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Methods in Enzymology 2020Extracellular vesicles (EVs), specifically exosomes of 50-150nm, have emerged as important communication channels between cells and tissues and can be isolated from...
Extracellular vesicles (EVs), specifically exosomes of 50-150nm, have emerged as important communication channels between cells and tissues and can be isolated from multiple biofluids including blood, urine and amniotic fluid. No standardized approach for exosome isolation from these biofluids has been established. This chapter outlines an optimized approach for isolating exosomes from human amniotic fluid samples. Like plasma, amniotic fluid contains many protein and cellular contaminants that requires multiple steps for cleanup. Therefore, to ensure samples contain minimal contaminants, including larger EVs, we also outline multiple methods for characterization of isolated exosomes for size, morphology and protein markers.
Topics: Amniotic Fluid; Biomarkers; Exosomes; Extracellular Vesicles; Humans
PubMed: 33565971
DOI: 10.1016/bs.mie.2020.07.006 -
Journal of Extracellular Vesicles May 2022Amniotic fluid surrounding the developing fetus is a complex biological fluid rich in metabolically active bio-factors. The presence of extracellular vesicles (EVs) in...
Amniotic fluid surrounding the developing fetus is a complex biological fluid rich in metabolically active bio-factors. The presence of extracellular vesicles (EVs) in amniotic fluid has been mainly related to foetal urine. We here characterized EVs from term amniotic fluid in terms of surface marker expression using different orthogonal techniques. EVs appeared to be a heterogeneous population expressing markers of renal, placental, epithelial and stem cells. Moreover, we compared amniotic fluid EVs from normal pregnancies with those of preeclampsia, a hypertensive disorder affecting up to 8% of pregnancies worldwide. An increase of CD105 (endoglin) expressing EVs was observed in preeclamptic amniotic fluid by bead-based cytofluorimetric analysis, and further confirmed using a chip-based analysis. HLA-G, a typical placental marker, was not co-expressed by the majority of CD105 EVs, in analogy with amniotic fluid stromal cell derived-EVs. At a functional level, preeclampsia-derived EVs, but not normal pregnancy EVs, showed an antiangiogenic effect, possibly due to the decoy effect of endoglin. Our results provide a characterization of term amniotic fluid-EVs, supporting their origin from foetal and placental cells. In preeclampsia, the observed antiangiogenic characteristics of amniotic fluid-EVs may reflect the hypoxic and antiangiogenic microenvironment and could possibly impact on the developing fetus or on the surrounding foetal membranes.
Topics: Amniotic Fluid; Biomarkers; Endoglin; Extracellular Vesicles; Female; Humans; Phenotype; Placenta; Pre-Eclampsia; Pregnancy
PubMed: 35582873
DOI: 10.1002/jev2.12217 -
Journal of Perinatal Medicine Sep 2023This study was conducted to determine whether bacteria, fungi, or archaea are detected in the amniotic fluid of patients who underwent midtrimester amniocentesis for...
OBJECTIVES
This study was conducted to determine whether bacteria, fungi, or archaea are detected in the amniotic fluid of patients who underwent midtrimester amniocentesis for clinical indications.
METHODS
Amniotic fluid samples from 692 pregnancies were tested by using a combination of culture and end-point polymerase chain reaction (PCR) techniques. Intra-amniotic inflammation was defined as an interleukin-6 concentration >2,935 pg/mL.
RESULTS
Microorganisms were detected in 0.3% (2/692) of cases based on cultivation, 1.73% (12/692) based on broad-range end-point PCR, and 2% (14/692) based on the combination of both methods. However, most (13/14) of these cases did not have evidence of intra-amniotic inflammation and delivered at term. Therefore, a positive culture or end-point PCR in most patients appears to have no apparent clinical significance.
CONCLUSIONS
Amniotic fluid in the midtrimester of pregnancy generally does not contain bacteria, fungi, or archaea. Interpretation of amniotic fluid culture and molecular microbiologic results is aided by the assessment of the inflammatory state of the amniotic cavity. The presence of microorganisms, as determined by culture or a microbial signal in the absence of intra-amniotic inflammation, appears to be a benign condition.
