Glycation by glyoxal leads to profound changes in the behavior of dermal fibroblasts

Abstract

IntroductionDiabetes is a worldwide health problem that is associated with severe complications. Advanced Glycation End products (AGEs) such as Nε-(carboxymethyl)lysine, which result from chronic hyperglycemia, accumulate in the skin of patients with diabetes. The effect of AGEs on fibroblast functionality and their impact on wound healing are still poorly understood.Research design and methodsTo investigate this, we treated cultured human fibroblasts with 0.6 mM glyoxal to induce acute glycation. The behavior of fibroblasts was analyzed by time-lapse monolayer wounding healing assay, seahorse technology and atomic force microscopy. Production of extracellular matrix was studied by transmission electronic microscopy and western blot. Lipid metabolism was investigated by staining of lipid droplets (LDs) with BODIPY 493/503.ResultsWe found that the proliferative and migratory capacities of the cells were greatly reduced by glycation, which could be explained by an increase in fibroblast tensile strength. Measurement of the cellular energy balance did not indicate that there was a change in the rate of oxygen consumption of the fibroblasts. Assessment of collagen I revealed that glyoxal did not influence type I collagen secretion although it did disrupt collagen I maturation and it prevented its deposition in the extracellular matrix. We noted a pronounced increase in the number of LDs after glyoxal treatment. AMPK phosphorylation was reduced by glyoxal treatment but it was not responsible for the accumulation of LDs.ConclusionGlyoxal promotes a change in fibroblast behavior in favor of lipogenic activity that could be involved in delaying wound healing.

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

“…AGE formation induces protein modifications and oxidative stress by binding with their target receptor, RAGE. As a result, many mechanisms involved in wound healing such as cell proliferation, migration and differentiation, neurogenic inflammation, collagen deposition, and microvascular morphology are adversely affected (Guillon et al, 2021; Sugimoto et al, 2008; Sveen et al, 2013).…”

Effect of a Topical Collagen Tripeptide on Antiaging and Inhibition of Glycation of the Skin: A Pilot Study

“…Glycation and production of AGEs further debilitate the function of collagen, making it stiffer and more brittle [17]. The glycation process also leads to profound changes in the behavior of dermal fibroblasts, reducing their proliferation and migration, while simultaneously disrupting collagen I maturation and preventing collagen deposition in the extracellular matrix [18].…”

Cutaneous changes in diabetic patients: Primed for aberrant healing?

“…111 In vitro treatment of human fibroblasts with glyoxal, a glycation reaction product, revealed a decrease in fibroblast proliferation, increased tensile strength, increased adherence to ECM, downregulated focal adhesion kinase and exhibited failure to extend filipodia, demonstrating multiple points of hindrance to fibroblast migration and proliferation as a result of AGE treatment. 112 Treatment of fibroblasts with rosiglitazone, a peroxisome proliferator-activated receptor-gamma ligand, attenuated the effects of AGE in vitro. 113 Connective tissue growth factor (CTGF), a key player in fibroblast-ECM production and angiogenic factor, has been identified as a potential mediator between increased AGE and tissue fibrosis in diabetes.…”