Abstract
Osteoarthritis (OA), one of the most common joint disease, affects more than 80% of the population aged 70 or over. Mesenchymal stem cells (MSCs) show multi-potent differentiation and self-renewal capability, and, after exposure to an inflammatory environment, also exhibit immunosuppressive properties. In this study, we have used a model of lipopolysaccharide (LPS)-stimulated chondrocytes to evaluate MSC anti-inflammatory efficacy. The anti-inflammatory mechanism was tested in two cell-contained culture systems: (i) MSC-chondrocyte indirect contact system and (ii) MSC-chondrocyte direct contact system, and one cytokine-only culture system: MSC-conditioned medium (CM) system. Results showed that MSCs reduced chondrocyte inflammation through both paracrine secretion and cell-to-cell contact. The inflammation-associated, and free-radical-related genes were down-regulated significantly in the direct contact system on 24 h, however, the TNF-α. IL-6 were upregulated and aggrecan, COLII were downregulated on 72 h in direct contact system. Moreover, we found CM produced by MSC possess well therapeutic effect on inflammatory chondorcyte, and the 10-fold concentrated MSC-conditioned medium could down-regulated chondorcyte synthesis inflammation-associated, and free-radical-related genes, such as TNF-α, IL-1β, IL-6 and iNOS even treated for 72 h. In conclusion, MSC-CM showed great potential for MSC-based therapy for OA.
Highlights
MethodsMaterialsThe Animal Research was approved by Far Eastern Memorial Hospital IACUC, and the approval number was 102-02-06-A
Osteoarthritis (OA), one of the most common degenerative joint diseases, affects more than 80% of the population aged 70 or over
Full thickness porcine articular cartilage was minced, washed three times in sterile Phosphate-buffered saline (PBS) containing 5% (100x) antibiotic-antimycotic solution and twice in PBS at room temperature, chondrocytes were isolated by digestion for 18 h with 0.5% collagenase in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% (100x) antibiotic-antimycotic solution at 37 ̊C in a 5% CO2 incubator
Summary
MaterialsThe Animal Research was approved by Far Eastern Memorial Hospital IACUC, and the approval number was 102-02-06-A. Fresh porcine knees were purchased from a traditional market and kept skin intact for 2–3 h on ice until harvesting cartilage under aseptic conditions. Full thickness porcine articular cartilage was minced, washed three times in sterile PBS containing 5% (100x) antibiotic-antimycotic solution and twice in PBS at room temperature, chondrocytes were isolated by digestion for 18 h with 0.5% collagenase in DMEM supplemented with 10% FBS and 1% (100x) antibiotic-antimycotic solution at 37 ̊C in a 5% CO2 incubator. The porcine chondrocytes were resuspended, and washed twice, in PBS, were cultured in DMEM supplemented with 10% FBS in 100 mm diameter culture dishes at a density of 5 x 105 cells/ dish.
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