Fig. 3

Alcian blue staining of osteochondral explants. Incised osteochondral explants (n = 6 explants per group per time point) were grown in DMEM-F12 medium supplemented with 10% FBS, 1× antibiotic-antimycotic solution and 50 μg/ml L-ascorbic acid in the presence or absence of IL-6 or TNFα under cLIUS at 14 kPa (5 MHz, 2.5 Vpp), 20 min, 4 times/day. Non-cLIUS-stimulated explants (n = 6 explants per group per time point) served as controls for each treatment condition. Explants were fixed in 10% neutral buffered formalin and embedded in paraffin. The panel shows 4 μm sections of osteochondral explants at the interfacial region stained with alcian blue (pH 1) after 14 and 28 days in culture at 20× magnification. Scale bar represents 400 μm. Insets depict the whole section imaged at 2× magnification. Black arrows in the insets depict the sites of incision