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Next Generation Sequencing of Transcriptomic microRNAs from Murine Articular Chondrocytes Treated with IL-1 [microRNA-seq]

ABSTRACT: Murine articular chondrocytes were isolated as previously described (Gosset et al., 2008) with modifications. Briefly, articular cartilages were dissected from femoral heads of 3-week-old wild-type C57/BL6J mice and digested for 4-6 hours in 0.5 mg/mL collagenase P (Roche, # 11249002001) in high-glucose DMEM (Thermo Fisher Scientific, #10569010) supplemented with 1% penicillin/streptomycin. Following digestion, cells were collected and seeded at a density of 50 x 104 cells/well in 12-well plates. The next day, cells were treated with IL-1 (1 ng/mL) (R&D Systems, #201-LB-005) for 24 hours. Total RNA were then extracted from cell cultures and assayed for microRNA sequencing.

ORGANISM(S): Mus musculus

PROVIDER: GSE221484 | GEO | 2023/02/06

REPOSITORIES: GEO

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Project description:Background: Meniscus tears are the most common injury in the knee and are associated with an increased risk of osteoarthritis (OA). The molecular profile of knees with meniscus tears is not well-studied. Therefore, to advance our understanding of the early response of the knee to injury, we compared the gene expression profile of meniscus and articular cartilage within the same knees following meniscus injury. Hypothesis/Purpose: To identify differences between the molecular signatures of meniscus and articular cartilage from knees with intact articular cartilage undergoing arthroscopic partial meniscectomy. Study Design: Descriptive laboratory study Methods: Patients (n=12) with a known isolated medial meniscus tear without any knee chondrosis or radiographic OA were consented prior to surgery. During arthroscopic partial meniscectomy, a sample of their injured meniscus and a sample of their articular cartilage off the medial femoral condyle were procured. The transcriptome signatures, as measured through Affymetrix microarray, were compared between the two tissues and underlying biological processes were explored computationally. Results: 3566 gene transcripts were differentially expressed between meniscus and articular cartilage. Gene transcripts down-regulated in articular cartilage were associated with extracellular matrix organization, wound healing, cell adhesion, and chemotaxis. Gene transcripts up-regulated in articular cartilage were associated with blood vessels morphogenesis and angiogenesis. Examples of individual genes with significant differences in expression between the two tissues include IBSP (23.76 fold; P < 0.001), upregulated in meniscus, and TREM1 (3.23 fold; P = 0.006), upregulated in meniscus. Conclusion: The meniscus and articular cartilage have distinct gene expression profiles in knees with meniscus tears and intact articular cartilage. Total RNA obtained from injured meniscus and normal articular cartilage from patients undergoing partial meniscectomy.

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Next Generation Sequencing of Transcriptomic microRNAs from Murine Articular Chondrocytes Treated with IL-1 [microRNA-seq]

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