Modulation of the Effect of Transforming Growth Factor-β3 by Low-Intensity Pulsed Ultrasound on Scaffold-Free Dedifferentiated Articular Bovine Chondrocyte Tissues.

The aim of this study was to evaluate how low-intensity pulsed ultrasound (LIPUS) modulates the effect of transforming growth factor-β3 (TGF-β3) on the differentiation of scaffold-free dedifferentiated bovine articular chondrocyte tissues toward a cartilage-like phenotype. Specifically, the effect of these stimuli on the expression of hypertrophic markers collagen type I, collagen type X, and cartilage-degrading collagenase gene expression for a scaffold-free model was analyzed. A bioreactor that applied LIPUS directly from the transducer through a silicone gel to a six-well plate containing the tissues allowed simple, sterile, and large-scale experiments. Tissues were subjected to LIPUS of 55 mW/cm(2) in a 200 μs burst sine wave of 1 MHz over a 10-day period with or without TGF-β3 (10 ng/mL). Tissues exposed to TGF-β3 had significantly increased glycosaminoglycan and total collagen protein production along with upregulated cartilage-specific gene expression, resulting in tissues with a higher Young's Modulus. However, these tissues had also upregulated gene expression for hypertrophic markers collagen type I, collagen type X, MMP-1, MMP-13, MMP-2, and also an increase in the phosphorylation of p38. The expression of these matrix-degrading enzymes was remediated by hypertrophic development and differentiate dedifferentiated bovine articular chondrocytes towards a chondrogenic lineage allowing it to be a valuable tool in cartilage tissue engineering.