Inflammatory response of human gestational membranes to Ureaplasma parvum using a novel dual-chamber tissue explant system

Lauren C. Potts, Liping Feng, Patrick C. Seed, Friederike L. Jayes, Maragatha Kuchibhatla, Brian Antczak, Matthew K. Nazzal, Amy P. Murtha*

*Corresponding author for this work

Research output: Contribution to journalArticle

4 Citations (Scopus)

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. Dosedependent 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.

Original languageEnglish (US)
Article number119
JournalBiology of Reproduction
Volume94
Issue number5
DOIs
StatePublished - May 1 2016

Fingerprint

Ureaplasma
Amnion
Interleukin-8
Membranes
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Infectious Pregnancy Complications
Tissue Survival
Peptide Hydrolases
Coloring Agents
Enzyme-Linked Immunosorbent Assay
Cell Proliferation
Cytokines
Inflammation
Equipment and Supplies
Polymerase Chain Reaction
Infection
Preterm Premature Rupture of the Membranes

Keywords

  • Cytokines
  • Gestational membranes
  • Preterm premature rupture of membranes
  • Tissue explant
  • Ureaplasma parvum

ASJC Scopus subject areas

  • Cell Biology

Cite this

Potts, Lauren C. ; Feng, Liping ; Seed, Patrick C. ; Jayes, Friederike L. ; Kuchibhatla, Maragatha ; Antczak, Brian ; Nazzal, Matthew K. ; Murtha, Amy P. / Inflammatory response of human gestational membranes to Ureaplasma parvum using a novel dual-chamber tissue explant system. In: Biology of Reproduction. 2016 ; Vol. 94, No. 5.
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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. Dosedependent 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.",
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Inflammatory response of human gestational membranes to Ureaplasma parvum using a novel dual-chamber tissue explant system. / Potts, Lauren C.; Feng, Liping; Seed, Patrick C.; Jayes, Friederike L.; Kuchibhatla, Maragatha; Antczak, Brian; Nazzal, Matthew K.; Murtha, Amy P.

In: Biology of Reproduction, Vol. 94, No. 5, 119, 01.05.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Inflammatory response of human gestational membranes to Ureaplasma parvum using a novel dual-chamber tissue explant system

AU - Potts, Lauren C.

AU - Feng, Liping

AU - Seed, Patrick C.

AU - Jayes, Friederike L.

AU - Kuchibhatla, Maragatha

AU - Antczak, Brian

AU - Nazzal, Matthew K.

AU - Murtha, Amy P.

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AB - 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. Dosedependent 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.

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KW - Tissue explant

KW - Ureaplasma parvum

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