Inhibitory Effect of Progesterone on Cervical Tissue Formation in a Three-Dimensional Culture System with Human Cervical Fibroblasts1

House, Michael; Tadesse-Telila, Serkalem; Norwitz, Errol R.; Socrate, Simona; Kaplan, David L.

doi:10.1095/biolreprod.113.112540

Cited by 24 publications

(16 citation statements)

References 67 publications

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“…220 Herein the in vivo administration of vaginal progesterone increased MMP-9 activity and reduced collagen content in the cervical stroma. Vaginal progesterone also reduced the infiltration of active MMP-9-positive neutrophils and monocytes.…”

Section: Commentmentioning

confidence: 97%

Vaginal progesterone, but not 17α-hydroxyprogesterone caproate, has antiinflammatory effects at the murine maternal-fetal interface

Furcron

Romero

Plazyo

et al. 2015

American Journal of Obstetrics and Gynecology

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OBJECTIVE Progestogen (vaginal progesterone or 17-alpha-hydroxyprogesterone caproate [17OHP-C]) administration to patients at risk for preterm delivery is widely used for the prevention of preterm birth (PTB). The mechanisms by which these agents prevent PTB are poorly understood. Progestogens have immunomodulatory functions; therefore, we investigated the local effects of vaginal progesterone and 17OHP-C on adaptive and innate immune cells implicated in the process of parturition. STUDY DESIGN Pregnant C57BL/6J mice received vaginal progesterone (1 mg per 200 μL, n = 10) or Replens (control, 200 μL, n = 10) from 13 to 17 days postcoitum (dpc) or were subcutaneously injected with 17OHP-C (2 mg per 100 μL, n = 10) or castor oil (control, 100 μL, n = 10) on 13, 15, and 17 dpc. Decidual and myometrial leukocytes were isolated prior to term delivery (18.5 dpc) for immunophenotyping by flow cytometry. Cervical tissues were collected to determine matrix metalloproteinase (MMP)-9 activity by in situ zymography and visualization of collagen content by Masson’s trichrome staining. Plasma concentrations of progesterone, estradiol, and cytokines (interferon [IFN]-γ, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, KC/GRO, and tumor necrosis factor-α) were quantified by enzyme-linked immunosorbent assays. Pregnant mice pretreated with vaginal progesterone or Replens were injected with 10 μg of an endotoxin on 16.5 dpc (n = 10 each) and monitored via infrared camera until delivery to determine the effect of vaginal progesterone on the rate of PTB. RESULTS The following results were found: (1) vaginal progesterone, but not 17OHP-C, increased the proportion of decidual CD4+ T-regulatory cells; (2) vaginal progesterone, but not 17OHP-C, decreased the proportion of decidual CD8+CD25+Foxp3+ T cells and macrophages; (3) vaginal progesterone did not cause an M1→M2 macrophage polarization but reduced the proportion of myometrial IFNγ+ neutrophils and cervical active MMP-9-positive neutrophils and monocytes; (4) 17OHP-C did not reduce the proportion of myometrial IFNy-positive neutrophils; however, it increased the abundance of cervical active MMP-9-positive neutrophils and monocytes; (5) vaginal progesterone immune effects were associated with reduced systemic concentrations of IL-1β but not with alterations in progesterone or estradiol concentrations; and (6) vaginal progesterone pretreatment protected against endotoxin-induced PTB (effect size 50%, P = .008). CONCLUSION Vaginal progesterone, but not 17OHP-C, has local antiinflammatory effects at the maternal-fetal interface and the cervix and protects against endotoxin-induced PTB.

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Section: Commentmentioning

confidence: 97%

Vaginal progesterone, but not 17α-hydroxyprogesterone caproate, has antiinflammatory effects at the murine maternal-fetal interface

Furcron

Romero

Plazyo

et al. 2015

American Journal of Obstetrics and Gynecology

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“…One example of a method that offers the potential for significant advancement is the development of a hormonally regulated three-dimensional (3D) human cervical stromal cell culture system. 52 An advantage over the traditional cell culture system is the ability to assess the impact of the ECM on stromal function and vice versa. A 3D Fig.…”

