Characterization of the Blister Fluid Proteome for Pediatric Burn Classification.

Blister fluid (BF) is a novel and viable research matrix for burn injury study, which can reflect both systemic and local microenvironmental responses. The protein abundance in BF from different burn severities were initially observed using a 2D SDS-PAGE approach. Subsequently, a quantitative data independent acquisition (DIA) method, SWATH, was employed to characterize the proteome of pediatric burn blister fluid. More than 600 proteins were quantitatively profiled in 87 BF samples from different pediatric burn patients. These data were correlated with clinically assessed burn depth and time until complete wound re-epithelialization through several different statistical analyses. Several proteins from these analyses exhibited significant abundance change between different burn depth or re-epithelialization groups, and can be considered as potential biomarker candidates. Further gene ontology (GO) enrichment analysis of the significant proteins revealed the most significant burn related biological processes (BP) that are altered with burn depth, including homeostasis and oxygen transport. However, for wounds with re-epithelialization times more or less than 21 days, the significant GO annotations were related to enzyme activity. This quantitative proteomics investigation of burn BF may enable objective classification of burn wound severity and assist with clinical decision-making. Data are available via ProteomeXchange with identifier PXD011102.

The blister fluid proteome of paediatric burns.

UNLABELLED: Burn injury is highly traumatic for paediatric patients, with the severity of the burn often dictating the extent of scar formation. The diagnosis of burn wound severity is largely determined by the attending clinician's experience. Thus, a greater understanding of the biochemistry at burn wound site environment and the biology of burns of different severities at an earlier stage may reduce the reliance on subjective diagnoses. In this study, blister fluid was collected from superficial thickness, deep-partial thickness, and full-thickness paediatric burn wounds. Samples were combined together based on burn depth classification and then subjected to four different fractionation methods followed by trypsin digestion. Peptides were analysed by liquid chromatography tandem mass spectrometry in order to measure the proteome of each fraction. In total, 811 individual proteins were identified, including 107, 84, and 146 proteins unique to superficial, deep-partial thickness and full-thickness burn wounds, respectively. The differences in the protein inventory and the associated gene ontologies represented within each burn depth category demonstrated that there are subtle, yet significant, variations in the biochemistry of burn wounds according to severity. Importantly, this study has produced the most comprehensive catalogue of proteins from the paediatric burn wound microenvironment to date. SIGNIFICANCE: To our knowledge, this study has been the first to comprehensively measure the paediatric burn blister fluid proteome and has provided insight into the proteomic response to burn injury. The study contributes to the knowledge of blister fluid biochemistry of burn injury and provides clinically relevant knowledge through the qualitative evaluation of biochemical differences between burns of different depths. A better understanding of the burn wound environment will ultimately assist with more accurate clinical decision making and improved wound healing and scar reduction procedures.

Mass spectrometry based data of the blister fluid proteome of paediatric burn patients.

The data presented here are associated with the article "The blister fluid proteome of paediatric burns" (Zang et al., 2016) [1]. Burn injury is a highly traumatic event for children. The degree of burn severity (superficial-, deep-, or full-thickness injury) often dictates the extent of later scar formation which may require long term surgical operation or skin grafting. The data were obtained by fractionating paediatric burn blister fluid samples, which were pooled according to burn depth and then analysed using data dependent acquisition LC-MS/MS. The data includes a table of all proteins identified, in which burn depth category they were found, the percentage sequence coverage for each protein and the number of high confidence peptide identifications for each protein. Further Gene Ontology enrichment analysis shows the significantly over-represented biological processes, molecular functions, and cellular components of the burn blister fluid proteome. In addition, tables include the proteins associated with the biological processes of "wound healing" and "response to stress" as examples of highly relevant processes that occur in burn wounds.

Evaluation of haemoglobin in blister fluid as an indicator of paediatric burn wound depth.

The early and accurate assessment of burns is essential to inform patient treatment regimens; however, this first critical step in clinical practice remains a challenge for specialist burns clinicians worldwide. In this regard, protein biomarkers are a potential adjunct diagnostic tool to assist experienced clinical judgement. Free circulating haemoglobin has previously shown some promise as an indicator of burn depth in a murine animal model. Using blister fluid collected from paediatric burn patients, haemoglobin abundance was measured using semi-quantitative Western blot and immunoassays. Although a trend was observed in which haemoglobin abundance increased with burn wound severity, several patient samples deviated significantly from this trend. Further, it was found that haemoglobin concentration decreased significantly when whole cells, cell debris and fibrinous matrix was removed from the blister fluid by centrifugation; although the relationship to depth was still present. Statistical analyses showed that haemoglobin abundance in the fluid was more strongly related to the time between injury and sample collection and the time taken for spontaneous re-epithelialisation. We hypothesise that prolonged exposure to the blister fluid microenvironment may result in an increased haemoglobin abundance due to erythrocyte lysis, and delayed wound healing.