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Laboratory Research
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Investigation into the role of proteases in extracellular matrix regulation A. Essler, BSC, C. Boyle, MSc, B. Cullen, PhD Wound repair is a carefully balanced dynamic process, involving both stimulatory and degradative factors in which any alteration may lead to delayed healing. The chronic wound environment has been demonstrated to contain elevated levels of degradative factors, specifically proteases, resulting in an imbalance in the healing process. Previous studies have indicated that the chronic wound environment contains elevated levels of matrix metalloproteinases (MMPs) and consequently they have been implicated in chronic wound pathophysiology..In this study we investigated the effect of elevated levels of MMPs on extracellular matrix (ECM) degradation, and the ability of collagen/ORC to negate this effect. Levels of MMP1, 2, 3, 7, 8 and 9 were quantified in acute and chronic wounds as these enzymes collectively have the ability to degrade major ECM components. The efficiency of ORC/collagen to bind and inactivate these proteases was also quantified. Techniques including substrate activity assays, zymography and ELISA were utilized to determine MMP levels. .Results demonstrated that MMP activity, specifically MMP-2 and 9 is significantly elevated in chronic tissue when compared to exudates. Analysis indicated that these elevated gelatinases were able to degrade ECM components, however this effect was corrected after pretreatment of the exudates with collagen/ORC. This suggests that collagen/ORC is able to protect endogenous extracellular proteins by redressing the proteolitic imbalance within the wound environment. |
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