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Static Mechanical Loading Influences the Expression of Extracellular Matrix and Cell Adhesion Proteins in Vaginal Cells Derived From Premenopausal Women With Severe Pelvic Organ Prolapse

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Abstract

Introduction

Primary human vaginal cells derived from women with severe pelvic organ prolapse (POP-HVCs) demonstrate altered cellular characteristics as compared to cells derived from asymptomatic women (control-HVCs). Using computer-controllable Flexcell stretch unit, we examined whether POP-HVCs react differently to mechanical loading as compared to control-HVCs by the expression of extracellular matrix (ECM) components, cell–ECM adhesion proteins, and ECM degrading and maturating enzymes.

Methods

Vaginal tissue biopsies from premenopausal patients with Pelvic Organ Prolapse Quantification System stage ≥3 (n = 8) and asymptomatic controls (n = 7) were collected during vaginal hysterectomy or repair. Human vaginal cells were isolated by enzymatic digestion, seeded on collagen (COLI)-coated plates, and stretched (24 hours, 25% elongation). Total RNA was extracted, and 84 genes were screened using Human ECM and Adhesion Molecules polymerase chain reaction array; selected genes were verified by quantitative reverse transcription-polymerase chain reaction. Stretch-conditioned media (SCM) were collected and analyzed by protein array, immunoblotting, and zymography.

Results

In mechanically stretched control-HVCs, transcript levels of integrins (ITGA1, ITGA4, ITGAV, and ITGB1) and matrix metalloproteinases (MMPs) 2, 8, and 13 were downregulated (P < .05); in POP-HVCs, MMP1, MMP3, and MMP10, ADAMTS8 and 13, tissue inhibitor of metalloproteinases (TIMPs) 1 to 3, ITGA2, ITGA4, ITGA6, ITGB1, contactin (CNTN1), catenins (A1 and B1), and laminins (A3 and C1) were significantly upregulated, whereas COLs (1, 4, 5, 6, 11, and 12) and LOXL1 were downregulated. Human vaginal cells massively secrete MMPs and TIMPs proteins; MMP1, MMP8, MMP9 protein expression and MMP2 gelatinase activity were increased, whereas TIMP2 decreased in SCM from POP-HVCs compared to control-HVCs.

Conclusions

Primary human vaginal cells derived from women with severe pelvic organ prolapse and control-HVCs react differentially to in vitro mechanical stretch. Risk factors that induce stretch may alter ECM composition and cell–ECM interaction in pelvic floor tissue leading to the abatement of pelvic organ support and subsequent POP development.

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Authors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute at Mount Sinai Hospital, 25 Orde Street, Suite 6-1019, M5G 1X5, Toronto, Ontario, Canada

    Hala Kufaishi MD, MSc, Stephen Lye PhD & Oksana Shynlova PhD

  2. Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Mount Sinai Hospital, Toronto, Ontario, Canada

    May Alarab MD, MSc & Harold Drutz MD

  3. Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada

    May Alarab MD, MSc, Harold Drutz MD, Stephen Lye PhD & Oksana Shynlova PhD

Authors
  1. Hala Kufaishi MD, MSc
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  2. May Alarab MD, MSc
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  3. Harold Drutz MD
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  4. Stephen Lye PhD
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  5. Oksana Shynlova PhD
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Correspondence to Oksana Shynlova PhD.

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Kufaishi, H., Alarab, M., Drutz, H. et al. Static Mechanical Loading Influences the Expression of Extracellular Matrix and Cell Adhesion Proteins in Vaginal Cells Derived From Premenopausal Women With Severe Pelvic Organ Prolapse. Reprod. Sci. 23, 978–992 (2016). https://doi.org/10.1177/1933719115625844

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