Topics: Pregnancy; Female; Humans; Amniotic Fluid; Pregnancy Trimester, Second; Chorioamnionitis; Archaea; Retrospective Studies; Bacteria; Inflammation; Fungi
PubMed: 37194083
DOI: 10.1515/jpm-2022-0604 -
European Journal of Pediatrics Dec 2019
Neonatologists and non-vigorous newborns with meconium-stained amniotic fluid (MSAF) in the delivery room: time for hands off? : Comment on: Kumar A, Kumar P, Basu S. "Endotracheal Suctioning for Prevention of Meconium Aspiration Syndrome: A Randomized Controlled Trial." European Journal of...
Topics: Humans; Infant, Newborn; Pregnancy; Amniotic Fluid; Delivery Rooms; Meconium; Meconium Aspiration Syndrome; Neonatologists; Pediatrics; Female
PubMed: 31667571
DOI: 10.1007/s00431-019-03501-w -
MicroRNA (Shariqah, United Arab... 2020The study aimed to provide an overall view of current data considering the presence of microRNAs in amniotic fluid. (Review)
Review
AIM
The study aimed to provide an overall view of current data considering the presence of microRNAs in amniotic fluid.
METHODS
The available literature in MEDLINE, regarding the role of the amniotic fluid in pregnancy and fetal development, was searched for related articles including terms such as "microRNA", "Amniotic fluid", "Adverse outcome" and others.
RESULTS
The amniotic fluid has an undoubtedly significant part in fetal nutrition, with a protecting and thermoregulatory role alongside. MicroRNAs have proven to be highly expressed during pregnancy in many body liquids including amniotic fluid and are transferred between cells loaded in exosomes, while they are also implicated in many processes during fetal development and could be potential biomarkers for early prediction of adverse outcomes.
CONCLUSION
Current knowledge reveals that amniotic fluid microRNAs participate in many developmental and physiological processes of pregnancy including proliferation of fibroblasts, fetal development, angiogenesis, cardioprotection, activation of signaling pathways, differentiation and cell motility, while the expression profile of specific microRNAs has a potential prognostic role in the prediction of Down syndrome, congenital hydronephrosis and kidney fibrosis.
Topics: Amniotic Fluid; Body Fluids; Cell Differentiation; Female; Fetal Development; Genetic Markers; Humans; MicroRNAs; Pregnancy; Signal Transduction
PubMed: 30887932
DOI: 10.2174/2211536608666190318105140 -
PloS One 2020Amniotic fluid is clinically accessible via amniocentesis and its protein composition may correspond to birth timing. Early changes in the amniotic fluid proteome could...
BACKGROUND
Amniotic fluid is clinically accessible via amniocentesis and its protein composition may correspond to birth timing. Early changes in the amniotic fluid proteome could therefore be associated with the subsequent development of spontaneous preterm delivery.
OBJECTIVE
The main objective of this study was to perform unbiased proteomics analysis of the association between mid-trimester amniotic fluid proteome and spontaneous preterm delivery and gestational duration, respectively. A secondary objective was to validate and replicate the findings by enzyme-linked immunosorbent assay using a second independent cohort.
METHODS
Women undergoing a mid-trimester genetic amniocentesis at Sahlgrenska University Hospital/Östra between September 2008 and September 2011 were enrolled in this study, designed in three analytical stages; 1) an unbiased proteomic discovery phase using LC-MS analysis of 22 women with subsequent spontaneous preterm delivery (cases) and 37 women who delivered at term (controls), 2) a validation phase of proteins of interest identified in stage 1, and 3) a replication phase of the proteins that passed validation using a second independent cohort consisting of 20 cases and 40 matched controls.
RESULTS
Nine proteins were nominally significantly associated with both spontaneous preterm delivery and gestational duration, after adjustment for gestational age at sampling, but none of the proteins were significant after correction for multiple testing. Several of these proteins have previously been described as being associated with spontaneous PTD etiology and six of them were thus validated using enzyme linked immunosorbent assay. Two of the proteins passed validation; Neutrophil gelatinase-associated lipocalin and plasminogen activator inhibitor 1, but the results could not be replicated in a second cohort.
CONCLUSIONS
Neutrophil gelatinase-associated lipocalin and Plasminogen activator inhibitor 1 are potential biomarkers of spontaneous preterm delivery and gestational duration but the findings could not be replicated. The negative findings are supported by the fact that none of the nine proteins from the exploratory phase were significant after correction for multiple testing.
