Biomarkers of inflammation and innate immunity in atrophic nonunion fracture

Journal of Translational Medicine

Research

Pierre Leprince¹  Marianne Fillet²  Myrielle Mathieu³  Valérie Gangji⁴  Charlotte Collin⁵  Dominique de Seny⁵  Jean-Philippe Hauzeur⁵  Michel G. Malaise⁵  Gaël Cobraiville⁶ 

[1] GIGA-Neurosciences, University of Liège, 4000, Liège, Belgium;Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liège, 4000, Liège, Belgium;Laboratory of Bone and Metabolic Biochemistry, Department of Rheumatology, Université Libre de Bruxelles (ULB), 1000, Brussels, Belgium;Laboratory of Bone and Metabolic Biochemistry, Department of Rheumatology, Université Libre de Bruxelles (ULB), 1000, Brussels, Belgium;Department of Rheumatology and Physical Medicine, Hôpital Erasme, Université Libre de Bruxelles (ULB), 1000, Brussels, Belgium;Laboratory of Rheumatology, Department of Rheumatology, GIGA Research, University of Liège, Tour GIGA, +2, CHU, 4000, Liège, Belgium;Laboratory of Rheumatology, Department of Rheumatology, GIGA Research, University of Liège, Tour GIGA, +2, CHU, 4000, Liège, Belgium;Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liège, 4000, Liège, Belgium;

关键词: Proteomics;    Biomarkers;    2D-DIGE;    SELDI;    Nonunion;    Innate immunity;    Hepcidin;    Complement;    Apolipoprotein;    S100A8;   

DOI  :  10.1186/s12967-016-1019-1

 received in 2016-04-26, accepted in 2016-08-22,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundNonunion is a failure of healing following a bone fracture. Its physiopathology remains partially unclear and the discovery of new mediators could promote the understanding of bone healing.MethodsThirty-three atrophic nonunion (NU) patients that failed to demonstrate any radiographic improvement for 6 consecutive months were recruited for providing serum samples. Thirty-five healthy volunteers (HV) served as the control group. Proteomics studies were performed using SELDI-TOF–MS and 2D-DIGE approaches, associated or not with Proteominer® preprocessing, to highlight biomarkers specific to atrophic nonunion pathology. Peak intensities were analyzed by two statistical approaches, a nonparametric Mann–Whitney U tests (univariate approach) and a machine-learning algorithm called extra-trees (multivariate approach). Validation of highlighted biomarkers was performed by alternative approaches such as microfluidic LC–MS/MS, nephelometry, western blotting or ELISA assays.ResultsFrom the 35 HV and 33 NU crude serum samples and Proteominer® eluates, 136 spectra were collected by SELDI-TOF–MS using CM10 and IMAC-Cu2+ ProteinChip arrays, and 665 peaks were integrated for extra-trees multivariate analysis. Accordingly, seven biomarkers and several variants were identified as potential NU biomarkers. Their levels of expression were found to be down- or up-regulated in serum of HV vs NU. These biomarkers are inter-α-trypsin inhibitor H4, hepcidin, S100A8, S100A9, glycated hemoglobin β subunit, PACAP related peptide, complement C3 α-chain. 2D-DIGE experiment allowed to detect 14 biomarkers as being down- or up-regulated in serum of HV vs NU including a cleaved fragment of apolipoprotein A-IV, apolipoprotein E, complement C3 and C6. Several biomarkers such as hepcidin, complement C6, S100A9, apolipoprotein E, complement C3 and C4 were confirmed by an alternative approach as being up-regulated in serum of NU patients compared to HV controls.ConclusionTwo proteomics approaches were used to identify new biomarkers up- or down-regulated in the nonunion pathology, which are involved in bone turn-over, inflammation, innate immunity, glycation and lipid metabolisms. High expression of hepcidin or S100A8/S100A9 by myeloid cells and the presence of advanced glycation end products and complement factors could be the result of a longstanding inflammatory process. Blocking macrophage activation and/or TLR4 receptor could accelerate healing of fractured bone in at-risk patients.

【 授权许可】

CC BY   
© The Author(s) 2016