Analysis of pressure ulcer wound fluid using two‐dimensional electrophoresis

Wyffels, Jennifer T.; Fries, Kristin M.; Randall, Jason S; Ha, Daniel S.; Lodwig, Christa A; Brogan, Michael S.; Shero, Marlene; Edsberg, Laura E.

doi:10.1111/j.1742-481x.2010.00672.x

Cited by 25 publications

(21 citation statements)

References 17 publications

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“…Previous work utilizing two‐dimensional polyacrylamide gel electrophoresis indicates a biochemical difference between the interior and the periphery of a wound as well as chronic and healed wounds. Protein S100A9 was identified by mass spectrometry of an excised gel spot as a potential biomarker of wound healing using this technique . As previously reported, this technique was not optimal for the characterization of the protein profile of chronic wounds and the alternate techniques reported in this paper were pursued.…”

Section: Discussionmentioning

confidence: 99%

“…The change in wound surface area expressed as a percentage of area on day 0 was used to place patients in one of three categories for wound outcome, chronic or not healed, moderate healing or healed. Collection and isolation of wound fluid proteins followed previously described protocols . In brief, polyester‐tipped applicators were rolled over surface of the wound interior and periphery independently until saturated.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of the proteomic profile of chronic pressure ulcers

Edsberg

Wyffels

Brogan

et al. 2012

Wound Repair Regeneration

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Analysis of the proteomic profile of pressure ulcers over time is a critical step in the identification of biomarkers of healing or nonhealing in pressure ulcers. The wound fluid from 32 subjects with 42 pressure ulcers was evaluated over 6 weeks at 15 time points. Samples specific to both the interior and the periphery of the wound bed were collected. Antibody screening arrays, isobaric tags for relative and absolute quantitation with mass spectrometry and multiplexed microarrays were used to characterize wound fluid and results were correlated with clinical outcome. Twenty-one proteins were found to distinguish between healed and chronic wounds and 19 proteins were differentially expressed between the interior and periphery of wounds. Four proteins, pyruvate kinase isozymes M1/M2, profilin-1, Ig lambda-1 chain C regions, and Ig gamma-1 chain C region, were present in lower levels for periphery samples when compared to interior samples and six proteins, keratin, type II cytoskeletal 6A (KRT6A), keratin, type I cytoskeletal 14, S100 calcium binding proteins A7, alpha-1-antitrypsin precursor, hemoglobin subunit alpha, and hemoglobin subunit beta, were present in higher levels in periphery samples when compared with interior samples. S100 calcium binding protein A6, S100 calcium binding protein A7, and soluble receptor for advanced glycation end-products had higher levels in the periphery of chronic wounds vs. the interior in planar arrays. A significant temporal trend was noted for monokine induced by gamma interferon (MIG), synonomous with chemokine (C-X-C motif) ligand 9 (CXCL9), which increased as wounds healed and remained nearly constant for ulcers that were not approaching closure.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Analysis of the proteomic profile of chronic pressure ulcers

Edsberg

Wyffels

Brogan

et al. 2012

Wound Repair Regeneration

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“…Alternatively, WF can be sampled discontinuously using wound swapping [19], occlusive dressings [20], porous dextranomer beads [21], capillary tubes, or paper strips [22, 23]. Using these methods the collected amount of protein is higher; however, the wound fluid contains salts as well as metabolites and the protein concentration is quite low and consequently requires additional cleaning and enrichment steps for proteomic analysis [17].…”

Section: Introductionmentioning

confidence: 99%

Proteomics and Metabolomics forIn SituMonitoring of Wound Healing

Kalkhof

Förster

Schmidt

et al. 2014

BioMed Research International

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Wound healing of soft tissue and bone defects is a complex process in which cellular differentiation and adaption are regulated by internal and external factors, among them are many different proteins. In contrast to insights into the significance of various single proteins based on model systems, the knowledge about the processes at the actual site of wound healing is still limited. This is caused by a general lack of methods that allow sampling of extracellular factors, metabolites, and proteins in situ. Sampling of wound fluids in combination with proteomics and metabolomics is one of the promising approaches to gain comprehensive and time resolved data on effector molecules. Here, we describe an approach to sample metabolites by microdialysis and to extract proteins simultaneously by adsorption. With this approach it is possible (i) to collect, enrich, and purify proteins for a comprehensive proteome analysis; (ii) to detect more than 600 proteins in different defects including more than 100 secreted proteins, of which many proteins have previously been demonstrated to have diagnostic or predictive power for the wound healing state; and (iii) to combine continuous sampling of cytokines and metabolites and discontinuous sampling of larger proteins to gain complementary information of the same defect.

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“…23 The stages of wound healing are distinct in function and require different proteins for their associated tasks. As ulcers progress through the stages of wound healing, tissue types in the wound change and concomitantly the protein profile of wound fluid.…”

Section: Discussionmentioning

confidence: 99%