EFFECTS OF HYPERGLYCEMIA ON WOUND HEALING IN THREE-DIMENSIONAL CELL CULTURE

Hyperglycemia is the most deteriorating factor causing pathological and functional changes in patients with Diabetes mellitus including non-healing wounds, damaging feet and legs. Despite the intensive studies, the molecular mechanisms of action of high glucose concentrations on cells are poorly defined. Here, we used an in vitro model system, based on experimentally wounded three-dimensional fibroblast cell culture, in order to study wound healing under normoglycemic and hyperglycemic conditions. Following the wound closure for thirteen days by phase-contrast microscopy demonstrated that high glucose delayed the healing process. This was due to decreased fibroblast trans-differentiation into myofibroblasts as judged by reduced expression of the myofibroblasts marker alpha-smooth muscle actin. Probing the activity of TGF-beta signalling pathway, which is responsible for fibroblast activation, showed deficiency in Smad2/3 phosphorylation even after addition of exogenous TGF-beta 1. These results indicate that high glucose concentrations are able to attenuate TGF-beta signal transduction pathway and this effect should be considered an important contributing factor when therapies for diabetic wound healing are designed.