Key regulatory molecules of cartilage destruction in rheumatoid arthritis: an in vitro study.

Arthritis Research & Therapy
Kristin AndreasMichael Sittinger

Abstract

Rheumatoid arthritis (RA) is a chronic, inflammatory and systemic autoimmune disease that leads to progressive cartilage destruction. Advances in the treatment of RA-related destruction of cartilage require profound insights into the molecular mechanisms involved in cartilage degradation. Until now, comprehensive data about the molecular RA-related dysfunction of chondrocytes have been limited. Hence, the objective of this study was to establish a standardized in vitro model to profile the key regulatory molecules of RA-related destruction of cartilage that are expressed by human chondrocytes. Human chondrocytes were cultured three-dimensionally for 14 days in alginate beads and subsequently stimulated for 48 hours with supernatants from SV40 T-antigen immortalized human synovial fibroblasts (SF) derived from a normal donor (NDSF) and from a patient with RA (RASF), respectively. To identify RA-related factors released from SF, supernatants of RASF and NDSF were analyzed with antibody-based protein membrane arrays. Stimulated cartilage-like cultures were used for subsequent gene expression profiling with oligonucleotide microarrays. Affymetrix GeneChip Operating Software and Robust Multi-array Analysis (RMA) were used to identif...Continue Reading

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Datasets Mentioned

BETA
GSE10024

Methods Mentioned

BETA
transgenic
in vitro transcription
protein membrane array
X-ray
protein array
enzyme-linked immunosorbent assay
transfection
PCR

Software Mentioned

GeneChip Operating Software ( GCOS )
Genesis
RMA
GeneChip Operating Software
GeneChip Operating Software Multi - array Analysis
GeneChip Operating
Robust Multi - array Analysis
GCOS
ABI Prism 777 Sequence Detection System

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