SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 172A

172A

Reproductive Sciences Vol. 25, Supplement 1, March 2018

hypothesis. This reduction in endothelial GC barrier function was
accompanied by a higher level of a circulating component of the
glycocalyx (Sdc1) prior to delivery in NL. The physiologic significance
of these pregnancy differences in the GC remain to be determined.
*Figure(s) will be available online.

T-188
High D-Glucose and TNF-α-Induced Endothelial Cell Dysfunction
and Activation in Human Umbilical Vein Endothelial Cells. Tamara
Sáez†,1,2 Paul de Vos*,2 Bart de Haan,2 Luis Sobrevia*,1 Marijke M Faas*.2
1
Pontificia Universidad Católica de Chile, Santiago, Chile; 2University
of Groningen, Groningen, Netherlands.
INTRODUCTION: Gestational diabetes mellitus (GDM) is characterized
by fetoplacental endothelial cell dysfunction (increased endothelial nitric
oxide synthase (eNOS) and human cationic amino acid type 1 (hCAT-1)
expression), and activation (increased intracellular adhesion molecule
type 1 (ICAM-1) expression). This may be due to high levels of glucose
and a proinflammatory state in fetoplacental circulation. We hypothesized
that high D-glucose (HG) and the proinflammatory tumor necrosis alpha
(TNF-α) induce endothelial dysfunction and activation in human umbilical
vein endothelial cells (HUVECs) characteristic for GDM.
METHODS: HUVECs were exposed to HG (25 mmol/L) and/or TNF-α
(2 ng/mL) during 24 h and endothelial cell function and activation were
evaluated by wound healing assay, expression of eNOS, hCAT-1, vascular
endothelial growth factor (VEGF) (qPCR) and ICAM-1 (flow cytometry)
and cytokine release (IL6, IL8, IL10 and MCP-1) (affymetrix assay).
RESULTS: HG increased endothelial cell migration, increased eNOS
and hCAT-1 mRNA expression (p<0.05), but did not affect VEGF mRNA
expression, slightly increased percentage ICAM-1 positive cells and
only increased the release of IL8. TNF-α did not affect endothelial cell
migration, eNOS, hCAT-1 and VEGF expression; it highly increased the
percentage of ICAM-1 positive cells and the release of IL6, IL10 and
MCP-1 (p<0.05). The coincubation of HUVECs with TNF-α and HG
decreased endothelial cell migration, decreased eNOS mRNA expression,
did not affect hCAT-1 mRNA (p>0.05) expression and increased VEGF
mRNA expression (p<0.05) compared with HUVEC-HG. It also highly
increased the percentage of ICAM-1 expressing cells and the release of
IL6, IL8, IL10 and MCP-1 (p<0.05) compared with HUVEC-HG.
CONCLUSION: TNF-α and HG together may be involved in inducing
fetoplacental endothelial dysfunction and activation in GDM. However,
they do not act synergistically but TNF-α may induce endothelial cell
activation, while HG may induce endothelial dysfunction.

T-189
In Vitro Chronic Exposure to Ureplasma urealyticum Modifies Innate
Immune Expression Genes in Human Fetal Membranes.. Damaris
Ortega-Pérez†,1,3 Silvia Giono-Cerezo*,1 Maribel Sánchez-Martínez*,3
Fabian Arechavaleta-Velasco*,2 Aurora Espejel-Nuñez*,3 Guadalupe
Estrada-Gutiérrez*.3 1Escuela Nacional de Ciencias Biológicas, Instituto
Politécnico Nacional, Mexico City, Mexico; 2IMSS, Mexico City, Mexico;
3
Instituto Nacional de Perinatología, Mexico City, Mexico.
INTRODUCTION: Intrauterine infection is the main risk factor for
premature rupture of membranes. This pathology complicates 10%
of pregnancies around the world. During an ascending infection, the
choriodecidua is the first-line barrier in contact with pathogens, which
cross fetal membranes leading to chorioamnionitis. Ureaplasma is
a common isolated microorganism and it is strongly associated with
premature rupture of membranes and prematurity. In animal models, it
has been shown that Ureaplasma elicits a robust innate immune response
leading to preterm labor. However, the molecular processes underlying
chronic infection by this microorganism in human fetal membranes
remains unknown. The aim of the study was to determine if in vitro chronic
infection by Ureaplasma urealyticum (Uu) modifies innate immune
expression genes in human fetal membranes.
METHODS: Fetal membranes (n=7) were collected from pregnancies at
term (37-39 wk) delivered by c-section without evidence of active labor or
infection. The tissues were washed with PBS 1X to remove adherent blood
clots, and mounted in a transwell device with the choriodecidua facing

