Effects of simulated bleeding in an in vitro nasal fibroblast wound healing model.
BACKGROUND
We investigated the effect of simulated bleeding on plasminogen activity, matrix metalloproteinase (MMP) expression, and wound healing using a human fibroblast model.


METHODS
Nasal fibroblasts from three chronic rhinosinusitis (CRS) patients with nasal polyps and three controls were grown in culture and a standardized injury was created using a punch. To mimic bleeding, fibroblasts were stimulated with plasminogen (100 microg/mL), plasminogen + tranexamic acid (TA; 100 microg/mL) or media only. At 24, 48, and 72 hours after injury, we measured urokinase plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) activities and inactive and active MMP-2 and -9 expression.


RESULTS
Injury stimulated the nasal fibroblasts to express uPA and tPA and active and inactive MMP-2 and -9. In CRS patients, plasminogen significantly decreased MMP-9 expression after 48 hours (p < 0.04). In untreated fibroblasts, we observed a decrease in active MMP-9 expression, whereas plasminogen increased active MMP-9 expression after 48 hours (p < 0.04). At 24 hours, active MMP-9 expression was reduced by plasminogen +/- TA (p < 0.02). Plasminogen also stimulated uPA expression in CRS patient fibroblasts after 48 hours (p < 0.04). Fibroblast proliferation occurred when exposed to plasminogen and was strongly modulated by uPA and inactive and active MMP-2. The quality of wound healing was affected by inactive MMP-2, uPA and tPA, simulation, and inhibition of bleeding.


CONCLUSIONS
Activation of the plasminogen pathway and inactive MMP-2 expression tended to increase both proliferation of nasal fibroblasts and MMP-9 expression as a marker for deterioration of the quality of wound healing.
Department of Otorhinolaryngology, Head and Neck Surgery, University of Greifswald, Greifswald, Germany.
Achim Georg
AG
Beule
Theodore
T
Athanasiadis
John
J
Field
Werner
W
Hosemann
Peter-John
PJ
Wormald
Lor Wai
LW
Tan
D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002908:Chronic Disease; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006470:Hemorrhage / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D009666:Nose / Q000293:injuries* / Q000473:pathology; D010958:Plasminogen / Q000494:pharmacology; D012220:Rhinitis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D012852:Sinusitis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics; D014148:Tranexamic Acid / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics; D014945:Wound Healing / Q000187:drug effects
D015415:Biomarkers; D014148:Tranexamic Acid; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9