title: Protective effect of matrix metalloproteinase inhibitors against epidermal basement membrane damage: skin equivalents partially mimic photoageing process.
abstract: BACKGROUND
  The epidermal basement membrane (BM) plays important roles in adhesion between epidermis and dermis, and in controlling epidermal differentiation. The BM has been reported to be damaged in sun-exposed skin. Although matrix metalloproteinases (MMPs) are believed to be involved in the BM damage, there is no good in vitro model for examining BM damage by MMPs or for exploring methods to protect the BM.
  
  
  OBJECTIVE
  To examine the involvement of MMPs in BM damage and approaches to protect the BM from such damage by using an in vitro skin-equivalent (SE) model.
  
  
  METHODS
  SE was prepared by culturing human keratinocytes on contracted collagen gel including human fibroblasts. MMP-1, -2, -3 and -9, laminin 5 and type IV and VII collagens were determined by specific sandwich ELISAs, and MMP-2 and MMP-9 were analysed by gelatin zymography. Histological examination of SE was also carried out.
  
  
  RESULTS
  Despite production of BM components such as laminin 5 and type IV and VII collagens in SEs, BM was rarely observed at the dermal-epidermal junction. Several MMPs, such as MMP-1, -2, -3 and -9, were observed to be present in conditioned media and some of them were in active forms. Tissue inhibitor of metalloproteinase (TIMP)-2 was not detected, although TIMP-1 was present. Synthetic MMP inhibitors, CGS27023A and MMP-inhibitor I, which inhibit MMP-1, -2, -3 and -9, markedly augmented deposition of laminin 5 and type IV and VII collagens at the dermal-epidermal junction, resulting in the formation of continuous epidermal BM.
  
  
  CONCLUSIONS
  Our results indicate that MMPs are involved in the degradation of BM in SEs, and that MMP inhibitors exert a protective effect against BM damage.
authors.affiliation: Skin Biology Research Laboratories, Shiseido Life Science Research Centre, Yokohama, Japan 236-8643. satoshi.amano@to.shiseido.co.jp
authors.forename: S
authors.initials: S
authors.lastname: Amano
authors.forename: Y
authors.initials: Y
authors.lastname: Ogura
authors.forename: N
authors.initials: N
authors.lastname: Akutsu
authors.forename: Y
authors.initials: Y
authors.lastname: Matsunaga
authors.forename: K
authors.initials: K
authors.lastname: Kadoya
authors.forename: E
authors.initials: E
authors.lastname: Adachi
authors.forename: T
authors.initials: T
authors.lastname: Nishiyama
mesh_terms: D001485:Basement Membrane / Q000187:drug effects / Q000201:enzymology / Q000528:radiation effects*; D015815:Cell Adhesion Molecules / Q000032:analysis; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000032:analysis; D024143:Collagen Type VII / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004817:Epidermis / Q000201:enzymology / Q000528:radiation effects / Q000648:ultrastructure*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000032:analysis; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D008854:Microscopy, Electron; D008954:Models, Biological; D009842:Oligopeptides / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D011719:Pyrazines / Q000494:pharmacology*; D015595:Skin Aging; D013449:Sulfonamides / Q000494:pharmacology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis; D014466:Ultraviolet Rays / Q000009:adverse effects*; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D024141:Collagen Type IV; D024143:Collagen Type VII; D006877:Hydroxamic Acids; D020781:Matrix Metalloproteinase 1; D020778:Matrix Metalloproteinase 2; D019278:Matrix Metalloproteinase 3; D020780:Matrix Metalloproteinase 9; D009842:Oligopeptides; D011480:Protease Inhibitors; D011719:Pyrazines; D013449:Sulfonamides; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D000097794:Kalinin; C095170:4-aminobenzoyl-glycyl-prolyl-leucyl-alanine hydroxamic acid; C097658:CGS 27023A