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PubMed2001

The amniotic fluid as a source of cells for fetal tissue engineering.

PURPOSE: This study was aimed at determining whether fetal tissue constructs can be engineered from cells normally found in the amniotic fluid. METHODS: A subpopulation of morphologically distinct cells was isolated mechanically from the amniotic fluid of pregnant ewes (n = 5) and expanded selectively. Its lineage was determined by immunofluorescent staining against multiple intermediate filaments and surface antigens. Proliferation rates were determined by both oxidation and total DNA assays and compared with immunocytochemically identical adult and fetal sheep cells. Statistical analysis was by analysis of variance for repeated measures (ANOVA). After expansion, the amniocytes were seeded onto a polyglycolic acid polymer/poly-4-hydroxybutyrate scaffold. The resulting construct was analyzed by both optical and scanning electron microscopy. RESULTS: The immunocytochemical profile of expanded amniocytes was consistent with a mesenchymal, fibroblast/myofibroblast cell lineage. These cells proliferated significantly faster than comparable fetal and adult cells in culture. Amniocyte construct analysis showed dense, confluent layers of cells firmly attached to the scaffold, with no evidence of cell death. CONCLUSIONS: (1) Subpopulations of fetal mesenchymal cells can be isolated consistently from the amniotic fluid. (2) Mesenchymal amniocytes proliferate more rapidly in vitro than comparable fetal and adult cells. (3) Mesenchymal amniocytes attach firmly to polyglycolic acid polymer. The amniotic fluid can be a reliable and practical source of cells for the engineering of select fetal tissue constructs.

Amniotic Fluid↗
PubMedDate not supplied

Amniotic fluid interleukin-6 and the risk of early-onset sepsis among preterm infants.

BACKGROUND: High concentrations of interleukin-6 (IL-6) have been demonstrated in amniotic fluid (AF) from women with intra-amniotic infection. Recent studies have reported that IL-6 levels in AF were related to an increase in neonatal morbidity; moreover, higher IL-6 plasma levels have been observed in neonates with sepsis. METHODS: A cohort study was carried out at the National Institute of Perinatology in Mexico City. Inclusion criteria were the following: 1) preterm singleton pregnancy; 2) intact membranes at time of enrollment, and 3) written informed consent. Women with other complications of pregnancy were excluded. Newborn sepsis during the first 72 h was defined as early-onset sepsis. Amniotic fluid was obtained at the moment of delivery. Amniotic fluid IL-6 (AF IL-6) was determined by enzyme-linked immunoassays. RESULTS: Ninety-three women met the criteria for enrollment in the study and 31 (33%) of their newborns had early-onset neonatal sepsis. The mean AF IL-6 in mothers of septic newborns was 5779 +/- 2804 pg/ml compared to 729 +/- 382 pg/ml in mothers with non-infected neonates (p < 0.001). AF IL-6 concentrations higher than 1250 pg/ml were significantly associated with early-onset sepsis (OR 33.3; 95% CI 9.4-117.3) (p < 0.001). Gestational age under 32 weeks was also associated with neonatal sepsis (OR 2.56; 95% CI 1.2-9) (p = 0.002). Women whose infants developed neonatal sepsis had a higher frequency of clinical chorioamnionitis (p = 0.02). CONCLUSIONS: IL-6 determination in AF may be a useful indicator to identify neonates with higher risk of in utero bacterial infection.

Age of Onset↗
PubMed2004

Amniotic fluid concentrations of collagenase-1 and collagenase-3 are increased in polyhydramnios.

OBJECTIVE: Polyhydramnios places increased tension on the fetal membranes and should conceptually be associated with an increased need for tissue remodeling to prevent premature rupture of membranes (PROM). Herein we use polyhydramnios as a model to study the association of the collagenase class of matrix metalloproteinases (MMPs) with tissue remodeling during pregnancy. METHODS: Amniotic fluids were collected by transabdominal amniocentesis from women with polyhydramnios and from women with normal amniotic fluid volume at second trimester, third trimester and at term. Fluids were assayed for MMP-1 and MMP-13 (collagenase-1 and 3) using ELISA. The concentrations in polyhydramnios were compared with subjects undergoing amniocentesis with normal amniotic fluid volume. Statistical comparisons were made using Scheffe's method for correction in multiple means comparison. P of <0.05 was considered statistically significant. RESULTS: MMP-1 and MMP-13 are present in the amniotic fluid at all stages of pregnancy. The concentrations of these MMPs were significantly elevated in the polyhydramnios samples independent of gestational age. CONCLUSION: Polyhydramnios is associated with increased amniotic fluid concentrations of MMP-1 and MMP-13 suggesting their role in tissue remodeling.

