Glycerol preservation is an effective method for long-term preservation of skin allografts and has a potential use in preserving arterial allografts. We evaluated the effect of glycerol concentration and incubation period on vessel-wall integrity of rat aortic allografts. No significant differences were measured in breaking strength (2.3 +/- 0.3 N) and bursting pressure (223 +/- 32 kPa) between standard glycerolized and control segments (1.7 +/- 0.3 N, 226 +/- 17 kPa). Isometric tension measurements showed complete lack of functional contraction and relaxation capacity in allograft segments prepared according to all preservation protocols. Morphologically, thickness of the vessel-wall media diminished after preservation using low (30/50/75%) or high (70/85/98%) concentrations of glycerol, as compared to control segments (i.e. 81 +/- 2.4 microm, 95 +/- 5.6 microm and 125 +/- 3.5 microm, respectively). Confocal microscopy and Fourier analysis demonstrated that vascular collagen and elastin bundle orientation had remained unaltered. Electron microscopy showed defragmentation of luminal endothelial cells. In conclusion, glycerol preservation of rat aorta resulted in an acellular tissue matrix, which maintained biomechanical integrity and extracellular matrix characteristics. The next step in the investigation will be to test the concept of glycerol preservation of arterial allografts in a vascular transplantation model.