Topics: Adult; Amniocentesis; Amniotic Fluid; Cohort Studies; Female; Gestational Age; Humans; Male; Pregnancy; Pregnancy Trimester, Second; Premature Birth; Proteome
PubMed: 32379834
DOI: 10.1371/journal.pone.0232553 -
Neurotoxicology and Teratology 2023Prenatal stress adversely affects offspring development, with fetal cortisol (CORT) exposure being a primary hypothesized mechanism for stress-induced developmental...
Prenatal stress adversely affects offspring development, with fetal cortisol (CORT) exposure being a primary hypothesized mechanism for stress-induced developmental deficits. Fetal CORT exposure can be assessed via measurements in amniotic fluid. However, in humans, amniocentesis is typically only performed for clinical reasons such as karyotyping; thus, amniotic fluid CORT cannot be obtained from a random sample. To test the hypothesis that fetal CORT exposure predicts neonatal and infant development in healthy primates, we measured amniotic fluid CORT in N = 18 healthy rhesus macaque (Macaca mulatta) dams (50:50 female:male infants) between 80 and 124 days gestation (mean ± SEM = 98.3 ± 2.9 days out of 165 days gestational length; i.e., second trimester). Maternal hair cortisol concentrations (HCCs) were assessed throughout pregnancy and lactation. Offspring were assessed for physical growth, neurological development, cognitive development, and HCCs across postnatal days 30-180. Controlling for gestational age at amniocentesis, higher amniotic fluid CORT significantly predicted slower infant growth rate (g/day) in the first 30 days (β = -0.19; R = 0.71, p = .008), poorer sensorimotor scores on the day 30 neonatal assessment (β = -0.28; R = 0.76, p = .015), and longer time to complete training (β = 0.48; R = 0.54, p = .026), but better performance (β = 0.91; R = 0.60, p = .011) on a discrimination cognitive task at 120-180 days. Amniotic fluid CORT was not associated with maternal or infant HCCs. Although these results are correlative, they raise the intriguing possibility that fetal CORT exposure in non-stress-exposed primates, as measured by amniotic fluid CORT, programs multiple aspects of neonatal and infant development. On the other hand, amniotic fluid CORT may not relate to chronic CORT levels in either mothers or infants when assessed by hair sampling.
Topics: Pregnancy; Infant, Newborn; Infant; Animals; Child; Humans; Female; Male; Amniotic Fluid; Macaca mulatta; Hydrocortisone; Child Development; Amniocentesis
PubMed: 37890675
DOI: 10.1016/j.ntt.2023.107308 -
The Journal of Maternal-fetal &... Dec 2023The aim of this review is to evaluate the relationship between the use of non-steroidal anti-inflammatory drugs (NSAIDs) during last trimesters of the pregnancy and the... (Review)
Review
OBJECTIVE
The aim of this review is to evaluate the relationship between the use of non-steroidal anti-inflammatory drugs (NSAIDs) during last trimesters of the pregnancy and the reduction of amniotic fluid.
METHODS
Electronic databases were searched (PubMed, Medline, and Scopus). Selection criteria included studies reporting the relationship between oligohydramnios and use of NSAID during pregnancy. We analyzed the median age of women, weeks of pregnancy at the beginning of the drug administration, kind of medication, period of exposure and dosage, deepest vertical pocket (DVP), and amniotic fluid index (AFI).
RESULTS
Of the 68 records identified, we analyzed 29 studies investigating the administration of NSAIDs, including 11 studies examined the administration of the Indomethacin, four articles have focused on the use of Nimesulide, and only two manuscripts considered the use of Diclofenac. We found a strict correlation between the development of oligohydramnios and the use of NSAIDs. The oligohydramnios is reversible, and the normal amount of amniotic fluid is restored after the interruption of the treatment.
CONCLUSIONS
The use of NSAIDs should be considered when maternal benefits outweigh the potential fetal risk, at the lowest effective dose for shortest duration. Beyond 48 h of NSAIDs treatment, we consider ultrasound monitoring of amniotic fluid, and we suggest stopping therapy if a decline AFI is present.
