• Wound Repair Regen · Sep 2004

    Comparative Study

    Expression of transforming growth factor-beta and extracellular matrix by human peritoneal mesothelial cells and by fibroblasts from normal peritoneum and adhesions: effect of Tisseel.

    • Ghassan M Saed, Michael Kruger, and Michael P Diamond.
    • Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Wayne State University/Detroit Medical Center, Hutzel Hospital, 4707 St. Antoine Boulevard, Detroit, MI 48201, USA.
    • Wound Repair Regen. 2004 Sep 1; 12 (5): 557-64.

    AbstractWe have previously shown that fibroblasts obtained from adhesions produce greater amounts of transforming growth factor-beta 1 (TGF-beta1) and extracellular matrix (ECM) molecules than normal fibroblasts isolated from normal peritoneum. The purpose of the current studies was to examine the effect of Tisseel (Baxter Healthcare Corporation, Glendale, CA), a fibrin sealant containing fibrinogen, aprotinin (a protease inhibitor), thrombin, and CaC1(2), on TGF-beta1 and ECM production by human peritoneal mesothelial cells, normal peritoneal fibroblasts, and adhesion fibroblasts. Multiplex reverse transcription-polymerase chain reaction using beta-actin as a housekeeping gene was used to determine mRNA levels of TGF-beta1 and ECM in these cells at 6, 12, 24, and 48 hours under normoxic conditions in the following treatment groups : fibrin sealant (Tisseel) alone; fibrin sealant with the two components diluted 1 : 2; fibrin sealant with the sealer protein component reconstituted without aprotinin (a protease inhibitor); fibrin sealant with the sealer protein component reconstituted without aprotinin (and both components diluted 1 : 2); fibrin sealant components diluted to physiologic concentrations; and control (culture media). The test compositions had little effect on TGF-beta1 mRNA expression in mesothelial cells and normal peritoneal fibroblasts, but resulted in a marked reduction of TGF-beta1 from adhesion fibroblasts. Expression of type I collagen by human peritoneal mesothelial cells was not detected; the compositions reduced type I collagen mRNA expression by both types of fibroblasts. Type III collagen was detected at six hours, and increased approximately 50 percent by culturing for 48 hours. Tisseel at full strength and with both components diluted 1 : 2 initially increased type III collagen mRNA levels; in contrast, type III collagen mRNA levels were reduced in mesothelial cells by the fibrin sealant without aprotinin at both concentrations and at physiologic concentrations. In both types of fibroblasts, the Tisseel compositions reduced type III collagen mRNA expression. Fibronectin mRNA were transiently reduced at six hours by approximately 50 percent in the presence of the Tisseel components, but then returned to control levels. Fibronectin mRNA levels were not altered in normal peritoneal fibroblasts, but were reduced by all but the physiologic concentration in adhesion fibroblasts. Tisseel may modulate human peritoneal mesothelial cell, normal peritoneal fibroblast, and adhesion fibroblast function. These results suggest that fibrin sealant prepared from the Tisseel kit without aprotinin has the ability to reduce ECM and TGF-beta1 mRNA levels, especially from adhesion fibroblasts, which may indicate a role in reduction of postoperative adhesion development.

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