Adult↗
PubMed1992

Nutrient levels in amniotic fluid from women with normal and neural tube defect pregnancies.

We analyzed nutrient levels in amniotic fluid obtained during the second trimester of normal, uncomplicated pregnancies from 221 women who delivered apparently healthy infants and from 8 with neural tube defect (NTD) pregnancies. Folate was measured by microbiological assay, vitamin B12 by a radiobinding method, and zinc, copper and iron by atomic absorption spectrophotometry. We found that the mean amniotic fluid nutrient levels of normal pregnancies were 24.7 nmol/l for folate, 600 pmol/l for vitamin B12, and 1.7, 1.9, and 9.0 mumol/l for zinc, copper and iron, respectively. Amniotic fluid folate, zinc, copper and iron levels of NTD pregnancies were similar to those found during normal pregnancy, however, vitamin B12 levels were markedly lower than those of normal pregnancies.

Adolescent↗
PubMed1993

[Cortisol in amniotic fluid during PGE2-induced labor].

Concentrations of cortisol in amniotic fluid had been estimated during induced deliveries by prostaglandin E2 (group PG) and induced ones by oxytocin (control group OX). Cortisol concentration in amniotic fluid was much higher in group PG. The mode of action of prostaglandin on fetal lung had been analyzed. Our results prefer prostaglandin as drug of choice for induction of labour.

Amniotic Fluid↗
PubMed2005

The detection of CMV in amniotic fluid and cervicovaginal smear samples by real-time PCR assay in prenatal diagnosis.

OBJECTIVE: There is no specific antiviral therapy or a vaccine, which could be safely administered to the pregnant women with primary human cytomegalovirus (CMV) infection. Therefore, prenatal diagnosis has a critical role in the management of pregnancy, complicated by this disease. In this study, we investigated the prevalence and clinical consequences of human CMV infection from cervicovaginal smear and amniotic fluid samples of pregnant women by using real-time polymerase chain reaction (RT-PCR) assay, in one of the Obstetrics and Gynecology outpatient clinics of Turkey. The identification of reliable prognostic markers of fetal disease remains the main purpose and a major challenge on this issue. METHODS: Two hundred and six samples, of which 135 were cervicovaginal smear and 71 were amniotic fluid, were enrolled in the study. The DNAs of the samples were extracted by using Roche Diagnostic (Roche, Germany) kit and amplifications of these DNAs were studied by using Light-Cycler system (Roche Germany) as being quantitative. Anti-CMV IgM antibodies in the samples were studied by both MEIA (Imx system, Abbot Laboratories, USA) and a commercial ELISA kit (Radim SPA, Italy) while anti-CMV IgG antibodies were studied by MEIA (Axsym system, Abbot Laboratories, USA). RESULTS: Human CMV DNA was found to be positive in 1.5% (2 in 135) of cervicovaginal smear and 1.4% (1 in 71) of amniotic fluid samples by RT-PCR. IgM and IgG were found to be negative in all of the cervicovaginal smear samples by both MEIA and ELISA, while IgG antibody was found to be positive in only one of the amniotic fluid samples by MEIA. CONCLUSION: With RT-PCR assay, we have found the prevalence of human CMV in pregnant women similar to epidemiologic reports, which have been described earlier. Whereas the fetus with positive amniotic fluid in favor of human CMV had an intrauterine growth restriction resulted in intrauterine exitus, no symptoms were observed in the infants of the other two pregnant women with positive RT-PCR results. The fact that the clinical consequence of the newborn whose amniotic fluid evaluation revealed human CMV infection by RT-PCR made us think that this molecular diagnosis method may be a reliable assay in prenatal diagnosis of this pathogen.

Amniotic Fluid↗
PubMed2001

Fetal urine and amniotic fluid in monochorionic twins with twin-twin transfusion syndrome: simulations of therapy.

