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Molecular characterization of fibroblasts isolated from human peritoneum and adhesions
G M Saed1, W Zhang, M P Diamond
1Department of Obstetrics and Gynecology, Hutzel Hospital, Wayne State University, Detroit, Michigan, USA. g.saed@wayne.edu
Objective:
To determine the response of adhesion and peritoneal fibroblasts to hypoxia.
Design:
Prospective experimental study.
Setting:
University medical center.
Patient(S):
Primary cultures of fibroblasts established from the peritoneal and adhesion tissues of the same patients (n = 2) to minimize genetic variations.
Intervention(S):
Hypoxia treatment of the primary cultured fibroblast.
Main Outcome Measure(S):
Analyze the expression of extracellular matrix (ECM) components, metalloproteinases and their tissue inhibitors, growth factors, and cytokines in adhesion and peritoneal fibroblasts under normal and hypoxic conditions by reverse transcriptase/polymerase chain reaction analysis.
Result(S):
Compared to peritoneal fibroblasts, adhesion fibroblasts had a significant increase in the basal mRNA levels for collagen I, fibronectin, MMP-1, TIMP-1, TGF-beta 1, TGF-beta 2, and IL-10. Hypoxia resulted in a further increase in collagen 1, fibronectin, TIMP-1, TGF-beta 1, TGF-beta 2, IL-10, and IFN-gamma mRNA levels in both peritoneal and adhesion fibroblasts. The increase was more profound in adhesion fibroblasts.
Conclusion(S):
Hypoxia induces molecular changes in both peritoneal and adhesion fibroblasts, creating a milieu that favors adhesion development. The effect of hypoxia was more profound on adhesion fibroblasts.