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Decellularized allogeneic heart valves have been successfully used for transplantation in patients 3. In cardiovascular surgery, decellularized heart valves have been used to replace malfunctioning heart valves 2. Taken together, we conclude that sucrose can minimize oxidative damage caused by freeze-drying, and that subsequent dried storage has little effects on the overall biochemical composition of heart valve scaffolds.ĭecellularized tissues have been successfully used in a variety of tissue engineering applications 1. However, fresh control, freeze-dried and stored samples could not be distinguished from one another, which implies that these groups are very similar in terms of their biomolecular fingerprints. A feed forward artificial neural network model could accurately classify H 2O 2 and FeCl 3 treated samples. FTIR analysis shows that fresh control, freeze-dried and stored heart valve specimens cannot be distinguished from one another, whereas H 2O 2- and FeCl 3-treated samples could be distinguished in some tissue section. Dried specimens that were stored at 4 ☌ depict little to no oxidative damage during storage for up to 2 months. NBT staining revealed that sucrose at a concentration of at least 40% (w/v) is needed to prevent oxidative damage during freeze-drying. In this study, decellularized porcine aortic heart valves were analyzed by nitroblue tetrazolium (NBT) staining and Fourier transform infrared spectroscopy (FTIR) to identify oxidative damage during freeze-drying and subsequent storage as well as after treatment with H 2O 2 and FeCl 3. Freeze-drying can be used to ensure off-the-shelf availability of decellularized heart valves for cardiovascular surgery.