Project description:Effect of stimulation with IL-1beta and p38 MAPK inhibition with SB203580 or Birb 796 on human articular osteoarthritic chondrocytes
Project description:This study used high-throughput RNA sequencing to investigate the combination of IL-1b stimulation and IL-1b PGG-Zn@Cur Transcriptome profile of chondrocytes under pH Responsive Zn ² ⁺ - Coordinated Gellan Gum/γ - Polyglutamic Acid loaded with Curculigoside. Mouse primary chondrocytes were isolated from the articular cartilage of mouse knee joints and cultured under corresponding treatment conditions. Extract total RNA and perform paired end RNA seq on the Illumina NovaSeq 6000 platform. Use HISAT2 to process raw sequencing readings and compare them with GRCh38. The gene expression level was quantified as TPM and differentially expressed genes were identified using DESeq2. This dataset provides comprehensive transcriptomic insights into the molecular mechanisms of chondrocyte biology in the context of disease models.
Project description:Bovine articular chondrocytes were grown in micromass culture and were either untreated or treated with 5 ng TGF-b1/ml for 8 hours to identify genes regulated by TGF-b. Six total samples were analyzed. Three biological replicates of untreated bovine articular chondrocytes grown in micromass culture and three biological replicates of bovine articular chondrocytes grown in micromass culture and treated with 5ng TGF-b1/ ml for 8 hours.
Project description: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 RNA sequencing.
Project description:IL-1B is an important cytokine that is often found to be up-regulated during osteoarthritic and rheumatoid joint diseases. It is viewed as a catabolic factor, inducing enzymes that allow for the degradation of the cartilage extracellular matrix and also has essential roles as an autocrine and paracrine factor in fibronectin fragment-mediated degradation. It can also reduce the synthesis of the major cartilage components, type II collagen and aggrecan. On the other hand, IL-1B also has the ability to induce the growth and morphogenic factor BMP-2. During joint diseases, IL-1B is synthesized by both synovial cells and chondrocytes. Addition of IL-1 biological antagonists such as IL-1 receptor antagonists can suppress cartilage degradation in vitro. Thus, the production of IL-1B could act as the first step in mediating a cascade of other mediators in cartilage which could be relevant to the fate of the cartilage. In order to obtain a global picture of the effect of IL-1B production on human adult articular chondrocytes, we analyzed changes in gene expression induced by IL-1B by microarray analysis. We found that IL-1B has a diverse effect on gene expression profile in chondrocytes. One of the predominant responses that we observed in adult human articular chondrocytes on exposure to IL-1B is a dramatic increase in a large set of chemokines and other genes related to the inflammatory cascade. Keywords: Gene response to IL-1B (10 ng/ml) Cartilage was obtained from adult human tissue donors with above the knee amputations due to chondrosarcoma or traumatic injury or from autopsy. Chondrocytes were isolated following established protocols, maintained in high density, and treated with IL-1B (10 ng/ml). Chondrocytes treated with buffer only served as the untreated control. The experiment was carried out in duplicate. Total RNA was extracted from these chondrocytes, labeled with fluorophores (Cy3 or Cy5) and analyzed for expression changes using the Human Operon/Qiagen v3.0 oligonucleotide array. The analysis was repeated with the fluorophore dyes exchanged between the untreated and experimental RNAs.
Project description: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.
Project description:Inflammatory mediators such as interleukin 6 (IL-6) are known to activate catabolic responses in chondrocytes during osteoarthritis (OA). This study aimed to investigate the downstream targets of IL-6 in murine chondrocytes. RNA-sequencing was performed in murine articular chondrocytes treated with IL-6 in hypoxia (1%O2) or normoxia (21%O2). RNA-sequencing revealed that SerpinA3N is a major target of IL-6 in articular chondrocytes. The expression of SerpinA3N was increased in OA cartilage and further investigated.
Project description:To understand the effect of semaphorin 4D on articular chondrocytes, articular chondrocytes were stimulated with semaphorin 4D. Total RNA was analyzed by RNA-seq.
