Intrauterine infection, such as occurs in chorioamnionitis, is a principal cause of preterm birth and is a strong risk factor for neurological morbidity and cerebral palsy. This study aims to examine whether human amnion epithelial cells (hAECs) can be used as a potential therapeutic agent to reduce brain injury induced by intra-amniotic administration of lipopolysaccharide (LPS) in preterm fetal sheep. Pregnant ewes underwent surgery at approximately 110 days of gestation (term is approx. 147 days) for implantation of catheters into the amniotic cavity, fetal trachea, carotid artery and jugular vein. LPS was administered at 117 days; hAECs were labeled with carboxyfluorescein succinimidyl ester and administered at 0, 6 and 12 h, relative to LPS administration, into the fetal jugular vein, trachea or both. Control fetuses received an equivalent volume of saline. Brains were collected 7 days later for histological assessment of brain injury. Microglia (Iba-1-positive cells) were present in the brain of all fetuses and were significantly increased in the cortex, subcortical and periventricular white matter in fetuses that received LPS, indicative of inflammation. Inflammation was reduced in fetuses that received hAECs. In LPS fetuses, the number of TUNEL-positive cells was significantly elevated in the cortex, periventricular white matter, subcortical white matter and hippocampus compared with controls, and reduced in fetuses that received hAECs in the cortex and periventricular white matter. Within the fetal brains studied there was a significant positive correlation between the number of Iba-1-immunoreactive cells and the number of TUNEL-positive cells (R2 = 0.19, p < 0.001). The administration of hAECs protects the developing brain when administered concurrently with the initiation of intrauterine inflammation.

Keywords:
Human amnion epithelial cells, Fetal brain injury, Intrauterine inflammation
1.
Goldenberg RL, et al: Epidemiology and cau-ses of preterm birth. Lancet 2008;371:75-84.
2.
Wu YW: Systematic review of chorioamnionitis and cerebral palsy. Ment Retard Dev Disabil Res Rev 2002;8:25-29.
3.
Wu YW, Colford JM Jr: Chorioamnionitis as a risk factor for cerebral palsy: a meta-analysis. JAMA 2000;284:1417-1424.
4.
Woodward LJ, et al: Neonatal MRI to predict neurodevelopmental outcomes in preterm infants. N Engl J Med 2006;355:685-694.
5.
Volpe JJ: Neurobiology of periventricular leukomalacia in the premature infant. Pediatr Res 2001;50:553-562.
6.
Leviton A, Paneth N: White matter damage in preterm newborns - An epidemiologic perspective. Early Hum Dev 1990;24:1-22.
7.
Dammann O, Leviton A: Maternal intrauterine infection, cytokines, and brain damage in the preterm newborn. Pediatr Res 1997;42:1-8.
8.
Yoon BH, et al: Amniotic fluid inflammatory cytokines (interleukin-6, interleukin-1beta, and tumor necrosis factor-alpha), neonatal brain white matter lesions, and cerebral palsy. Am J Obstet Gynecol 1997;177:19-26.
9.
Yoon BH, et al: High expression of tumor necrosis factor-alpha and interleukin-6 in periventricular leukomalacia. Am J Obstet Gynecol 1997;177:406-411.
10.
Peebles DM, et al: The effect of systemic administration of lipopolysaccharide on cerebral haemodynamics and oxygenation in the 0.65 gestation ovine fetus in utero. BJOG 2003;110:735-743.
11.
Mallard C, et al: White matter injury following systemic endotoxemia or asphyxia in the fetal sheep. Neurochem Res 2003;28:215-223.
12.
Duncan JR, et al: White matter injury after repeated endotoxin exposure in the preterm ovine fetus. Pediatr Res 2002;52:941-949.
13.
Gilles FH, Averill DR Jr, Kerr CS: Neonatal endotoxin encephalopathy. Ann Neurol 1977;2:49-56.
14.
Gilles FH, Leviton A, Kerr CS: Endotoxin leucoencephalopathy in the telencephalon of the newborn kitten. J Neurol Sci 1976;27:183-191.
15.
Nitsos I, et al: Chronic exposure to intra-amniotic lipopolysaccharide affects the ovine fetal brain. J Soc Gynecol Investig 2006;13:239-247.
16.
Kakishita K, et al: Human amniotic epithelial cells produce dopamine and survive after implantation into the striatum of a rat model of Parkinson's disease: a potential source of donor for transplantation therapy. Exp Neurol 2000;165:27-34.
17.
