An in vitro approach to the chronological aging of skin by glycation of the collagen: the biological effect of glycation on the reconstructed skin model.
Glycation is a slow, nonenzymatic reaction that takes place between free amino groups in proteins primarily from lysine and a reducing sugar such as glucose or ribose. In skin, this reaction creates new residues or formations of cross-links (advanced glycation end products, AGEs) in the extracellular matrix of the dermis. The formation of these bridges between dermal molecules is supposed to be responsible for loss of elasticity or other properties of the dermis observed during aging. Glycation may therefore play an important role in chronologic aging. In order to examine this hypothesis, we have developed a reconstructed skin model made of a modified dermal compartment that is a fibroblast-contracted collagen lattice prepared with preglycated collagen. The presence of AGEs (glycoxidation products) in the skin equivalents was evidenced using specific antibodies against carboxymethyllysine (CML). Several changes were observed after collagen glycation: (1) fibroblast shape and distribution (vimentin staining) were modified; (2) extracellular matrix molecules and the dermal-epidermal junction zone seemed to be enhanced (procollagen I and III, collagen IV and VII stainings); (3) stainings for beta1 and alpha6 integrins were also increased in the epidermal cell layer; and (4) collagenase activity was increased. To verify the biological effect of glycation, we used the well-known glycation inhibitor aminoguanidine. After aminoguanidine treatment, we found a low CML amount and decreased distribution of markers previously overexpressed in glycated skin constructs. These in vitro findings were at least in part related to aging in vivo and demonstrate an actual effect of glycation in skin aging.
Pageon
Hervé
H
L'Oréal, Life Sciences, Centre Charles Zviak, 90 rue du Général Roguet, 92583 Clichy Cédex, France. hpageon@rd.loreal.com
Asselineau
Daniel
D
D000818:Animals; D001485:Basement Membrane / Q000502:physiology; D002417:Cattle; D003094:Collagen / Q000502:physiology*; D020405:Dermis / Q000502:physiology; D004817:Epidermis / Q000502:physiology; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D008954:Models, Biological; D015595:Skin Aging / Q000502:physiology*
D017127:Glycation End Products, Advanced; D003094:Collagen