Characterization of neutrophil chemoattractants in inflamed synovial fluid Method development and evaluation for the study of inflamed synovial fluid from patients with inflammatory arthritis

Abstract

Inflammatory arthritis is a group of inflammatory diseases mainly characterized by inflammation in synovial cavities, eventually leading to the destruction of cartilage and bone in joints. The complete mechanisms behind this inflammatory process are not fully understood today, but some signs indicate that it might differ from the nor mal inflammatory response associated with acute inflammation. This project focuses on signaling molecules, called chemoattractants, that are responsible for recruiting neutrophils to the inflamed synovial fluid in patients with inflammatory arthritis. More specifically, the aim is to develop and evaluate a possible methodology as a first step toward identifying and characterizing the neutrophil chemoattractants. In order to do this, the complexity of the problem can be reduced by dividing the com ponents of synovial fluid into fractions based on different molecular properties. Two possible approaches were investigated here; one where the components were divided through gel electrophoresis, and one where the components were separated using liq uid chromatography methods, after which their chemotactic abilities were measured with a chemotaxis assay. The separated fractions yielded from both approaches were then measured with liquid chromatography-mass spectrometry to obtain more information and clues about the contents and success of separations. Based on the results, the course of action based on liquid chromatography and chemotaxis assay was concluded as most promising for successful separation of components and nar rowing of the problem, as is proposed as a first step of methodology in further search for the neutrophil chemoattractants.