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Periostin expression in inflammatory and non-inflammatory osteoarthritis and rheumatoid arthritis
Abstracts / Osteoarthritis and Cartilage 24 (2016) S63eS534
S69
102 PERIOSTIN EXPRESSION IN INFLAMMATORY AND NONINFLAMMATORY OSTEOARTHRITIS AND RHEUMATOID ARTHRITIS N. Athanasou y, A. Mantoku z, A. Kudo z, Y. Inagaki y, D. Mahoney y, T. Kashima y. y Oxford Univ., Oxford, United Kingdom; z Dept. of Biological Information, Tokyo Inst. of Technology, Yokohama, Japan
Expression stability values of the candidate reference genes calculated by Normfinder Symbol
Stability
YWHAZ ACTB RPL13A B2M HPRT1 GAPDH SDHA
0.1 0.18 0.2 0.21 0.22 0.24 0.25
Coefficient of correlation (r) of the candidate reference genes calculated by BestKeeper Symbol
r
YWHAZ ACTB SDHA RPL13A B2M HPRT1 GAPD
0.989 0.983 0.979 0.975 0.974 0.961 0.945
Purpose: Periostin is a secreted extracellular matrix protein which belongs to the fasciclin family. It plays a role in embryogenesis and tissue repair and shows increased expression in tissues subject to mechanical stress and in some neoplastic and inflammatory conditions. Periostin has been identified in synovium and cartilage in osteoarthritis (OA) and in inflammatory arthritis synovium. To determine the role of periostin in inflammatory OA and non-inflammatory OA we characterized periostin expression in the synovium and synovial fluid in cases of non-inflammatory and non-inflammatory OA and in normal and rheumatoid arthritis (RA) synovium. Methods: Synovial fluid (SF) was aspirated from the knee joints of 9 patients with OA and 9 patients with (RA). Periostin levels were assessed by ELISA. Immunohistochemistry was carried out by an indirect immuno-peroxidase technique on 60 samples of knee joint synovium from RA and non-inflammatory and inflammatory OA cases as well as controls of normal synovium using a polyclonal antibody raised against human periostin. Results: The average periostin concentration in synovial fluid was 107.4mg/ml and 67.1 mg/ml in RA and OA patients respectively. Immunohistochemistry showed strong expression of periostin in RA and inflammatory OA synovial tissues, particularly in the extracellular matrix of the superficial subintima beneath the synovial lining; this was mostly related to increased vascularity in this zone. Most cases of non-inflammatory OA showed relatively little expression of periostin in this area. Periostin was expressed in the matrix around small and large vessels in the deeper subintima of OA and RA cases as well as in areas of degenerative change within the synovium and joint capsule. Conclusions: Our findings indicate that increased levels of periostin are seen in the context of a chronic inflammatory synovitis; in both RA and inflammatory OA; there was markedly less expression of periostin in non-inflammatory OA synovium. Increased periostin expression in inflamed synovium is mirrored by the observation that the level of periostin is increased in RA compared with OA synovial fluid. Periostin is expressed by fibroblast-like cells in the synovium and it has been identified in OA cartilage and synovium. Increased periostin expression in inflammatory OA may be related to increased synovial fibroblast activity in inflamed synovial tissue. It has been suggested that periostin could be employed as a serum biomarker for OA. Our results suggest that serum levels of periostin are likely to be higher in inflammatory compared with non-inflammatory OA and that periostin may represent an immunophenotypic marker for distinguishing inflammatory from non-inflammatory OA.
Biomarker 103 ESTABLISHMENT OF A CO-CULTURE SYSTEM PROGRESSION WITHOUT ADDING OF CYTOKINES
TO
MIMIC
OA
Standard deviation (SD) of the candidate reference genes calculated by BestKeeper
T. Mang y, D. Werkmann y, A.C. Bay-Jensen z, A. Siebuhr z, M.A. Karsdal z, C.H. Ladel y, A. Gigout y, H. Guehring y. y Merck KGaA, Darmstadt, Germany; z Nordic BioSci., Herlev, Denmark
Symbol
SD
RPL13A HPRT1 GAPD YWHAZ B2M SDHA ACTB
0.61 0.69 0.74 0.79 0.84 0.92 0.95
Purpose: Osteoarthritis (OA) is a highly complex degenerative joint disease, which involves all joint tissues. Several ambitious attempts have been made to develop disease-modifying treatments (DMOADs), but all have failed so far. The use of artificial in-vitro test systems may contribute to unravel reasons of failures. Knowing that OA involves not only cartilage, we investigated the effect of co-culturing various joint tissues with cartilage in order to allow autocrine and paracrine interplay of these tissues. The aim of this study was to identify a culture condition in which cartilage degradation occur without adding of external cytokines.
