(256) Cytokine expression in wound fluid: a pilot study

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Wound fluid cytokines may be one of many biological mechanisms that contribute to nociceptive sensitivity and high wound pain at rest and during wound care procedures. We plan to analyze the association of wound pain and the concentration of 10 wound fluid inflammatory and anti-inflammatory cytokines. Although wound fluid cytokines have been measured using a variety of techniques to collect and process the samples, little is known about the feasibility of using “direct” collection techniques rather than introducing absorptive wafers or other time-consuming techniques, such as aspirating fluid after covering the wound with semi-occlusive dressings. Direct methods are preferable to other techniques because the specimen does not need to be “treated” for the presence of the absorbent and/or the fluid is collected at the point of care. The amount of wound fluid that can be collected using direct methods in acute and chronic wounds is unknown. Further, it is not known whether small fluid samples can provide reliable results when measuring protein content and cytokine levels in a multi-plex context. The purpose of this pilot study is to 1) describe variation in the amount of fluid that can be collected using direct methods, 2) describe variation in the amount of protein content using the NanoDrop One Microvolume UV-Vis spectrophotometer, which only requires 1-2 μl of fluid to yield protein concentrations, and 3) describe the optimal dilution of fluid samples for multi-plex analyses. A three-fold serial dilution was performed to identify the ideal concentration for yielding accurate results for both acute and chronic wounds while maintaining sensitivity to detect samples on extreme ends of the spectrum. The findings of this study informed and helped to establish a standard for the collection and processing of wound fluid for cytokines and other mediators that may impact wound pain as well as wound healing.