title: 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.
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, 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.
  
  
  
  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.
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: Hala
authors.initials: H
authors.lastname: Kufaishi
authors.affiliation: Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Mount Sinai Hospital, Toronto, Ontario, Canada Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada.
authors.forename: May
authors.initials: M
authors.lastname: Alarab
authors.affiliation: Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Mount Sinai Hospital, Toronto, Ontario, Canada Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Harold
authors.initials: H
authors.lastname: Drutz
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Stephen
authors.initials: S
authors.lastname: Lye
authors.affiliation: Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada Department of Obstetrics and Gynecology, University of Toronto, Toronto, Ontario, Canada shynlova@lunenfeld.ca.
authors.forename: Oksana
authors.initials: O
authors.lastname: Shynlova
mesh_terms: D000328:Adult; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D006801:Humans; D008875:Middle Aged; D056887:Pelvic Organ Prolapse / Q000378:metabolism*; D017697:Premenopause / Q000378:metabolism; D013314:Stress, Mechanical; D014621:Vagina / Q000378:metabolism* / Q000473:pathology / Q000502:physiology
chemical_list: D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins
keywords: POP; human vagina; mechanical stretch; premenopausal