Neutrophil recruitment by fetal porcine endothelial cells: implications in scarless fetal wound healing.
Fetal dermal wounds heal scarlessly and with a minimal inflammatory response. When a robust inflammatory response is induced at the site of fetal dermal wounds by the application of cytokines, healing results in fibrosis. To test the hypothesis that the reduced inflammatory response in fetal wounds is due to impaired fetal leukocyte-endothelial interactions, the contributions of fetal endothelial cells to the inflammatory response in the fetus were investigated. Endothelial cells isolated from blood vessels of adult and mid-gestational fetal pigs were cultured in media until confluent monolayers were established. Adult porcine neutrophils were isolated and resuspended at a concentration of 1 million cells/mL. Interactions between neutrophils and endothelial cells were observed under static and flow conditions. Endothelial monolayers were exposed to neutrophils with and without prior stimulation of the endothelial cells with tumor necrosis factor alpha (TNF-alpha) for 4 h. The neutrophil-endothelial interactions were observed and analyzed for neutrophil adhesion, rolling velocity, and transmigration. Endothelial P-selectin mRNA expression was determined by real-time polymerase chain reaction (PCR). A novel in vitro model of fetal inflammation is described. Both adult and fetal endothelial cells demonstrated a dose-dependent increase in neutrophil adhesion and transmigration with increasing doses of TNF-alpha. The fetal response was significantly lower than the adult. As expected, rolling velocity was lower at higher cytokine concentrations and had an inverse correlation with P-selectin mRNA expression. Fetal endothelial cells are less permissive to adhesion and transmigration of neutrophils than adult endothelial cells. This may contribute to the paucity of inflammation seen in the fetal response to dermal injury.
Olutoye
Oluyinka O
OO
Michael E DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX 77030, USA. oolutoye@bcm.tmc.edu
Zhu
Xi
X
Cass
Darrell L
DL
Smith
C Wayne
CW
D000818:Animals; D002478:Cells, Cultured; D002921:Cicatrix / Q000276:immunology; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D005333:Fetus / Q000166:cytology*; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000276:immunology; D019007:P-Selectin / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D034421:Sus scrofa; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014945:Wound Healing / Q000276:immunology*
D019007:P-Selectin; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha