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Effects of simulated bleeding in an in vitro nasal fibroblast wound healing model

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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 μg/mL), plasminogen + tranexamic acid (TA; 100 μg/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.

Conclusion:

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.
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Keywords: Bleeding; chronic rhinosinusitis; endoscopic sinus surgery; fibroblasts; matrix metalloproteinases; plasminogen; tranexamic acid; wound healing model

Document Type: Research Article

Affiliations: Department of Otorhinolaryngology, Head and Neck Surgery, University of Greifswald, Greifswald, Germany; Department of Surgery–Otolaryngology Head and Neck Surgery, The Queen Elizabeth Hospital, Discipline of Surgery, The University of Adelaide, Australia

Publication date: 2010-05-01

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