Exploring the Crosstalk between Hydrostatic Pressure and Adipokines: An In Vitro Study on Human Osteoarthritic Chondrocytes

Abstract

Obesity is a risk factor for osteoarthritis (OA) development and progression due to an altered biomechanical stress on cartilage and an increased release of inflammatory adipokines from adipose tissue. Evidence suggests an interplay between loading and adipokines in chondrocytes metabolism modulation. We investigated the role of loading, as hydrostatic pressure (HP), in regulating visfatin-induced effects in human OA chondrocytes. Chondrocytes were stimulated with visfatin (24 h) and exposed to high continuous HP (24 MPa, 3 h) in the presence of visfatin inhibitor (FK866, 4 h pre-incubation). Apoptosis and oxidative stress were detected by cytometry, B-cell lymphoma (BCL)2, metalloproteinases (MMPs), type II collagen (Col2a1), antioxidant enzymes, miRNA, cyclin D1 expressions by real-time PCR, and β-catenin protein by western blot. HP exposure or visfatin stimulus significantly induced apoptosis, superoxide anion production, and MMP-3, -13, antioxidant enzymes, and miRNA gene expression, while reducing Col2a1 and BCL2 mRNA. Both stimuli significantly reduced β-catenin protein and increased cyclin D1 gene expression. HP exposure exacerbated visfatin-induced effects, which were counteracted by FK866 pre-treatment. Our data underline the complex interplay between loading and visfatin in controlling chondrocytes’ metabolism, contributing to explaining the role of obesity in OA etiopathogenesis, and confirming the importance of controlling body weight for disease treatment.

A Combination of Celecoxib and Glucosamine Sulfate Has Anti-Inflammatory and Chondroprotective Effects: Results from an In Vitro Study on Human Osteoarthritic Chondrocytes

“…The viability of the cells after pharmacological treatment was evaluated by MTT (3-[4,4-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide) (Sigma-Aldrich, Milan, Italy) for each experimental condition, as previously described [65]. Timing of drug treatment was selected according to the percentage of surviving cells (Figure S1).…”

Synovial Fluid Regulates the Gene Expression of a Pattern of microRNA via the NF-κB Pathway: An In Vitro Study on Human Osteoarthritic Chondrocytes

“…Apoptosis analysis was carried out considering the simultaneous staining of cells with Alexa Fluor 488 annexin-V and PI. The results were expressed as the percentage of cells positively stained by each dye (total apoptosis) [ 69 ].…”

Effects of Visfatin on Intracellular Mechanics and Catabolism in Human Primary Chondrocytes through Glycogen Synthase Kinase 3β Inactivation

“…In addition to the mechanical overloading effect, the secretion of adipokines (including chemerin, leptin, resistin, and visfatin) from white adipose tissue and synovial tissue in obese subjects has been recognized as the major sources of chronic inflammation initiating cartilage damage [ 4 , 5 , 6 , 7 ]. Among these adipokines, recent research has been focused on visfatin because of accumulating in vitro data showing its role in inducing inflammatory response (interleukin-6, tumor necrosis factor-alpha, and nuclear factor kappa B), degradative effect (oxidative stress and apoptosis), and imbalanced metabolism (MMPs upregulation and collagen II/aggrecan deregulation) in chondrocytes [ 6 , 7 , 8 ]. Moreover, clinical evidence has found a positive correlation between the serum/synovial visfatin level and OA progression in both obese and nonobese patients [ 6 , 7 , 8 , 9 , 10 , 11 ].…”

“…Among these adipokines, recent research has been focused on visfatin because of accumulating in vitro data showing its role in inducing inflammatory response (interleukin-6, tumor necrosis factor-alpha, and nuclear factor kappa B), degradative effect (oxidative stress and apoptosis), and imbalanced metabolism (MMPs upregulation and collagen II/aggrecan deregulation) in chondrocytes [ 6 , 7 , 8 ]. Moreover, clinical evidence has found a positive correlation between the serum/synovial visfatin level and OA progression in both obese and nonobese patients [ 6 , 7 , 8 , 9 , 10 , 11 ]. Visfatin is a pleiotropic protein, which is responsible for both physiological and pathophysiological processes.…”

“…Its intracellular form, called nicotinamide phosphoribosyltransferase (NAMPT), possesses enzyme activity to catalyze nicotinamide mononucleotide formation, which benefits physiological essential metabolism. Its secretory extracellular form, visfatin, usually acts as an inflammatory cytokine, which attracts much more attention because of its pathophysiological role in the development of different types of diseases, including OA [ 6 , 7 , 8 , 9 , 10 , 11 ]. The pharmacological intervention of visfatin inhibitors in different diseases, including diabetes and cancer, has been examined [ 12 ].…”