Section: Development Of Tools and Techniques To Assess Cervical Changesmentioning

confidence: 99%

Distinct Roles of Cervical Epithelia and Stroma in Pregnancy and Parturition

Nallasamy

Mahendroo

2017

Semin Reprod Med

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Through pregnancy the cervix must simultaneously remain competent for pregnancy maintenance and yet become progressively compliant to ensure on time parturition. Cervical changes precede not only term but also preterm birth. Thus, an understanding of the molecular mechanisms by which the cervix maintains the delicate balance between competence and compliance is required to prevent the potential for lifelong health complications that can result from a premature birth. Recent advances and accumulating evidence support distinct roles for the cervical epithelia and stroma in sustaining competence. Concurrently, structural reorganization of the stromal extracellular matrix allows for the gradual decline in tissue compliance. In recent years, advances in our understanding of the cervical remodeling process has resulted from the collective insights derived from biological, genomics, engineering, and mathematical modeling studies on clinical samples and animal models. This review will highlight recent literature that advances understanding of (1) the importance of barrier function in the lower female reproductive tract in protection against ascending infection, (2) cellular and extracellular matrix changes in the cervical stroma that influence the mechanical function of the cervix, (3) the potential translation of biological insights into clinical tools that impact preterm birth, and (4) the distinction between term and specific pathways of preterm birth. Finally, we present a discussion of future areas of investigation that are likely to advance understanding and lead to the development of clinical tools for accurate detection and prevention of premature birth.

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“…Previously, a hormonally responsive, three-dimensional (3D) engineered cervix was developed using human cervical fibroblasts and porous silk protein scaffolds. [19][20][21] This engineered cervix produced 3D ECM that was inhibited after exposure to progesterone. 20 Inhibition of ECM formation was mitigated by the addition of mifepristone, a progesterone receptor antagonist, suggesting the involvement of the progesterone receptor.…”

Section: Introductionmentioning

confidence: 99%

“…[19][20][21] This engineered cervix produced 3D ECM that was inhibited after exposure to progesterone. 20 Inhibition of ECM formation was mitigated by the addition of mifepristone, a progesterone receptor antagonist, suggesting the involvement of the progesterone receptor. However, no data correlating ECM inhibition to changes in softness were presented.…”

Section: Introductionmentioning

confidence: 99%

Mechanical and Biochemical Effects of Progesterone on Engineered Cervical Tissue

House

Kelly

Klebanov

et al. 2018

Tissue Engineering Part A

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Preterm birth is a leading cause of morbidity and mortality in newborns. Babies born prematurely are at increased risk of lifelong health problems, including neurodevelopmental abnormalities. Cervical shortening precedes preterm birth in many women. Cervical shortening is caused, in part, by excessive softening of the extracellular matrix (ECM) of the cervical stroma. In clinical obstetrics, cervical shortening prompts treatment with supplemental progesterone to prevent preterm birth. However, progesterone-mediated effects on the cervical ECM are not well understood. This research sought to study progesterone-mediated remodeling of ECM produced by human cervical fibroblasts in vitro. A previously developed three-dimensional (3D) engineered model of the cervical ECM was used for experiments. Cervical fibroblasts were seeded on porous scaffolds and cultured in spinner flasks to promote ECM synthesis. Scaffolds were exposed to two conditions: 10 M estradiol versus 10 M estradiol +10 M progesterone for 4 weeks. To measure ECM strength, two scaffolds were mounted end-to-end on a wire and cultured such that ECM filled the gap between the scaffolds. The force required to pull the scaffolds apart was measured. Collagen content and collagen crosslinks were measured with ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry. Whole-transcriptome RNA sequencing (RNA-seq) was used to quantify gene expression between the two experimental conditions. Zymography was used to study the quantity and activity of matrix metalloproteinase-2 (MMP2) in the scaffolds. The study found that exposure to progesterone increased tissue softness of the engineered ECM over 28 days. Increased tissue softness correlated with decreased collagen content. With RNA-seq, progesterone exposure resulted in gene expression changes consistent with known progesterone effects. Pathway analysis of the RNA-seq data suggested MMPs were significantly dysregulated in progesterone-exposed engineered ECM. Increased expression of active MMP2 was confirmed in the progesterone-exposed engineered ECM. In summary, progesterone increased the softness of the ECM, which was correlated with decreased collagen production and altered histology. These results are important for deciphering the role of progesterone in preventing preterm birth.

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Inhibitory Effect of Progesterone on Cervical Tissue Formation in a Three-Dimensional Culture System with Human Cervical Fibroblasts1

Cited by 24 publications

References 67 publications

Vaginal progesterone, but not 17α-hydroxyprogesterone caproate, has antiinflammatory effects at the murine maternal-fetal interface

Vaginal progesterone, but not 17α-hydroxyprogesterone caproate, has antiinflammatory effects at the murine maternal-fetal interface

Distinct Roles of Cervical Epithelia and Stroma in Pregnancy and Parturition

Mechanical and Biochemical Effects of Progesterone on Engineered Cervical Tissue

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