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Extracellular mitochondrial DNA and oxidatively damaged DNA in synovial fluid of patients with rheumatoid arthritis.doc

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Extracellular mitochondrial DNA and oxidatively damaged DNA in synovial fluid of patients with rheumatoid arthritis

Arthritis Research Therapy Vol 5 No 5 Hajizadeh et al. Open Access Research article Extracellular mitochondrial DNA and oxidatively damaged DNA in synovial fluid of patients with rheumatoid arthritis Shahin Hajizadeh1, Jeroen DeGroot2, Johan M TeKoppele2, Andrej Tarkowski1 and L Vincent Collins1 1Department of Rheumatology and Inflammation Research, University of G?teborg, G?teborg, Sweden 2TNO Prevention and Health, Gaubius Laboratory, Leiden, The Netherlands Corresponding author: Shahin Hajizadeh (e-mail: shahin@rheuma.gu.se) Received: 21 Mar 2003 Revisions requested: 29 Apr 2003 Revisions received: 15 May 2003 Accepted: 27 May 2003 Published: 25 Jun 2003 Arthritis Res Ther 2003, 5:R234-R240 (DOI 10.1186/ar787) ? 2003 Hajizadeh et al., licensee BioMed Central Ltd (Print ISSN 1478-6354; Online ISSN 1478-6362). This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the articles original URL. Abstract We investigated whether plasma and synovial fluid (SF) samples from patients with rheumatoid arthritis (RA) contained extracellular mito- chondrial DNA (mtDNA) or the oxidatively damaged DNA adduct 8-hydroxy-2′-deoxyguanosine (8-oxodG). Moreover, we correlated the laboratory findings of the patients with RA with their levels of mtDNA and 8-oxodG. SF and plasma samples from 54 patients with RA, SF from 30 non-arthritic control subjects, and plasma from 22 healthy vol- unteers were collected. The samples were subjected to polymerase chain reaction (PCR) using mitochondrial genomic primers, and the products were analyzed by SDS–polyacrylamide-gel electrophoresis. The intensities of the PCR-amplified bands were quantified and normal- ized to a reference sample. Furthermore, the SF samples were assayed by enzyme-linked immunosorbent assay for 8-oxodG. Extracellular PCR-amplifiable mtDNA w