Abstract: Mesenchymal stem cell (MSC) transplantation therapy is highly studied for the regenerative repair of cartilage defects. Low-intensity pulsed ultrasound (LIPUS) has the potential to promote chondrogenic differentiation of MSCs, which is hopeful to facilitate the transplantation therapy. However, its underlying mechanism remains unclear. Here, we investigated the promoting effects and mechanisms of LIPUS stimulation on the chondrogenic differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) and further evaluated its regenerative application value in articular cartilage defects in rats. In vitro, the suitable sound intensity of LIPUS for differentiation promotion was screened, and the key signal pathway of LIPUS-induced differentiation was clarified. In vivo, defective articular cartilage rat models were established for further validating LIPUS stimulation and hUC-MSC transplantation-based cartilage regeneration therapy. The results showed that LIPUS stimulation with specific parameters effectively promoted the expression of mature cartilage-related genes and proteins, inhibited TNF-{alpha} gene expression in hUC-MSCs, and exhibited anti-inflammation in C28/I2 cells. In addition, the articular cartilage defects of rats were significantly repaired after hUC-MSC transplantation and LIPUS stimulation. Taken together, our study demonstrated that LIPUS stimulation could realize articular cartilage regeneration due to the inhibition of TNF signaling pathway especially TNF-{alpha} gene expression down-regulation, which is of clinical value for the relief of osteoarthritis.

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