Topics: Pregnancy; Female; Humans; Oligohydramnios; Amniotic Fluid; Anti-Inflammatory Agents, Non-Steroidal; Pregnancy Trimester, Third; Ultrasonography; Pregnancy Outcome
PubMed: 38092425
DOI: 10.1080/14767058.2023.2253956 -
Chinese Medical Journal Oct 2022Complete wound regeneration preserves skin structure and physiological functions, including sensation and perception of stimuli, whereas incomplete wound regeneration... (Review)
Review
Complete wound regeneration preserves skin structure and physiological functions, including sensation and perception of stimuli, whereas incomplete wound regeneration results in fibrosis and scarring. Amniotic fluid stem cells (AFSCs) would be a kind of cell population with self-renewing and non-immunogenic ability that have a considerable role in wound generation. They are easy to harvest, culture, and store; moreover, they are non-tumorigenic and not subject to ethical restrictions. They can differentiate into different kinds of cells that replenish the skin, subcutaneous tissues, and accessory organs. Additionally, AFSCs independently produce paracrine effectors and secrete them in exosomes, thereby modulating local immune cell activity. They demonstrate anti-inflammatory and immunomodulatory properties, regulate the physicochemical microenvironment of the wound, and promote full wound regeneration. Thus, AFSCs are potential resources in stem cell therapy, especially in scar-free wound healing. This review describes the biological characteristics and clinical applications of AFSCs in treating wounds and provide new ideas for the treatment of wound healing.
Topics: Humans; Amniotic Fluid; Wound Healing; Regeneration; Skin; Cicatrix; Stem Cells
PubMed: 36535008
DOI: 10.1097/CM9.0000000000002076 -
The Journal of Maternal-fetal &... Jan 2022neurologic injury in myelomeningocele (MMC) occurs via a two-hit process: failed neural tube closure followed by neurodegeneration . Meconium in the amniotic fluid...
BACKGROUND
neurologic injury in myelomeningocele (MMC) occurs via a two-hit process: failed neural tube closure followed by neurodegeneration . Meconium in the amniotic fluid contains pancreatic digestive enzymes and is neurotoxic in rat models of MMC.
OBJECTIVES
The objectives of this study were to demonstrate the neurotoxicity of α-amylase and to compare the enzyme concentration and activity in the amniotic fluid of rats with retinoic acid induced MMC to a healthy control population.
STUDY DESIGN
Timed pregnant Sprague Dawley rats were gavage fed all-trans retinoic acid (60 mg/kg) in olive oil on gestational day E10 to induce a MMC defect. Control rats received olive oil. Amniotic fluid was collected on embryonic days E15, E17, E19, and E21. The amniotic fluid amylase concentration and relative activity were measured at each gestational age, and levels were compared between the MMC and control groups using Wilcoxon Rank Sum and Kruskal-Wallis tests. In a subset of dams sacrificed on E10.5, neuroepithelial cells were isolated from control embryos and exposed to α-amylase in increasing concentrations. Percentage of cell survival was assessed with CellProfiler software.
RESULTS
Amniotic fluid amylase activity for embryonic days E15, E17, E19, and E21 was determined for MMC and control pups. Amylase activity increased significantly from E15 to E21 in both control ( = 3.0 × 10) and MMC ( = 1.5 × 10) groups. Relative amylase activity was significantly increased in MMC pups compared to controls on E19 (247,792.8 versus 106,263.6; = .0019) and E21 (772,645.8 versus 481,975.3; = .021); no difference was detected on E15 (36,646.8 versus 40,179.3; = .645) or E17 (121,617.5 versus 71,750; = 1.000). , amylase demonstrated dose-dependent toxicity to fetal rat neuroepithelial cells.
CONCLUSION
Amylase concentration and activity level were higher in the amniotic fluid of rats with retinoic acid induced MMC compared to controls with advancing gestational age. As amylase is toxic to neural epithelial cells, the higher activity of this digestive enzyme in fetuses with MMC may be a contributor to neural tube damage . Future research should focus on amylase and other digestive enzymes in human MMC, as they may serve as potential targets of therapy.
Topics: Amniotic Fluid; Amylases; Animals; Female; Meningomyelocele; Pregnancy; Rats; Rats, Sprague-Dawley; Tretinoin
PubMed: 31910702
DOI: 10.1080/14767058.2020.1713082