more adverse events, including two fetuses with growth restriction, five preterm deliveries, two with oligohydramnios, and one with abruptio placentae. Five infants were delivered at term and are alive and well. Overall, ten of 15 pregnancies resulted in live newborns, one of whom had Down syndrome. CONCLUSION: Complete amnion-chorion separation that persisted after 17 weeks' gestation is associated with a variety of adverse perinatal outcomes, including aneuploidy.

Extraembryonic Membranes↗
PubMed1981

Use of dry human and bovine amnion as a biological dressing.

A new method of drying and sterilizing the amnion extends the life of the product to more than nine months and enables it to be stored at room temperature. The bovine amnion is unique and the first of its kind, there being no comparable series in the literature, to our knowledge. Bovine amnion has a large surface area and can provide bulk supplies. The membrane as prepared has been tested in 12 rabbits experimentally and in 70 patients of different ages with a variety of surgical lesions, mainly burns. More than 150 applications of the dried amnion have been made as the sole dressing for these patients. The results indicate that amnion, both human and bovine, is quite safe and effective as a biological dressing. The dried material retains all the physical and biological characteristics of the fresh membrane.

Adolescent↗
PubMedDate not supplied

Sonographic determination of first trimester twin chorionicity and amnionicity.

OBJECTIVE: To determine the reliability of first trimester sonography in the detection of twin chorionicity and amnionicity. MATERIALS AND METHODS: Sets of twins (179) were evaluated sonographically in the first trimester. Chorionicity was determined by identifying the number of gestational sacs. The presence or absence of an amnion about each embryo/fetus was also recorded. The placentas for each case were evaluated to determine chorionicity and amnionicity. RESULTS: Of the 179 twin pairs, 158 were dichorionic/diamniotic; 19 were monochorionic/diamniotic, and two pairs were monochorionic/monoamniotic. Chorionicity and amnionicity were correctly determined in each case. CONCLUSION: The chorionicity of twin pregnancies can be determined from the time that the gestational sacs are visualized within the uterus. Transvaginal sonography can reliably determine the amnionicity of monochorionic twin pregnancies after 7 weeks, menstrual age.

Amnion↗
PubMed1986

Evidence that expression of c-fos protein in amnion cells is regulated by external signals.

While c-fos expression is normally very low in certain cell types but transiently inducible by growth factors (e.g. in fibroblasts), other cells (e.g. amnion cells) exhibit an apparently constitutive expression in vivo. Here we show that in primary amnion cells c-fos protein expression rapidly drops to undetectable levels following plating in normal growth medium. However, c-fos expression is inducible by, and maintained at high levels in the presence of, dialyzed placenta- or embryo-conditioned medium. These observations suggest that c-fos expression in primary amnion cells is regulated by placenta- and embryo-derived factor(s), providing further evidence for the hypothesis that transcription of the c-fos gene may generally be controlled by external signals. We also show that proliferation of primary amnion cells is not dependent on a high c-fos expression, suggesting that the function of c-fos is more likely to be associated with other cellular functions in the differentiated amnion cell.

Amnion↗
PubMed1992

Changes in annexin (lipocortin) content in human amnion and chorion at parturition.

Arachidonic acid is mobilized from fetal membrane phospholipids at parturition leading to increased production of oxytocic prostaglandins which may initiate or maintain myometrial contractions. Phospholipid mobilization requires activation of phospholipase A2 or C, both of which require calcium for activity. The annexins (lipocortins) are a superfamily of proteins which bind to calcium and phospholipids and thereby may alter phospholipase activity through two mechanisms: modulation of intracellular free Ca2+ concentrations or regulation of the accessibility of phospholipids to hydrolyzing enzymes. Using Western immunoblotting with monospecific polyclonal antibodies, annexins I-VI were identified in human amnion and chorion/decidua at term in tissues obtained from patients in labor or not in labor. Each annexin was present in two distinct pools: a pool which only associated with the membrane in the presence of calcium (calcium-dependent pool) and a calcium-independent pool that remained membrane bound in the presence of calcium chelators. Annexin I was present as two species, resolving at 36 kDa and 68 kDa. The total concentration of annexin I in both amnion and chorion/decidua was significantly decreased with labor, while the total concentration of annexin V in chorion significantly increased with labor. The size of individual pools of annexins also changed with labor: the calcium-dependent pool of annexins I and II in both amnion and chorion significantly decreased; the calcium-dependent pool of annexin V increased in chorion; and calcium-independent pools of annexin I in amnion and annexins I, II, and V in chorion significantly decreased with labor. The decrease in total annexin I concentration with labor in amnion reflects a substantial decrease (80-90%) in the pool tightly bound to the membrane in a calcium-independent manner. This striking change distinguishes annexin I as a potential candidate inhibitor which is specifically downregulated at parturition, potentially leading to increased access of phospholipases to substrate phospholipids and increased prostaglandin production at labor.

Amnion↗
PubMed1992

Amnion cell biosynthesis of interleukin-8: regulation by inflammatory cytokines.

The cellular constituents of the placenta are important participants in the recruitment and trafficking of inflammatory cells within the placenta. In infection-induced labor, gestational tissues synthesize and release a variety of inflammatory cytokines whose effects include increased prostaglandin biosynthesis and the initiation of uterine contractions. Interleukin-8 (IL-8), a potent neutrophil chemoattractant, has been recently described as being elevated in the amniotic fluid of mothers with chorioamnionitis. We investigated the biosynthesis of IL-8 by human amnion cells and its regulation by other inflammatory cytokines. Cultured amnion cells obtained from normal term placentae were found to produce IL-8 in response to pathophysiologic concentrations of interleukin 1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF-alpha). Treatment of amnion cells stimulated by IL-1 beta with cycloheximide resulted in increased IL-8 production, while incubation of IL-1 beta treated amnion cells with actinomycin D resulted in a concentration-dependent decrease in detectable amounts of IL-8. Northern blot analysis of cultured amnion cells stimulated with IL-1 beta demonstrated a rapid increase in IL-8 mRNA which peaked at 2-4 hr. These in vitro results suggest inflammation of gestational tissues in vivo may result in locally produced IL-8 and, in association with other inflammatory mediators, may be important in the pathophysiology of infection-induced labor.

Amnion↗
PubMed2001

Role of motility in embryonic development I: Embryo movements and amnion contractions in the chick and the influence of illumination.

This study provides a quantitative analysis of the active movements of the chick embryo and of the contractions of the amnion over the entire developmental period of 21 days. Four types of embryo movements are distinguished. The motor activity of the embryo shows two characteristic peaks, with maximum contraction frequencies on the 12th and on the 16th day. In contrast, the amnion activity is higher at earlier stages and decreases as the body activity increases. The amnion activity is largely independent of the body activity. Illumination has a strong influence on embryo movements. It is shown that increases of light intensity affect the patterns of activity of both the embryo and the amnion. While the effect of light on the embryo can be interpreted as being transmitted via the optic system, the mechanism of the amniotic response is unclear. The results suggest that the amnion itself may be sensitive to light. J. Exp. Zool. (Mol. Dev. Evol.) 291:186-194, 2001.

Amnion↗