title: Inflammatory Response of Human Gestational Membranes to Ureaplasma parvum Using a Novel Dual-Chamber Tissue Explant System.
abstract: Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy.
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina.
authors.forename: Lauren C
authors.initials: LC
authors.lastname: Potts
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina.
authors.forename: Liping
authors.initials: L
authors.lastname: Feng
authors.affiliation: Department of Pediatrics, Duke University, Durham, North Carolina.
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Seed
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina Department of Pathology, Duke University, Durham, North Carolina.
authors.forename: Friederike L
authors.initials: FL
authors.lastname: Jayes
authors.affiliation: Department of Biostatistics and Bioinformatics, Duke University, Durham, North Carolina.
authors.forename: Maragatha
authors.initials: M
authors.lastname: Kuchibhatla
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina.
authors.forename: Brian
authors.initials: B
authors.lastname: Antczak
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina.
authors.forename: Matthew K
authors.initials: MK
authors.lastname: Nazzal
authors.affiliation: Department of Obstetrics and Gynecology, Duke University, Durham, North Carolina Department of Pediatrics, Duke University, Durham, North Carolina murth002@mc.duke.edu.
authors.forename: Amy P
authors.initials: AP
authors.lastname: Murtha
mesh_terms: D000650:Amnion / Q000378:metabolism; D002821:Chorioamnionitis / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology*; D016207:Cytokines / Q000378:metabolism; D005321:Extraembryonic Membranes / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008954:Models, Biological; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000378:metabolism / Q000382:microbiology / Q000473:pathology*; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods*; D014509:Ureaplasma / Q000302:isolation & purification / Q000502:physiology*; D016869:Ureaplasma Infections / Q000378:metabolism / Q000473:pathology*
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators
keywords: Ureaplasma parvum; cytokines; gestational membranes; preterm premature rupture of membranes; tissue explant