0042. Burn injury stabilises extracellular atp and induces microvesicle production in skin
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ORAL PRESENTATION
Open Access
0042. Burn injury stabilises extracellular atp and induces microvesicle production in skin U Katbeh*, M Takata, KP O’Dea From ESICM LIVES 2014 Barcelona, Spain. 27 September - 1 October 2014 Introduction Microvesicles (MVs) are subcellular membrane-enclosed particles released from activated or dying cells with multiple functions, including induction of inflammation. A role for MVs has been indicated in sepsis- and traumarelated systemic inflammatory response syndrome (SIRS), but their production and function in severe burns injury is unknown. Extracellular ATP released from mechanically injured cells is an important stimulus for MV production [1], but this response is usually limited spatially as well as temporally by the rapid breakdown of extracellular ATP by ectoATPases. Due to the known sensitivity of ectoATPases to thermal inactivation [2], we hypothesised that ATP released during thermal injury would be stable and capable of inducing MVs from viable responder cells within the burn tissue microenvironment. Objectives To investigate the release of ATP from thermallyinjured cells and its potential role in inducing MV release from burn-injured tissue.
Results ATP was released by thermal injury to keratinocytes and remained stable for up to 3 hours at 37°C (Fig. 1). In contrast, ATP released by freeze-thawing decayed rapidly. Incubation of skin explants with ATP produced
Figure 1 n=4-7
Methods In vitro cultured human keratinocytes were injured thermally by incubation at 95°C for up to 60 seconds, or mechanically by one freeze-thaw cycle. ATP release was quantified using a fluorimetric assay. An ex vivo mouse skin explant model was developed in which skin pieces were exposed to steam for 15 or 60 secs, or to ATP (6mM) sub-dermally, and then incubated in medium for 2 hours at 37°C. Released MVs were analysed by flow cytometry. Skin oedema was assessed by weight gain during the incubation period.
Imperial College London, Section of Anaesthetics, Pain Medicine and Intensive Care, London, UK
Figure 2 n=4-7, *p
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