Scientific Abstracts

the upper chamber and the amnion facing the lower chamber, making
it possible to test the two compartments independently. Choriodecidual
tissues were stimulated with 1x106 Uu color changing units during 120h,
simulating an ascending infection. Culture media and bacteria were
changed every 24h to maintain cellular and bacterial viability. mRNA was
extracted from amnion and choriodecidua separately, testing quantity and
quality using an Agilent Bioanalyzer, and Human Antibacterial Response
qRT-PCR Arrays (SABioscience-Qiagen) were perfomed, including 84
key genes related to innate immunity.
RESULTS: Choriodecidual tissues exposed to chronic Uu infection shows
13 genes with modified expression (9 upregulated and 4 downregulated).
In infected amnion 15 genes change expression comparing to noninfected controls (9 upregulated and 6 downregulated). For example:
TLR9 is upregulated choriodecidua infection, in contrast this gene is
downregulated in amnion; IL12B and MEFV are upregulated both,
choriodecidua and amnion.
CONCLUSION: Our results show, for the first time, that Uu modifies
the innate immune response in human fetal membranes during chronic
infection; this response is different in each tissue of the fetal membranes.
Ongoing analysis focused on each gene of interest could reveal their
role during an Uu infection and consequently, in premature rupture of
membranes.

T-190
Serum Insulin-Like Growth Factor Binding Protein Levels Predict
IVF Outcome. Fabiana M Kreines†, Mohamad Irani†, Dimitrios
Nasioudis†, Evelyn Minis, Steven S Witkin, Steven D Spandorfer*. Weill
Cornell Medicine, New York, NY, United States.
INTRODUCTION: Previous retrospective studies have suggested a
relationship between serum and follicular fluid levels of insulin-like
growth factor (IGF) and IGF-binding protein (IGFBP) and embryo
development and implantation. Specifically, it has been suggested that
higher levels of IGFBP in both serum and follicular fluid at various time
points are associated with oocyte maturation, better embryo development,
and positive IVF cycle outcome. We conducted this prospective study to
overcome the limitations of retrospective analyses and to better evaluate
a potential role for IGFBP in predicting IVF outcomes.
METHODS: Sera from 205 women undergoing an IVF cycle were
prospectively obtained on cycle days 24 and 26 (10 and 12 days after
oocyte retrieval, respectively) and assayed by ELISA for concentrations
of IGFBP. Cycle outcomes were prospectively followed. Associations
between IGFBP levels and outcome were evaluated by the Mann-Whitney
test.
RESULTS: The mean age of all subjects was 37.8±4.4 years, mean BMI
was 24.3±4.2 kg/m2, and mean harvested oocytes was 9.9±7.2. Twentyseven women (13.2%) had their cycles cancelled, 9 (4.4%) did not have
an embryo transfer, while 169 (82.4%) proceeded to embryo transfer.
Among patients who underwent an embryo transfer, 83 (49.1%) had an
initial positive pregnancy test, 71 (85.5%) of whom achieved a clinical
pregnancy and 52 (30.8%) reached an ongoing pregnancy (>24 weeks'
gestation). Patients with an initial positive pregnancy test had a higher
mean level of IGFBP on cycle day 26 compared with those who had a
negative test (20.9 vs. 14.5 ng/mL, P=0.04). Patients who subsequently
achieved a clinical pregnancy also had a higher IGFBP level on cycle
day 26 than did those without a clinical pregnancy (21.7 vs. 14.3 ng/mL,
P=0.02). Furthermore, patients who achieved an ongoing pregnancy
had a higher IGFBP level on cycle day 26 compared to those who had
negative outcomes (no pregnancy or pregnancy loss) (23.9 vs. 14.7 ng/
mL, P<0.01). There was no significant difference in serum IGFBP levels
measured on cycle day 24 between the groups.
CONCLUSION: Elevated serum IGFBP levels on day 26 of an IVF
cycle, 2 days before the first pregnancy test, is associated with subsequent
clinical and ongoing pregnancies. Additional studies on larger populations
are needed to verify this observation and establish a precise cutoff value
for serum IGBP that accurately predicts IVF outcome.

Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018