Fig. 12.

Aminoguanidine (AG) protects against glyoxal-induced inhibition of in vitro angiogenesis in endothelial cells. BPAECs were grown up to 80% confluence in complete medium in two separate flasks. Both flasks of cells were starved in minimal media (MEM) at 37°C for 30 min and following that AG HCl (5 mM) in MEM was added to one of the flask and continued to incubate for an additional 45 min. The cells from both flasks were then added to the Matrigel-**coated plate [Rich media + Glyoxal (1 mM)] as described in the Materials and methods section. a BPAECs that were pre-treated with AG HCl were added to either 1 ml of minimal media (Minimal media), 1 ml of rich media (Rich media), or to 1 ml of rich media supplemented with Glyoxal (1 mM) [Rich media + AG (5 mM) + Glyoxal (1 mM)]. The cells were allowed to incubate for 12 h at 37°C and subsequently analyzed for the development of tubes connecting one colony to the next. The media was removed and the cells were fixed with 4% paraformaldehyde. b Four digital images were captured per well at the same locations in each well and the tubes were counted in a blinded fashion. The data represent n = 6 for each condition. The statistical analysis was performed using an ANOVA with SYSTAT12 software (San Jose, CA)