The proteome of blister fluid from paediatric burn injuries — ASN Events
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  3. The proteome of blister fluid from paediatric burn injuries

The proteome of blister fluid from paediatric burn injuries (#006)

Tuo Zang 1 2 , Daniel Broszczak 1 2 , James Broadbent 1 2 , Leila Cuttle 1 3 , Tony Parker 1 2
  1. TRR Program, IHBI-QUT, Kelvin Grove, Queensland, Australia
  2. School of Biomedical Sciences, Queensland University of Technology, Brisbane, Queensland, Australia
  3. Children’s Burns and Trauma Research, Centre for Children’s Health Research, South Brisbane, Queensland, Australia

Burn injury is highly traumatic for paediatric patients. Scar formation is a nature outcome in severe burn and dependent on the severity of the burn injury. The early and accurate diagnosis of medium to severe burn is different according to current clinical assessment which is mainly based on the clinician’s experience. These wounds represent a grey-area in scar formation and skin grafting. Thus, a more objective and quantitative measure to distinguish between burn severities is urgently required. Burn blister fluid is a viable study matrix that reflects systemic responses as well as the local microenvironment. This project seeks to characterise the blister fluid from children with different degrees of burn severity and determine severity specific protein markers to potentially aid in future clinical practice.

Blister fluid (BF) was collected from superficial thickness (n=12), deep-partial thickness (n=14), and full-thickness burn wounds (n=4). In order to characterise and identify BF proteins, the samples within each depth classification were pooled and subjected a variety of fractionation methods, including: filter aided sample preparation (FASP), lithium dodecyl sulphate polyacrylamide gel electrophoresis (LDS-PAGE), and Isoelectric OFFGel. Another 11 individual samples were processed by Multiple Affinity Removal Spin (MARS) cartridge. Then the enzymolytic products were analysed using LC-MS/MS to profile protein composition. The underlying biological processes were discovered using a gene ontology analysis.

We have demonstrated the strength of fractionation approaches to facilitate a deeper characterisation of burn blister fluid proteome. We identified, 228 proteins using the FASP, 365 by LDS-PAGE, 460 by MARS and 551 by OFFGel. Importantly, there were 812 different proteins identified in total including 107, 84, and 146 unique proteins in different burn severities, respectively, and 317 common to all three severities. The differences in proteins and biological processes demonstrated that there are subtle, yet significant, differences in the biochemistry of burn wounds according to severity. Moreover, this study represents the most comprehensive study of paediatric burn wound microenvironment to date.