In vitro glycation of a tissue‐engineered wound healing model to mimic diabetic ulcers

Abstract Diabetic foot ulcers are a major complication of diabetes that occurs following minor trauma. Diabetes‐induced hyperglycemia is a leading factor inducing ulcer formation and manifests notably through the accumulation of advanced glycation end‐products (AGEs) such as N‐carboxymethyl‐lysin. AGEs have a negative impact on angiogenesis, innervation, and reepithelialization causing minor wounds to evolve into chronic ulcers which increases the risks of lower limb amputation. However, the impact of AGEs on wound healing is difficult to model (both in vitro on cells, and in vivo in animals) because it involves a long‐term toxic effect. We have developed a tissue‐engineered wound healing model made of human keratinocytes, fibroblasts, and endothelial cells cultured in a collagen sponge biomaterial. To mimic the deleterious effects induced by glycation on skin wound healing, the model was treated with 300 µM of glyoxal for 15 days to promote AGEs formation. Glyoxal treatment induced carboxymethyl‐lysin accumulation and delayed wound closure in the skin mimicking diabetic ulcers. Moreover, this effect was reversed by the addition of aminoguanidine, an inhibitor of AGEs formation. This in vitro diabetic wound healing model could be a great tool for the screening of new molecules to improve the treatment of diabetic ulcers by preventing glycation.