Dataset Information

Matrix-Bound Growth Factors are Released upon Cartilage Compression by an Aggrecan-Dependent Sodium Flux that is Lost in Osteoarthritis (Proteomics)

ABSTRACT: Articular cartilage exhibits regional and depth-dependent biomechanical differences, which influence the transduction of load to chondrocytes and throughout the tissue matrix. One such mechanism is the biophysical release of proteins from the matrix. This study conducted proteomic analysis of rested porcine articular cartilage to assess regional protein abundance throughout distinct depths of porcine knee articular cartilage.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Articular Cartilage

SUBMITTER: Patrik Onnerfjord

LAB HEAD: Patrik Onnerfjord

PROVIDER: PXD023890 | Pride | 2021-07-23

REPOSITORIES: Pride

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			Action	DRS
	160226_TV_cntrl_bov_1.raw	Raw
	160226_TV_cntrl_bov_10.raw	Raw
	160226_TV_cntrl_bov_11.raw	Raw
	160226_TV_cntrl_bov_12b.raw	Raw
	160226_TV_cntrl_bov_2.raw	Raw

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Project description:The purpose of this study was to characterize the histologic development of OA in a mouse model where OA is induced by destabilization of the medial meniscus (DMM model) and to identify genes regulated during different stages of the disease, using RNA isolated from the joint M-bM-^@M-^\organM-bM-^@M-^] and analyzed using microarrays.427 genes from the microarrays passed consistency and significance filters. There was an initial up-regulation at 2 and 4 weeks of genes involved in morphogenesis, differentiation, and development, including growth factor and matrix genes, as well as transcription factors including Atf2, Creb3l1, and Erg. Most genes were off or down-regulated at 8 weeks with the most highly down-regulated genes involved in cell division and the cytoskeleton. Gene expression increased at 16 weeks, in particular extracellular matrix genes including Prelp, Col3a1 and fibromodulin.The results support a phasic development of OA with early matrix remodelling and transcriptional activity followed by a more quiescent period that is not maintained. A group of 9 mice was used for collection of RNA at time 0 (before surgery) when the animals were 12 weeks old. For the other time points, 9 DMM and 9 sham controls were sacrificed at 2, 4, 8, and 16 weeks after surgery for RNA isolation. The tissue included tibial plateau and femoral condyle articular cartilage, subchondral bone with any osteophytes, meniscus, and the joint capsule with synovium was used for RNA isolation. The tissue was treated with RNAlaterM-BM-. (Invitrogen) prior to freezing and storage at -800 C. RNA was extracted by homogenization using the Precellys 24 tissue homogenizer (Bertin Technologies purchased from MO BIO) and the amount and quality of the RNA was determined using an Agilent 2100 Bioanalyzer. RNA was pooled prior to microarray analysis such that 3 randomly selected samples from each surgical group and time point were pooled to create each biological replicate. Because 9 mice were used for each experimental group, a total of three biological replicates per group were analyzed using the Affymetrix Mouse Genome 430 2.0 oligonucleotide arrays as described. One replicate pool, which was from week two DMM mice, did not meet the RNA integrity level needed for microarray analysis; thus, this pool was not analyzed further, leaving two pools for the week two DMM mice.

Dataset Information

Matrix-Bound Growth Factors are Released upon Cartilage Compression by an Aggrecan-Dependent Sodium Flux that is Lost in Osteoarthritis (Proteomics)

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