ASCs were cultured for 3 weeks in full media (serum supplemented DMEM) in the presence of 5% AH. The AH was either untreated, incubated with dextran-coated charcoal (DCC), or heat treated. (A) Both DCC (5% AH/DCC) and HT (5% AH/HT) treatments reduced ALP activity when compared to untreated AH (5% AH), with HT reducing activity to the level seen in untreated cells (Control). (B) Both DCC and HT significantly reduced the ALP induction caused by AH. The effect of DCC was partial, and was still significantly elevated compared to control cells, while HT resulted in complete loss of AH osteogenic activity. All data normalized to the response of 5% AH case. Data are mean ± SEM (n = 3 donors). * p < 0.05; ** p < 0.01; *** p < 0.001. Scale bar 50 μm.