Dataset Information

Equine OA model - An inflammatory equine model demonstrates dynamic changes of immune response and cartilage matrix molecule degradation in vitro

ABSTRACT: Articular cartilage explants were incubated with or without interleukin-1β for 25 days. Media samples collected at 3-day intervals were analysed by quantitative proteomics. Semi-tryptic COMP peptides containing novel neo-epitopes were identified and quantified in cell culture media from cartilage explants.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Cell Culture

SUBMITTER: Ulla Rüetschi

LAB HEAD: Ulla Ruetschi

PROVIDER: PXD004993 | Pride | 2018-01-05

REPOSITORIES: Pride

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An inflammatory equine model demonstrates dynamic changes of immune response and cartilage matrix molecule degradation in vitro.

Svala Emilia E Löfgren Maria M Sihlbom Carina C Rüetschi Ulla U Lindahl Anders A Ekman Stina S Skiöldebrand Eva E

Connective tissue research 20150422 4

The molecular aspects of inflammation were investigated in equine articular cartilage explants using quantitative proteomics. Articular cartilage explants were stimulated with interleukin (IL)-1β in vitro for 25 days, and proteins released into cell culture media were chemically labeled with isobaric mass tags and analyzed by liquid chromatography-tandem mass spectrometry. A total of 127 proteins were identified and quantified in media from explants. IL-1β-stimulation resulted in an abundance of ...[more]

PMID: 25803623

Publication: 1/2

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Project description:Osteoarthritis (OA) is an age-related degenerative musculoskeletal disease characterised by loss of articular cartilage, synovitis, abnormal bone proliferation and subchondral bone sclerosis. The underlying pathogenesis of OA is yet to be fully elucidated with no OA specific biomarkers in clinical use. Nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) allow identification of the global metabolome and proteome respectively. During this study, ex-vivo equine cartilage explants (n=5) were incubated in TNF-α/IL-1β supplemented culture media for 8 days, with media removed and replaced at 2, 5 and 8 days. Acetonitrile metabolite extractions of 8 day cartilage explants and media samples at all time points underwent 1H NMR metabolic analysis with media samples also undergoing MS proteomic analysis. Within the cartilage, metabolites glucose and lysine were elevated following TNF-α/IL-1β treatment whilst adenosine, alanine, betaine, creatine, myo-inositol and uridine levels decreased. Within the culture media, four, four and six metabolites were identified as being differentially abundant between control and treatment groups for 1-2 day, 3-5 day and 6-8 day time points respectively. Culture media proteomics identified 154, 138 and 72 proteins differentially abundant, with > 2 fold change, between control and treatment groups for 1-2 day, 3-5 day and 6-8 day time points respectively. Nine potential novel OA neopeptides were elevated in treated media. This is the first study to use a multi ‘omics’ approach to simultaneously investigate the metabolomic profile of ex-vivo cartilage and metabolomic/proteomic profiles of culture media using the TNF-α/IL-1β ex-vivo OA cartilage model. This study has identified a panel of metabolites, proteins and extracellular matrix derived neopeptides which are differentially abundant during an early phase of the OA model which may provide further information on underlying disease pathogenesis, allow potential translation for clinical markers and possible novel therapeutic targets.

			Action	DRS
	Experimentallayout.pdf	Pdf
	F020253_pep_set1.xml	Xml
	F020257_pep_set2.xml	Xml
	F020261_pep_set3.xml	Xml
	Set1.mgf	Mgf