Analysis of the proteomic profile of chronic pressure ulcers.

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.

Potential biomarkers of temporomandibular joint disorders.

PURPOSE: The purpose of this study was to identify protein markers present in subjects with temporomandibular joint disorders (TMDs) and clicking compared with the levels in controls. MATERIALS AND METHODS: This was a pilot case-control study, and we report the preliminary results. Samples of joint aspirate collected from patients with TMDs and controls who had undergone surgery for a problem other than TMDs were analyzed using isobaric tags for relative and absolute quantitation (iTRAQ) and biotin-labeled-based protein arrays. The data obtained from these techniques were used to identify the proteins of interest, which were then quantitated using enzyme-linked immunosorbent assay (ELISA). The patient samples studied included joint aspirate collected clinically from the controls and patients and included samples from both the right and the left sides of each patient with a TMD. RESULTS: The 8 TMJ aspirate samples from 6 subjects included 5 aspirate samples from 4 patients and 3 from 2 controls. The greatest standardized protein concentration of endocrine gland-derived vascular endothelial growth factor/prokineticin-1 (EG-VEGF/PK1) and D6 was found in both joints of the controls compared with the levels from the joints of the patients. With 1 exception, the standardized protein concentration was significantly lower in the patients than in the controls. The lower levels of EG-VEGF/PK1 and D6 in the patients compared with the controls suggest that these cytokines might be possible biomarkers for TMDs. CONCLUSION: In the present pilot study, greater levels of EG-VEGF/PK1 and D6 were found in the controls than in the patients with TMDs. Proteomic analysis of the proteins present in the diseased joints compared with those in the controls might help to identify proteins present when pain or degeneration of the joint occurs. The proteomic information might be useful in the development of future therapies.

Analysis of pressure ulcer wound fluid using two-dimensional electrophoresis.

The incidence rate of pressure ulcers in the USA ranges from 0.4% to 38% in acute care settings and from 2.2% to 23.9% in long-term care settings, and their treatment costs are in the billions of dollars yearly. The proteome of wound fluid may contain early indicators or biomarkers associated with healing in pressure ulcers that would enable treatment regimes to be optimised for each individual. Wound fluid was collected from the interior and periphery of 19 chronic pressure ulcers at 15 time points during 42 days for an analysis of protein expression. Proteins were fractionated using two-dimensional polyacrylamide gel electrophoresis. A comparison of the spot distributions indicates a biochemical difference between the interior and the periphery of wounds. Pressure ulcers that healed show a greater number of spots for interior and peripheral locations combined over time when compared with wounds that did not heal. Using this technique, protein S100A9 was identified as a potential biomarker of wound healing. The identification of differences within the proteome of healing versus non healing pressure ulcers could have great significance in the use of current treatments, as well as the development of new therapeutic interventions.

Analysis of the collagen I and fibronectin of temporomandibular joint synovial fluid and discs.

PURPOSE: The objective of this study was to investigate the content of synovial fluid aspirates and temporomandibular joint (TMJ) disc tissue for collagen I and total fibronectin in patients with closed lock. Fibronectin contains dual properties of assisting with wound healing and inducing cartilage degradation. Native fibronectin has been shown to assist with wound repair, whereas particular fibronectin fragments may degrade cartilage. In addition, collagen I is the major supporting protein of the TMJ disc and will degrade as osteoarthritis progresses. Fibronectin or collagen I expression in human TMJ synovial aspirates and disc tissue may indicate the proteins' involvement in closed lock. The hypothesis of this study is that TMJ discs and serum of patients with closed lock will contain an increased amount of fibronectin and decreased amount of collagen I. MATERIALS AND METHODS: We analyzed a total of 8 diseased TMJ discs and 4 diseased synovial fluid aspirates. For our control samples, we assessed 5 synovial samples from healthy patients and control skin samples. Using an enzyme-linked immunosorbent assay allowed us to measure the total amount of fibronectin and collagen I in synovial aspirates. Furthermore, we used light microscopy to assess TMJ disc histology and collagen architecture in control skin samples. Lastly, using fluorescent staining, we examined fibronectin and collagen I expression in TMJ discs. We compared the fluorescent staining and light microscopy results of both proteins within each disc to confirm fibronectin and collagen I expression. RESULTS: Disc specimens with advanced morphologic pathology showed significant labeling for fibronectin in 3 of 3 cases and for collagen I in 4 of 4 cases. There was no considerable difference in detection of either fibronectin or collagen I in TMJ synovial aspirates from patients with advanced disc pathology compared with controls. CONCLUSIONS: The levels of fibronectin and collagen I in the TMJ disc and synovial fluid may be influenced by the stage of disease. The results did not provide a clear understanding of fibronectin and collagen I involvement with tissue repair in closed-lock cases. Detection of fibronectin fragments may provide more meaningful results.