In vivo localisation and vascular effects of nano vesicles derived from normal first trimester human placentae
s Placenta 45 (2016) 63e133 67 parameters. However in STs from OB women, 7,8 e DHF increased basal (103% p<0.02) ATP coupled (144% p<0.02) maximal (273% p<0.001) spare capacity (125% p<0.02, non mitochondrial respiration (220% p<0.02) and spare capacity (125% p<0.02) in a concentration dependent manner. Surprisingly, TRKB antagonism had no effect on respiratory parameters other than attenuating spare capacity and ATP coupled respiration (ANA 12, 72% p<0.03; Cyclotraxin B, 74% p<0.02) in STs from LN women. Conclusion: Our data show that TRKB agonist 7,8 DHF improves mitochondrial respiration in STs suggesting that the obese phenotype in the placenta can be rescued by activation of TRKB. NI2.05 IN VIVO LOCALISATION AND VASCULAR EFFECTS OF NANO VESICLES DERIVED FROM NORMAL FIRST TRIMESTER HUMAN PLACENTAE Mancy Tong Joanna Stanley Qi Chen Jo James Peter Stone Larry Chamley 1 The University of Auckland, Auckland, New Zealand; 2 Liggins Institute, Auckland, New Zealand Objectives: The maternal cardiovascular system undergoes dramatic adaptations during early pregnancy but how these changes are induced is unclear. The syncytiotrophoblast extrudes extracellular vesicles into the maternal circulation which may contribute to such adaptations. In this study we investigated: 1) the distribution/localisation of placental nanovesicles (pNVs) in vivo, and 2) whether pNVs can affect maternal vascular function. Methods: Following culture of normal placental explants (8 11 weeks) pNVs were isolated by differential centrifugation, labelled with CellTrace Far Red DDAO SE and administered intravenously to CD1 mice at gestational day 12.5 (1mg/mL) Nano vesicles isolated from culture media, without placental explants, were injected as controls. Animals were euthanised 30 minutes (n1/46) or 24 hours (n1/46) post injection. Solid organs were imaged using an IVIS fluorescence Imager. Arterial function was assessed by wire myography. Results: Placental nano vesicles were localised to the lungs and liver of pregnant mice after 30 minutes; and to the lungs, liver and kidneys after 24 hours. Mesenteric arteries from mice exposed to pNV for 30 minutes showed increased vasodilation in response to acetylcholine (Two way ANOVA, p<0.05) This effect was significantly prevented by the addition of nitric oxide (NO) synthase inhibitor, L NAME (p<0.05) Placental nanovesicles did not affect uterine artery function. Conclusion: It has long been known that placental macro vesicles (syncytial nuclear aggregates) are trapped in the maternal lungs but the in vivo distribution of pNVs remains unknown. This study showed that pNVs are localised to specific maternal organs, depending on time since exposure and pNVs can induce endotheliumand NOdependent vasodilation of resistance vessels. Here we provide novel, in vivo evidence that vesicles from first trimester placentae are biologically active and may play a role in mediating maternal cardiovascular adaptations to normal pregnancy, a process which may be deregulated in obstetric diseases. NI2.06 VASCULAR ENDOTHELIAL GROWTH FACTOR A ADMINISTRATION RESCUES FETOPLACENTAL ENDOTHELIAL CELL DEFECTS SEEN IN SEVERE FETAL GROWTH RESTRICTION Karen Mestan Hong Xin Emily Su University of Colorado, Aurora, CO, USA; Northwestern University, Chicago, IL, USA Objective: We have previously found that human fetoplacental endothelial cells (ECs) from growth restricted fetuses with abnormal umbilical artery Doppler velocimetry (FGRadv) show impaired angiogenesis. These ECs also demonstrate defects in migration and EC EC adherence. From a molecular perspective, FGRadv ECs have lower expression of aryl hydrocarbon receptor nuclear translocator (ARNT 1/4 HIF1b) This low ARNT expression contributes to decreased vascular endothelial growth factor A (VEGFA) expression in FGRadv ECs. Thus, our objectives were: (1) To determine whether VEGFA is lower in vivo in FGRadv pregnancies, and (2) To investigate whether administration of VEGFA would rescue the EC migration and adherence defects. Methods: From an ongoing birth cohort, 36 preterm infants from 23 36 weeks gestational age were studied. These infants were either growthrestricted with normal umbilical artery Dopplers (FGR) FGRadv, or were appropriately grown, gestational age matched controls. Cord blood plasma VEGFA was measured with sandwich immunoassays through the Northwestern University Comprehensive Metabolic Core. Human fetoplacental ECs were also isolated from 6 FGRadv and 6 control subjects. Cells were subjected to wound scratch assays, immunofluorescence, VEGFA treatment, or ARNT RNA interference. One way ANOVAwith post hoc testing or Kruskal Wallis analyses were utilized, and p<0.05 was considered significant. Results: Cord blood VEGFA concentrations were significantly decreased in FGRadv fetuses in comparison to controls and FGR fetuses with normal umbilical artery Doppler studies (p1/40.03) Mechanistically, control ECs subjected to ARNT knock down and FGRadv ECs demonstrated impaired EC migration that was rescued by VEGFA treatment (p1/40.001) Administration of VEGFA also improved VE cadherin expression in control ECs after ARNT knock down and in FGRadv ECs. Conclusions: VEGFA cord blood concentrations are significantly lower in pregnancies complicated by FGRadv. When administered to FGRadv ECs or control ECs that have been subjected to ARNT knock down, VEGFA rescues the migrational and EC EC adherence defects seen as a result of low ARNT. NI3.01 3D PRINTED, BIOENGINEERED PLACENTA MODEL INCORPORATING DECELLULARIZED PLACENTAL EXTRACELLULAR MATRIX AND PRIMARY TROPHOBLASTS Che Ying Kuo Kelly Rhodes Hannah Baker Navein Arumugasaamy Helim Aranda Espinoza Rohan Fernandes Melissa Fries John Fisher Peter Kim 1 Fischell Department of Bioengineeirng, University of Maryland College Park, College Park, MD, USA; 2 Sheikh Zayed Institute for Pediatric Surgical Innovation, Children's National Health System, Washington, DC, USA; 3 School of Medicine and Helath Sciences, The George Washington University, Washington, DC, USA; Department of Obstetrics, Washington Hospital Center, Washington, DC,