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

Reproductive Sciences
Hala KufaishiOksana Shynlova

Abstract

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. 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. In mechanically stretched control-HVCs, transcript levels of integrins (ITGA1, ITGA4, IT...Continue Reading

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Citations

Jul 13, 2017·Nature Reviews. Gastroenterology & Hepatology·George S HusseyStephen F Badylak
Jul 9, 2020·International Journal of Molecular Sciences·Alejandra M Ruiz-ZapataEgbert Oosterwijk
Aug 6, 2021·International Urogynecology Journal·Luiz Gustavo Oliveira BritoMaria Augusta T Bortolini

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Methods Mentioned

BETA
biopsies
hysterectomy
biopsy
dissection
SMA
protein assay
Profiler
PCR
antibody array
electrophoresis

Software Mentioned

Prism
Graph Pad
PCR array
Quantibody Q - Analyzer
ImageJ
CFX Manager
ArrayVision
geNorm
GraphPad
Flexcell

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