Effects of oxidized regenerated cellulose on the expression of extracellular matrix and transforming growth factor-beta1 in human peritoneal fibroblasts and mesothelial cells.
OBJECTIVE
The purpose of this study was to evaluate the potential biologic effects of oxidized regenerated cellulose, which has been shown in multiple human in vivo studies to reduce postoperative adhesion development, on the messenger RNA levels of transforming growth factor-beta1, type I collagen, type III collagen, and fibronectin.


METHODS
The oxidized regenerated cellulose was dissolved in saline solution and added to confluent, monolayer cultures of human normal fibroblasts and mesothelial cells. Control cells were maintained in media alone at the same pH. After 24 hours of treatment, total RNA was extracted from all cells. Real-time reverse transcription-polymerase chain reaction was performed to determine the relative change in messenger RNA levels of type I, type HI collagen, fibronectin, transforming growth factor-beta1, and beta-actin (housekeeping gene) in response to the oxidized regenerated cellulose treatment (n=4 cultures). Student t tests were performed for each cell type, which compared oxidized regenerated cellulose-treated cells to control cells. Calculated power for the statistically significant findings ranged from 65% to 100%.


RESULTS
Transforming growth factor-beta1 messenger RNA was elevated by the oxidized regenerated cellulose treatment in the mesothelial cells by 13% (control cells, 0.562+/-0.022; oxidized regenerated cellulose-treated cells, 0.636+/-0.014; P=.03). In normal fibroblasts, transforming growth factor-beta1 messenger RNA was slightly, but not significantly, decreased in oxidized regenerated cellulose-exposed normal fibroblasts compared with controls (control cells, 0.622+/-0.062; oxidized regenerated cellulose-treated cells, 0.609+/-0.006; P=.85). Type I collagen was found to be increased by exposure to oxidized regenerated cellulose in both mesothelial cells and normal peritoneal fibroblasts. Type I collagen was increased by 23% in mesothelial cells (control cells [0.587+/-0.018] vs oxidized regenerated cellulose-treated cells [0.722+/-0.010], P=.002), and 27% in normal fibroblasts (control cells, 0.522+/-0.018, oxidized regenerated cellulose-treated cells, 0.665+/-0.009; P=.001). However, messenger RNA levels of type III collagen and fibronectin (other representative molecules of the extracellular matrix) were not altered significantly by oxidized regenerated cellulose application in vitro.


CONCLUSIONS
Oxidized regenerated cellulose influences the expression of factors that are accepted commonly to be associated with adhesiogenesis. Oxidized regenerated cellulose increased the expression of transforming growth factor-beta1 in mesothelial cells and type I collagen in mesothelial cells and normal peritoneal fibroblasts. Minimization of these oxidized regenerated cellulose-induced molecular changes, if they occur in vivo, may improve the ability of oxidized regenerated cellulose to reduce postoperative adhesion development.
Gago
L April
LA
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Wayne State University School of Medicine, 4707 St. Antoine Boulevard, Detroit, MI 48201, USA. lagago@med.wayne.edu
Saed
Ghassan M
GM
Wang
Rona X
RX
Kruger
Michael
M
Diamond
Michael P
MP
D002478:Cells, Cultured; D002483:Cellulose, Oxidized / Q000494:pharmacology*; D018076:DNA, Complementary / Q000032:analysis; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D006801:Humans; D010537:Peritoneum / Q000166:cytology; D011336:Probability; D012333:RNA, Messenger / Q000032:analysis; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016212:Transforming Growth Factor beta / Q000187:drug effects / Q000378:metabolism*; D053773:Transforming Growth Factor beta1
D002483:Cellulose, Oxidized; D018076:DNA, Complementary; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1