Diazoxide ameliorates severity of experimental osteoarthritis by activating autophagy via modulation of the osteoarthritis‐related biomarkers

Abstract

Accumulating evidence suggests that autophagy plays a protective role in chondrocytes and prevents cartilage degeneration in osteoarthritis (OA). The objective of this study was to investigate the effect of diazoxide on chondrocyte death and cartilage degeneration and to determine whether these effects are correlated to autophagy in experimental OA. In this study, a cellular OA model was established by stimulating SW1353 cells with interleukin 1β. A rat OA model was generated by transecting the anterior cruciate ligament combined with the resection of the medial menisci, followed by treatment with diazoxide or diazoxide combination with 3-methyladenine. The percentage of viable cells was evaluated using calcein-acetoxymethyl/propidium iodide double staining. The messenger RNA expression levels of collagen type II alpha 1 chain (COL2A1), matrix metalloproteinase 13 (MMP-13), TIMP metallopeptidase inhibitor 1 (TIMP-1), and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) were determined using quantitative real-time polymerase chain reaction. The cartilage thickness and joint space were evaluated using ultrasound. SW1353 cell degeneration and autophagosomes were observed using transmission electron microscopy. The expression levels of microtubule-associated protein 1 light chain 3 (LC3), beclin-1, P62, COL2A1, and MMP-13 were evaluated using immunofluorescence staining and Western blot analysis. Diazoxide significantly attenuated articular cartilage degeneration and SW1353 cell death in experimental OA. The restoration of autophagy was observed in the diazoxide-treated group. The beneficial effects of diazoxide were markedly blocked by 3-methyladenine. Diazoxide treatment also modulated the expression levels of OA-related biomarkers. These results demonstrated that diazoxide exerted a chondroprotective effect and attenuated cartilage degeneration by restoring autophagy via modulation of OA-related biomarkers in experimental OA. Diazoxide treatment might be a promising therapeutic approach to prevent the development of OA.

Conditioned medium of mesenchymal stem cells delays osteoarthritis progression in a rat model by protecting subchondral bone, maintaining matrix homeostasis, and enhancing autophagy

“…In , Carames et al discovered autophagy as a protective mechanism in normal cartilage but was hampered by OA, resulting in cartilage degradation. Since then, the importance of autophagy in OA therapy has been proved repeatedly (Meng et al, ). Furthermore, Tsai et al reported that MSCs–CM ameliorated spinal cord injury by promoting autophagy.…”

Alcohol‐triggered silk fibroin hydrogels having random coil and β‐turn structures enhanced for cytocompatible cell response

“…At each day till 7 days, the number of cells was evaluated by MTT assay . In this case, cellular viability of the hydrogels was confirmed microscopically by live/dead assay . The control groups of this study were similarly compared using a tissue culture plate (TCP) and the non‐alcoholic silk fibroin hydrogel (No alc).…”

Autophagy in the pathogenesis and therapeutic potential of post-traumatic osteoarthritis

“…found that hydroxytyrosol can inhibit the inflammatory response of chondrocytes through SIRT6-mediated autophagy [ 138 ]. Diazoxide, as one of the potential drugs to prevent OA, can significantly improve the severity of experimental PTOA, which was related to the restoration of impaired autophagy [ 139 ]. An increasing number of studies has found that different compounds can cause changes in autophagy levels.…”