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Clinical Research
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A simple method for acute wound fluid collection Gary Fudem, Ronald A. Ignotz, Timothy Roth, Heather Strom, Paul Savoie, Breda Cullen, Janice Lalikos, Raymond Dunn Numerous reports describe the presence of a variety of growth factors in wound fluid, yet no single work systematically examines multiple factors over time within an individual patient. We describe a simple, effective method of collecting wound fluid from skin graft donor sites for characterization of its components. Patients undergoing split-thickness skin grafts were enrolled in an IRB approved study to characterize wound fluid in a temporal study. After obtaining informed consent, split-thickness skin grafts were harvested with standard clinical procedures. Immediately after harvesting the skin graft, the donor site was covered with a non-adherent gauze bandage* for approximately 1 hour. Following removal of the bandage, the donor site was covered with a transparent dressing**. Fluid collected under the dressing to create a ÒblisterÓ maintaining a moist wound-healing environment. Periodically, wound fluid was drawn off with a syringe. This process could be repeated over a five-day period with most patients before fluid production had subsided to levels insufficient to collect. Once collected, fluid was clarified by centrifugation at 10,000-x g, aliquoted and stored at -80oC. Quantization of fluid collected per square centimeter of donor site shows that fluid production peaked at 48 hours with sufficient fluid collected for at least five days. We have analyzed fluid for the temporal appearance of several growth factors. ELISA data shows that PDGF-AA was detected at the earliest points then rapidly declined. KGF concentration was low at the earliest times, rapidly increased for three days then began to decline. Data represents results from 7 patients normalized to albumin content. Other assays can be performed depending on the investigator's interests. Lastly, a welcome benefit to the transparent dressing is that all the patients reported no pain in the postoperative period related to the donor site .*Release Dressing, **Bioclusive Dressing (Johnson and Johnson) |
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