S69
102 PERIOSTIN EXPRESSION IN INFLAMMATORY AND NONINFLAMMATORY OSTEOARTHRITIS AND RHEUMATOID ARTHRITIS N. Athanasou y, A. Mantoku z, A. Kudo z, Y. Inagaki y, D. Mahoney y, T. Kashima y. y Oxford Univ., Oxford, United Kingdom; z Dept. of Biological Information, Tokyo Inst. of Technology, Yokohama, Japan
Expression stability values of the candidate reference genes calculated by Normfinder Symbol
Stability
YWHAZ ACTB RPL13A B2M HPRT1 GAPDH SDHA
0.1 0.18 0.2 0.21 0.22 0.24 0.25
Coefficient of correlation (r) of the candidate reference genes calculated by BestKeeper Symbol
r
YWHAZ ACTB SDHA RPL13A B2M HPRT1 GAPD
0.989 0.983 0.979 0.975 0.974 0.961 0.945
Purpose: Periostin is a secreted extracellular matrix protein which belongs to the fasciclin family. It plays a role in embryogenesis and tissue repair and shows increased expression in tissues subject to mechanical stress and in some neoplastic and inflammatory conditions. Periostin has been identified in synovium and cartilage in osteoarthritis (OA) and in inflammatory arthritis synovium. To determine the role of periostin in inflammatory OA and non-inflammatory OA we characterized periostin expression in the synovium and synovial fluid in cases of non-inflammatory and non-inflammatory OA and in normal and rheumatoid arthritis (RA) synovium. Methods: Synovial fluid (SF) was aspirated from the knee joints of 9 patients with OA and 9 patients with (RA). Periostin levels were assessed by ELISA. Immunohistochemistry was carried out by an indirect immuno-peroxidase technique on 60 samples of knee joint synovium from RA and non-inflammatory and inflammatory OA cases as well as controls of normal synovium using a polyclonal antibody raised against human periostin. Results: The average periostin concentration in synovial fluid was 107.4mg/ml and 67.1 mg/ml in RA and OA patients respectively. Immunohistochemistry showed strong expression of periostin in RA and inflammatory OA synovial tissues, particularly in the extracellular matrix of the superficial subintima beneath the synovial lining; this was mostly related to increased vascularity in this zone. Most cases of non-inflammatory OA showed relatively little expression of periostin in this area. Periostin was expressed in the matrix around small and large vessels in the deeper subintima of OA and RA cases as well as in areas of degenerative change within the synovium and joint capsule. Conclusions: Our findings indicate that increased levels of periostin are seen in the context of a chronic inflammatory synovitis; in both RA and inflammatory OA; there was markedly less expression of periostin in non-inflammatory OA synovium. Increased periostin expression in inflamed synovium is mirrored by the observation that the level of periostin is increased in RA compared with OA synovial fluid. Periostin is expressed by fibroblast-like cells in the synovium and it has been identified in OA cartilage and synovium. Increased periostin expression in inflammatory OA may be related to increased synovial fibroblast activity in inflamed synovial tissue. It has been suggested that periostin could be employed as a serum biomarker for OA. Our results suggest that serum levels of periostin are likely to be higher in inflammatory compared with non-inflammatory OA and that periostin may represent an immunophenotypic marker for distinguishing inflammatory from non-inflammatory OA.
Biomarker 103 ESTABLISHMENT OF A CO-CULTURE SYSTEM PROGRESSION WITHOUT ADDING OF CYTOKINES
TO
MIMIC
OA
Standard deviation (SD) of the candidate reference genes calculated by BestKeeper
T. Mang y, D. Werkmann y, A.C. Bay-Jensen z, A. Siebuhr z, M.A. Karsdal z, C.H. Ladel y, A. Gigout y, H. Guehring y. y Merck KGaA, Darmstadt, Germany; z Nordic BioSci., Herlev, Denmark
Symbol
SD
RPL13A HPRT1 GAPD YWHAZ B2M SDHA ACTB
0.61 0.69 0.74 0.79 0.84 0.92 0.95
Purpose: Osteoarthritis (OA) is a highly complex degenerative joint disease, which involves all joint tissues. Several ambitious attempts have been made to develop disease-modifying treatments (DMOADs), but all have failed so far. The use of artificial in-vitro test systems may contribute to unravel reasons of failures. Knowing that OA involves not only cartilage, we investigated the effect of co-culturing various joint tissues with cartilage in order to allow autocrine and paracrine interplay of these tissues. The aim of this study was to identify a culture condition in which cartilage degradation occur without adding of external cytokines.