We investigated whether focused low-intensity pulsed ultrasound (FLIPUS) affects extracellular matrix (ECM) production in osteoarthritic (OA) rabbits by decreasing chondrocyte apoptosis and pro-inflammatory mediators. An OA model using New Zealand White rabbits (N = 30) and 30 normal rabbits were randomized into three groups (2-, 4- and 8-wk groups; n = 10 knees each). A knee from each rabbit was randomly selected to receive FLIPUS and the other knee received a sham treatment as a control. Another 30 normal rabbits were blank controls. We measured ECM degradation, joint effusion volume and levels of prostaglandin E2 and nitric oxide. Also, ratios of chondrocyte proliferation and apoptosis were calculated. Compared with sham stimulation, FLIPUS attenuated release of type II collagen and proteoglycans and reduced chondrocyte apoptosis as well as total joint effusion volume and significantly alleviated OA-induced accretion of prostaglandin E2 and nitric oxide in the synovial fluid. FLIPUS application promoted ECM production in OA through down regulation inflammatory mediators, joint effusion volume and chondrocyte apoptosis.