OBJECTIVE: Therapy for twin-twin transfusion syndrome includes amnioreduction, septostomy, and laser ablation, but there is no accepted standard of care. Mechanisms for the reported improvement in survival are incompletely understood. We sought to model the mechanisms and predict the response of varying severities of twin-twin transfusion syndrome and placental angioarchitectures to current therapies to determine optimal clinical interventions. STUDY DESIGN: We used our mathematic model of twin-twin transfusion syndrome that predicts fetal and amniotic fluid abnormalities that are related to the placental angioarchitecture. Amnioreduction was modeled as reduction in amniotic fluid volume; septostomy was modeled as the merging of donor and recipient amniotic fluid, and laser ablation was modeled as the cessation of all placental anastomotic blood flows. RESULTS: Amnioreduction reduces amniotic fluid pressure, which improves transplacental fluid flow from mother to fetus and increases both donor and recipient blood volume. However, net arteriovenous transfusion increases, because of increased donor arterial pressure, which negates, in part, the benefit of amnioreduction. Septostomy allows amniotic fluid to be swallowed by the donor, with minimal effects on donor growth and blood volume. Laser ablation eliminates anastomotic exchange of blood and reduces discordant fetal growth. CONCLUSION: Amnioreduction may be effective in milder twin-twin transfusion syndrome pregnancies but ineffective in severe cases. Septostomy is unlikely to offer significant therapeutic efficacy. Laser ablation is equally effective in mild and severe twin-twin transfusion syndrome but has a higher spontaneous abortion rate than amnioreduction. The model indicates improved outcomes with the use of amnioreduction in mild twin-twin transfusion syndrome cases and with laser ablation in severe cases.

Amniocentesis↗
PubMedDate not supplied

[Measurement of fetoplacental blood flow (resistance index) during labor with meconium stained amniotic fluid using the doppler technic].

The blood flow in the fetoplacental circulation (umbilical artery) was examined by the Doppler technique in 42 labouring women with the meconium stained amniotic fluid. Ultrasound examination in the phase of cervical dilatation of 2-5 and 6-9 cm by the Doppler technique before and during contraction was carried out in each woman. Flow velocity wave-forms (FVW) of the umbilical artery were analysed by using the resistance index (RI). The ultrasonic apparatus Kranzbuhler ADR 8130 was used for measuring. The control group consisted of 54 labouring women with a clean, milky amniotic fluid. All resistance index (RI) mean values with the meconium amniotic fluid. All resistance showed a statistically significant decrease when compared to the group with the milky amniotic fluid (T = 5.91 p less than 0.01; t = 4.57 p less than 0.01; t = 5.73 p less than 0.01 t = 3.19 p less than 0.01). They probably are an indicator of the compensatory mechanism of the fetus.

Amniotic Fluid↗
PubMed1978

Organic acids in amniotic fluid.

The organic acids of human amniotic fluid were studied by gas chromatography and mass spectrometry. About 30 acids were identified. A "normal" metabolic profile of acids was established which can provide the basis for the identification of abnormal patterns.

Amniotic Fluid↗
PubMed1997

Midtrimester amniotic fluid tumor necrosis factor-alpha does not predict small-for-gestational-age infants.

PROBLEM: To evaluate the independent ability of midtrimester amniotic fluid tumor necrosis factor-alpha (TNF-alpha) in the prediction of small-for-gestational-age (SGA) infants. METHOD OF STUDY: In this case-control study, patients delivering a SGA infant were matched with controls based on GA at delivery, maternal age, race, and parity. Patients with immune disease, chronic hypertension, diabetes, asthma, congenital hearts disease, multiple gestation, and fetal anomalies were excluded. Amniotic fluid samples were immunoassayed for TNF-alpha. Potential confounding variables evaluated were maternal serum alpha-fetoprotein level, smoking history, pregnancy induced hypertension, and neonatal gender. Statistical analysis included Fisher's exact test and ANOVA after log transformation with P < 0.05 considered significant. RESULTS: Eighteen patients delivered SGA neonates and were matched with 41 controls. No significant differences were identified in the confounding variables between patients with SGA neonates and controls. Amniotic fluid TNF-alpha levels were not significantly different between patients subsequently delivering SGA neonates and controls [median 7.63 (range 0.25-16.1) pg/mL versus 9.39 (0.25-66.9) pg/mL, P = 0.8]. CONCLUSIONS: Midtrimester amniotic fluid TNF-alpha levels are not predictive of SGA neonates when compared with controls matched for gestational age at delivery.

Amniotic Fluid↗
PubMed1979

The iodine content of amniotic fluid and placental transfer of iodinated drugs.

