Detection of bone and cartilage-related proteins in plasma of patients with a bone fracture using liquid chromatography–mass spectrometry

Abstract

Following bone fracture, a large number of growth factors, cytokines, and their cognate receptors involved in the repair process are active at the fracture site. To determine whether they appear in patients' blood as candidate biomarkers for following the outcome of healing, we analysed the plasma of 25 patients with an acute bone fracture following affinity plasma purification, SDS gel electrophoresis and liquid chromatography -tandem mass spectrometry (LC-MS/ MS). Two hundred and thirteen nonredundant proteins were identified in the in-gel analysis of pooled plasma proteins. Gene ontology (GO) analysis indicated that a majority of detected proteins were of extracellular origin, whereas only a small number were of intracellular (cytosol and nucleus) origin. A significant proportion of detected proteins was involved in the cell growth and proliferation, transport and coagulation. Twelve proteins were potentially related to bone and cartilage metabolism, and several have not been previously identified in the plasma, including: TGF-β induced protein IG-H 3 , cartilage acidic protein 1, procollagen C proteinase enhancer protein and TGF-β receptor III.Résumé Après une fracture, un grand nombre de facteurs de croissance, cytokines et leurs récepteurs apparentés interviennent dans le processus de réparation des foyers de fracture. Nous avons analysé ces différents facteurs circulants chez 25 patients ayant présenté une fracture après purification du sang, électrophorèses, chromatographie et spectrographie de masse. 213 protéines ont été identifiées. L'analyse génétique de la majorité de ces protéines montre qu'elles sont d'origine extra cellulaires avec un très petit nombre de protéines intra cellulaires provenant notamment du noyau. Une proportion significative des protéines détectées intervient au niveau de la croissance, de la prolifération cellulaire et des phénomènes de coagulation. 12 protéines sont spécifiquement en rapport avec les métabolismes osseux et cartilagineux, plusieurs d'entre-elles n'avaient pas été préala-blement identifiées au niveau du plasma comme la TGF-β, la protéine IG-H3, la CAP 1, le procollagène de type C, le TGF-β récepteur III.

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