Karch, JE, Baird, K, Fisher, T, Sommer, J, Zaluski, S, Brogan, M, Fries, K, Edsberg LE, Jaynes CD; Daemen College, Amherst, NY, U.S. mbrogan@daemen.edu.

PURPOSE: We have recently reported the development of a protocol for the biochemical analysis of proteins in wound fluids. This protocol incorporated gel electrophoresis, a technique that allows one to observe proteins as individual bands based on molecular weight. The purpose of this study was to identify these proteins. DESCRIPTION: Wound fluid samples were collected from patients (n>5, data continues to be collected) in a skilled nursing facility suffering with chronic wounds. The samples were taken with cotton swabs from the center of a wound and placed in phosphate-buffered saline. The proteins in these samples were analyzed by Western blot analysis for collagenase-1, transforming growth factor-β (TGF- β), platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF). We are currently investigating the presence of additional enzymes (collagenases) and growth factors (epidermal growth factor, fibroblast growth factor). It is expected that many of the bands generated by chronic wound fluid samples are growth factors and enzymes that, depending on their levels, promote or inhibit the healing process. Through identification and quantitation of protein bands generated from samples of healing and non-healing chronic wounds, we hope to establish the necessary relative levels of proteins that play a role in the healing process. OBSERVATIONS: Using Western blot analysis, we identified collagenase-1 and TGF- β in chronic wound fluid samples obtained from healing and non-healing wounds. Collagenase-1 concentration fluctuated initially, then decreased, in a healing wound but did not change in concentration in two non-healing wounds. TGF- β did not fluctuate in concentration at any time point in any wound. Although we did not detect the presence of PDGF or VEGF in our samples, we are looking for these growth factors in samples obtained from other patients. CONCLUSIONS: Through Western blot analysis, we have identified two proteins that may play a role in the healing process, collagenase-1 and TGF- β. Collagenase-1 is of particular interest because of the dramatic changes we observed in its concentration as the wound healed. Collagenase-1 remained at very high levels in two non-healing wounds throughout treatment. Fluctuation in the levels of this protein may indicate whether there is a biochemical response of the wound prior to any visual changes. FUNDING SOURCE: New York Chapter American Physical Therapy Association.

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