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• W2037079144 abstract "Osteoarthritis is a chronic and progressive joint disease and it is considered the most prevalent form of arthritis. Its pathogenesis is believed to be resulting from an imbalance between processes of degradation and attempts of repair by chondrocytes. We propose unveiling the relationship of GRP with osteoarthritis, using 2D-DIGE. This protein was recently discovered by our research group and its presence is associated with calcium crystalsin situations of ectopic calcification, both in calcified arteries and skin calcinosis, which suggests that GRP in vertebrates may regulate calcium in the extracellular environment and also relates it to several pathologies associated with calcifications that appear at multiple locations of the organism like in osteoarthritis. The knowledge that GRP circulates systemically in the blood stream makes it a good candidate as a biomarker for ectopic calcification associated pathologies, and the correlation with results obtained from serum analysis, will provide insights into the mechanisms of cartilage degeneration/ calcification occurring in osteoarthritis. The first step of this work was the optimization of cartilage and synovial fluid extraction procedures and also of 2DE analysis conditions based on described studies of cartilage proteomic analysis and also in the knowledge about GRP specific chemical properties. The procedures were tested in order to obtain protein extracts containing predominantly extracellular matrix binding components including GRP. Following the extraction, samples were analyzed by 2DE and Western blot using the specific CTermGRP antibody. Our proteomic approach permitted not only the identification of GRP both in articular cartilage and synovial fluid but also enable us to detect and characterize all GRP forms present. Together, these results will stimulate and benefit research in themedicine area and increase our current understanding of the molecular mechanisms involved in cartilage degradation and calcification associated with osteoarthritis and more specifically the involvement of GRP in this processes. This work was partially supported by FCT funding (PTDC/SAU-ESA/101186/2008) and Sofia Cavaco is the recipient of a PhD grant from FCT (SFRH/BD/60867/2009). This article is part of a Special Issue entitled ECTS 2011. Disclosure of interest: None declared. doi:10.1016/j.bone.2011.03.652 PP522-T Effects of oxygen tension on the chondrogenic potential of differentially cultured human adipose-derived stem cells C. Merceron , S. Portron ⁎, M. Masson , J. Lesoeur , B.H. Fellah , O. Gauthier , P. Weiss , J. Guicheux , C. Vinatier a,c a INSERM U791 University of Nantes, France b ONIRIS Experimental Surgery Department, Nantes, France c Graftys SA, Aix en Provence, France Abstract: Cartilage has limited capacity for self repair. Biomaterial-assisted cell therapy using adult mesenchymal stem cells (MSC) has been considered promising for the treatment of cartilage Cartilage has limited capacity for self repair. Biomaterial-assisted cell therapy using adult mesenchymal stem cells (MSC) has been considered promising for the treatment of cartilage defect.Whilst bone-marrowderivedMSChas been extensively used for tissue engineering, adipose tissue has recently been contemplated owing to its accessibility and its MSC content. Articular cartilage is an avascular tissue, therefore, oxygen tension has been suggested as a regulatory factor for chondrogenic differentiation. In this context, our work aimed at deciphering the influence of oxygen tension on the in vitro and in vivo chondrogenic differentiation potential of human adiposederived stemcells (hATSC)when associatedwith a cellulose based self-setting hydrogel (Si-HPMC). hATSC isolated from lipoaspirates were characterized for their proliferation, surface markers expression and multipotency. To decipher the effects of hypoxia on the in vitro chondrogenic potential of hATSC, the cells were cultured in 2D or within Si-HPMC in control or chondrogenic medium and under normal (21%) or low (5%) oxygen tension. Chondrogenic differentiation was assessed by real time PCR for the expression of chondrogenic markers (COL2A1 and ACAN). To determine whether the in vitro differentiated hATSC allows cartilage formation in vivo, the cells induced in monolayer culture were associated with Si-HPMC hydrogel and implanted subcutaneously in nude mice for 5 weeks. Cartilage formation was evaluated by a histological scoring of nodule density as well as intensities of Alcian Blue staining and Type II collagen immunostaining (score from 0 to 5 for each criterion). Our data demonstrate that hATSC exhibited proliferation capability and expressed typical stem cell surface markers. In addition, they were able to differentiate towards the chondro-, osteoand adipogenic lineages. Real-time PCR analyses indicated that, in vitro, hypoxia induced an increased expression of chondrogenic markers as compared to normoxia. Besides, hATSC cultured in chondrogenic medium and whatever the oxygen tension exhibit ability to form a cartilaginous tissue in vivo as evidenced by the presence of sulphated GAG and type II collagen. These data could help us exploit the potential of stem cells to restore the function of degenerated cartilage by cell-based regenerative medicine. Acknowledgement: «Foundation arthritis Courtin», «Societe Francaise de Rhumatologie», ANR Tecsan and «Region Pays de La Loire» (Bioregos). This article is part of a Special Issue entitled ECTS 2011. Disclosure of interest: None declared. doi:10.1016/j.bone.2011.03.679 Abstracts / Bone 48 (2011) S261–S265 S265" @default.
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• W2037079144 date "2011-05-01" @default.
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• W2037079144 title "Identification and characterization of GRP in articular cartilage and synovial fluid using a proteomic-based approach" @default.
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