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PLoS One. 2016 Jul 21;11(7):e0159580. doi: 10.1371/journal.pone.0159580. eCollection 2016.

Microdialysis Sampling from Wound Fluids Enables Quantitative Assessment of Cytokines, Proteins, and Metabolites Reveals Bone Defect-Specific Molecular Profiles.

Author information

1
University Center of Orthopedics and Trauma Surgery and Center for Translational Bone, Joint and Soft Tissue Research, University Hospital "Carl Gustav Carus", TU Dresden, Dresden, Germany.
2
Department of Molecular Systems Biology, Helmholtz-Centre for Environmental Research - UFZ, Leipzig, Germany.
3
Institute of Pharmacy, Faculty of Biosciences, Pharmacy and Psychology, University of Leipzig, Leipzig, Germany.
4
Division of Endocrinology and Metabolic Bone Diseases, Department of Medicine III, University Hospital "Carl Gustav Carus", TU Dresden, Dresden, Germany.
5
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, University of Leipzig, Leipzig, Germany.
6
Centre for Microbial Communities, University of Aalborg, Aalborg East, Denmark.
7
Department of Bioanalytics, University of Applied Sciences and Arts of Coburg, Coburg, Germany.
8
Center for Regenerative Therapies Dresden (CRTD), Dresden, Germany.

Abstract

Bone healing involves a variety of different cell types and biological processes. Although certain key molecules have been identified, the molecular interactions of the healing progress are not completely understood. Moreover, a clinical routine for predicting the quality of bone healing after a fracture in an early phase is missing. This is mainly due to a lack of techniques to comprehensively screen for cytokines, growth factors and metabolites at their local site of action. Since all soluble molecules of interest are present in the fracture hematoma, its in-depth assessment could reveal potential markers for the monitoring of bone healing. Here, we describe an approach for sampling and quantification of cytokines and metabolites by using microdialysis, combined with solid phase extractions of proteins from wound fluids. By using a control group with an isolated soft tissue wound, we could reveal several bone defect-specific molecular features. In bone defect dialysates the neutrophil chemoattractants CXCL1, CXCL2 and CXCL3 were quantified with either a higher or earlier response compared to dialysate from soft tissue wound. Moreover, by analyzing downstream adaptions of the cells on protein level and focusing on early immune response, several proteins involved in the immune cell migration and activity could be identified to be specific for the bone defect group, e.g. immune modulators, proteases and their corresponding inhibitors. Additionally, the metabolite screening revealed different profiles between the bone defect group and the control group. In summary, we identified potential biomarkers to indicate imbalanced healing progress on all levels of analysis.

PMID:
27441377
PMCID:
PMC4956113
DOI:
10.1371/journal.pone.0159580
[Indexed for MEDLINE]
Free PMC Article

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