Determinations of total iodine content, levels of thyroxine (T4) and triiodothyronine (T3), and protein concentrations were made on amniotic fluid. A total of 218 samples obtained at various stages of pregnancy, from both normal and pathological pregnancies, were studied. Normal values for the iodine and hormone concentrations are presented. Several amniotic fluid samples showed greatly elevated iodine levels, thought to be the result of maternal iodine intake. This study demonstrated that such elevated levels can be produced by urography with an iodinated medium, by thyroid extract therapy, and by vaginal therapy with an iodinated agent. Levels of T4 and T3 in amniotic fluid decreased slightly from the second to the third trimester and were unrelated to the total iodine levels. Despite reports in the literature of fetal hypothyroidism produced by amniography with a fat-soluble medium, no thyroid defect was observed in this series in the presence of elevated amniotic fluid iodine levels or when a water-soluble iodinated medium was used for amniography.

Amniotic Fluid↗
PubMed1991

Isolation of amniotic fluid proteins of non-maternal serum origin by negative immuno-affinity chromatography.

Although several proteins of non-maternal serum origin are known to exist in amniotic fluid, the predominance of serum proteins has hampered the isolation of all but a very few. This report describes the removal of large amounts contaminating serum proteins from amniotic fluid with anti-human serum antibody coupled to a solid matrix, leaving behind small amounts of proteins of interest, about 3% of the total amniotic fluid proteins. An antibody raised against the non-adsorbed proteins and used in immunoelectrophoretic methods permitted detection of more, probably unknown, proteins in amniotic fluid of potential biological and diagnostic interest.

Amniocentesis↗
PubMed2000

Elevations of group II phospholipase A2 concentrations in serum and amniotic fluid in association with preterm labor.

OBJECTIVE: The purpose of this study was to determine whether the elevation of secretory group II phospholipase A(2) concentration in the serum and amniotic fluid in preterm labor is associated with intrauterine inflammation. STUDY DESIGN: Serum and amniotic fluid were collected from women with preterm delivery (<37 weeks' gestation; n = 38) and term delivery (n = 20). Phospholipase activity was measured with a highly sensitive system that was based on reverse-phase high-performance liquid chromatographic separation of 9-anthracenylmethyl derivatives of fatty acids released by phospholipase A(2). The concentrations of immunoreactive isozymes (group I or II) of secretory phospholipase A(2) were assayed with a radioimmunoassay kit with a monoclonal antibody against human pancreatic phospholipase A(2) and splenic IIA phospholipase A(2). Localization of immunoreactive group II phospholipase A(2) in the amniotic membrane was determined by immunostaining visualized with the Vectastain ABC (Vector Laboratories, Inc, Burlingame, Calif) method. RESULTS: Enzymatic activities of phospholipase A(2) in the serum and amniotic fluid specimens obtained from patients in preterm labor with chorioamnionitis were significantly higher than those in specimens from patients in term labor. Significant elevations of phospholipase A(2) activities were observed in patients with preterm labor without histologically evident chorioamnionitis. The activity of phospholipase A(2) was clearly correlated with the concentration of the immunoreactive group II phospholipase A(2). Group II phospholipase A(2) was localized in amniotic cells obtained from patients with a pathologically determined diagnosis of chorioamnionitis. The predictive value for chorioamnionitis of the group II phospholipase A(2) concentration was relatively higher than the predictive values of the concentrations of C-reactive protein and interleukins 6 and 8. CONCLUSION: Significant elevations of group II phospholipase A(2) concentrations were detected in the serum and amniotic fluid of women with preterm labor. Group II phospholipase A(2) concentration may be a useful indicator for preterm labor, and phospholipid metabolism is certainly activated both in preterm labor and in apparent inflammatory diseases.

Amniotic Fluid↗
PubMed2005

Amniotic fluid: not just fetal urine anymore.

Amniotic fluid (AF) is a complex substance essential to fetal well-being. This article reviews recent discoveries and the current understanding of the origin and circulation of AF and its nutritive, protective, and diagnostic functions. Future directions for AF research are also discussed.

Amniotic Fluid↗
PubMed1989

Normal amniotic fluid volume changes throughout pregnancy.

The purpose of this study was to provide a quantitative characterization of the changes in amniotic fluid volume that occur throughout gestation. From 705 published amniotic volumes for normal pregnancies, we found that after log transformation, amniotic fluid volume had a uniform variability over 8 to 43 weeks' gestation. Thus the 95% confidence interval covered the range of 1/2.57 to 2.57 times the mean volume at any given gestational age. Contrary to expected trends, mean amniotic fluid volume did not change significantly between 22 and 39 weeks and averaged 777 ml, with the 95% confidence interval ranging from 302 to 1997 ml. The data are summarized in nomograms covering 8 to 43 weeks' gestation.

Amniotic Fluid↗