Chondrocyte Transfection and Mechanical Stimulation

Cells were grown and maintained in 6-well dishes at a starting density of 1 × 10⁵ cells/well until they became 85% confluent in DMEM supplemented with 10% FBS, before replacement with 0.5% FBS for 6hr until transfection procedure. Afterwards, chondrocytes were transfected with miR-34a, miR-146a, and miR-181a specific inhibitors (Qiagen, Hilden, Germany), at the concentration of 50 nM, or with the negative controls siRNA (NC) (Qiagen, Hilden, Germany), at the concentration of 5 nM, in serum-free medium for 24~h. Media were removed and the cells were immediately collected or exposed to cycles of low sinusoidal or static continuous HP for a period of 3~h [29 (link),30 (link)].