In vitro glycation of an endothelialized and innervated tissue-engineered skin to screen anti-AGE molecules

• Published in: Biomaterials 2015-05, Vol.51, p.216-225
• Main Authors: Cadau, Sébastien, Leoty-Okombi, Sabrina, Pain, Sabine, Bechetoille, Nicolas, André-Frei, Valérie, Berthod, François
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Advanced glycation end products; Age; Angiogenesis; Animals; Capillaries - drug effects; Capillarity; Carboxymethyl lysine; Cattle; Cell Differentiation - drug effects; Cells, Cultured; Collagen - pharmacology; Cytoskeleton - drug effects; Cytoskeleton - metabolism; Dentistry; Dermis - blood supply; Dermis - drug effects; Endothelial Cells - cytology; Endothelial Cells - drug effects; Fibroblasts - cytology; Fibroblasts - drug effects; Formations; Glycation End Products, Advanced - antagonists & inhibitors; Glycosylation - drug effects; Glyoxal; Glyoxal - pharmacology; Guanidines - pharmacology; Humans; Keratinocytes - cytology; Keratinocytes - drug effects; Lysine - analogs & derivatives; Lysine - metabolism; Mice; Nerves; Networks; Neurons - cytology; Neurons - drug effects; Protective Agents - pharmacology; Screens; Skin; Skin - drug effects; Skin - innervation; Surgical implants; Thiazoles - pharmacology; Tissue Engineering - methods
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2015.01.066

Abstract Glycation is one of the major processes responsible for skin aging through induction of the detrimental formation of advanced glycation end-products (AGEs). We developed an innovative tissue-engineered skin combining both a capillary-like and a nerve networks and designed a protocol to induce continuous AGEs formation by a treatment with glyoxal. We determined the optimal concentration of glyoxal to induce AGEs formation identified by carboxymethyl-lysin expression while keeping their toxic effects low. We showed that our tissue-engineered skin cultured for 44 days and treated with 200 μ m glyoxal for 31 days displayed high carboxymethyl-lysine expression, which induced a progressively increased alteration of its capillary and nerve networks between 28 and 44 days. Moreover, it produced an epidermal differentiation defect evidenced by the lack of loricrin and filaggrin expression in the epidermis. These effects were almost completely prevented by addition of aminoguanidine 1.5 m m , an anti-glycation compound, and only slightly decreased by alagebrium 500 μ m , an AGE-breaker molecule. This tissue-engineered skin model is the first one to combine a capillary and nerve network and to enable a continuous glycation over a long-term culture period. It is a unique tool to investigate the effects of glycation on skin and to screen new molecules that could prevent AGEs formation.