Early glycation of critical fibronectin domains inhibits human dermal fibroblast migration

Failure of acute wounds to heal in diabetic patients has been found to correlate with elevated blood glucose levels at the time of surgery and correlates poorly with glycated hemoglobin, a marker for late glycation events that need weeks to amass. Few investigations have focused on the effects of short-term elevated glucose on extracellular matrix (ECM) proteins. Fibronectin (FN) is an ECM protein in the early wound provisional matrix required for fibroblast migration in this context. This study investigated the effects of early glycation of FN on dermal fibroblast migration. FN-coated plastic, FN-fibrin, and FN-hyaluronan were glycated in the presence of 3mg/ml glucose for 2 days. Cell migration in response to 100ng/ml platelet-derived growth factor-BB (PDGF-BB) was significantly impaired on glycated FN (71.8% decrease, p<0.05), FN-fibrin (40.0% decrease, p<0.05) and FN-hyaluronan (93.1% decrease, p<0.05). Although glycation had no effect on cell attachment, cell viability or apoptosis, cell spreading was markedly inhibited. In addition, MALDI-TOF was performed and showed shift in mass peak indicating the presence of glucose molecules crosslinked to FN-heparin binding domain. These data strongly suggests that glycation of FN, as little as 2 days, inhibits cell traction resulting in loss of motogenic potential of these cells.