Okawa H, et al: Amniotic epithelial cells transform into neuron-like cells in the ischemic brain. Neuroreport 2001;12:4003-4007.
18.
Li H, et al: Immunosuppressive factors secreted by human amniotic epithelial cells. Invest Ophthalmol Vis Sci 2005;46:900-907.
19.
Ilancheran S, et al: Stem cells derived from human fetal membranes display multilineage differentiation potential. Biol Reprod 2007;77:577-588.
20.
Bailo M, et al: Engraftment potential of human amnion and chorion cells derived from term placenta. Transplantation 2004;78:1439-1448.
21.
Bose B: Burn wound dressing with human amniotic membrane. Ann R Coll Surg Engl 1979;61:444-447.
22.
Faulk WP, et al: Human amnion as an adjunct in wound healing. Lancet 1980;i:1156-1158.
23.
Parolini O, et al: Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells. Stem Cells 2008;26:300-311.
24.
Murphy S, et al: Human amnion epithelial cells prevent bleomycin-induced lung injury and preserve lung function. Cell Transplant 2011;20:909-923.
25.
Murphy S, et al: Amnion epithelial cell isolation and characterization for clinical use. Curr Protoc Stem Cell Biol 2010;1:1E6.
26.
Vosdoganes P, et al: Human amnion epithelial cells as a treatment for inflammation-induced fetal lung injury in sheep. Am J Obstet Gynecol 2011;205:156.e26-e33.
27.
Hodges RJ, et al: Human amnion epithelial cells reduce ventilation-induced preterm lung injury in fetal sheep. Am J Obstet Gynecol 2012;206:448.e8-e15.
28.
Back SA, Riddle A, Hohimer AR: Role of instrumented fetal sheep preparations in defining the pathogenesis of human periventricular white-matter injury. J Child Neurol 2006;21:582-589.
29.
Hansen AL, et al: Growth-plate chondrocyte cultures for reimplantation into growth-plate defects in sheep. Characterization of cultures. Clin Orthop Relat Res 1990;256:286-298.
30.
Duncan JR, et al: Chronic endotoxin exposure causes brain injury in the ovine fetus in the absence of hypoxemia. J Soc Gynecol Investig 2006;13:87-96.
31.
Dong W, et al: Treatment of intracerebral haemorrhage in rats with intraventricular transplantation of human amniotic epithelial cells. Cell Biol Int 2010;34:573-577.
32.
Jurewicz A, et al: Tumour necrosis factor-induced death of adult human oligodendrocytes is mediated by apoptosis inducing factor. Brain 2005;128:2675-2688.
33.
Sharief MK, Thompson EJ: In vivo relationship of tumor necrosis factor-alpha to blood-brain barrier damage in patients with active multiple sclerosis. J Neuroimmunol 1992;38:27-33.
34.
Wang X, et al: Lipopolysaccharide-induced inflammation and perinatal brain injury. Semin Fetal Neonatal Med 2006;11:343-353.
35.
Kramer BW, et al: Dose and time response after intraamniotic endotoxin in preterm lambs. Am J Respir Crit Care Med 2001;164:982-988.
36.
Czeh M, Gressens P, Kaindl AM: The yin and yang of microglia. Dev Neurosci 2011;33:199-209.
37.
Greisen G: Effect of cerebral blood flow and cerebrovascular autoregulation on the distribution, type and extent of cerebral injury. Brain Pathol 1992;2:223-228.
38.
Khwaja O, Volpe JJ: Pathogenesis of cerebral white matter injury of prematurity. Arch Dis Child Fetal Neonatal Ed 2008;93:F153-F161.
39.
Hutton LC, et al: Microglial activation, macrophage infiltration, and evidence of cell death in the fetal brain after uteroplacental administration of lipopolysaccharide in sheep in late gestation. Am J Obstet Gynecol 2008;198:117.e1-e11.
40.
Yang GY, et al: Tumor necrosis factor alpha expression produces increased blood-brain barrier permeability following temporary focal cerebral ischemia in mice. Brain Res Mol Brain Res 1999;69:135-143.
41.
Northington FJ, et al: Early neurodegeneration after hypoxia-ischemia in neonatal rat is necrosis while delayed neuronal death is apoptosis. Neurobiol Dis 2001;8:207-219.
42.
De Coppi P, et al: Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol 2007;25:100-106.
43.
Hao Y, et al: Identification of antiangiogenic and antiinflammatory proteins in human amniotic membrane. Cornea 2000;19:348-352.
© 2013 S. Karger AG, Basel
2013
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.