Aortic valve stenosis (AVS) is one of the most common valve diseases, characterised by calcification. Currently the treatment only option is surgical intervention. Inflammation plays a key role in initiating calcification of valve interstitial cells (VICs). Proinflammatory mediators activate VICs leading to osteogenic differentiation. Recent studies have indicated that AVS has a higher prevalence in diabetes patients. Research indicates that cardiovascular disease in diabetes patients is increased due to high glucose and the formation of advanced glycation end products (AGEs) in blood. The current study aims to understand the link between high glucose and AGEs on VICs in an osteogenic environment. Porcine VICs were isolated from the pig heart obtained from the abattoir and cultured under sterile conditions. Alizarin red stain was used to determine calcium deposition as an indicator of osteogenic differentiation. After 7 days in osteogenic medium, calcification was clearly visible in VICs and this timescale was subsequently used for the study. Inducible cyclooxygenase 2 (COX2) is a key mediator of pro-inflammatory pathways and may play a role in the inflammation of VICs. Immunoblotting was used to determine COX-2 protein expression. COX-2 expression was not changed in VICs incubated in osteogenic medium for 7 days. However AGEs (100ug/ml) significantly increased COX2 expression in osteogenic medium (7.50±3.11 fold increase vs control, P<0.05, n=6). Interestingly, AGEs incubation for 7 days had no effect of COX-2 expression in non-osteogenic medium. The effects of high glucose were examined. In non-osteogenic medium, high glucose (4500mg/ml) significantly increased the expression of COX2 (4.06±0.93 fold increase vs normal 1000mg/L glucose control after 7 days, P<0.05, n=5). Furthermore, the combination of high glucose and AGEs for 7 days in non-osteogenic medium had a synergistic effect, significantly inducing COX2 expression compared to high glucose alone or AGEs alone (4.51±1.13 fold increase vs AGE incubation; 2.54±0.46 fold increase vs high glucose incubation, P<0.05, n=4). Finally, all 3 factors were examined together (AGEs, high glucose and osteogenic medium). The combination of high glucose and AGEs in osteogenic medium significantly increased COX2 expression compared to all other factors alone (for example, 4.90±1.49 fold increase compared to AGEs alone, P<0.05, n=6). In conclusion, this data reveals that although calcification in vitro does not change expression of the pro-inflammatory enzyme COX-2 in VICs, the addition of either AGEs or high glucose under these conditions induces COX2 expression. In combination, these factors have a significant synergistic effect. Thus, diabetes patient with increased may have an increased inflammation in aortic valves, leading to an enhanced progression of AVS.