Reconstruction of basement membrane in skin equivalent; role of laminin-1

• Published in: Archives of Dermatological Research Vol. 293; no. 7; pp. 356 - 362
• Main Authors: JAE YOUN YI, YONG HA YOON, YOUNG SOOK SON, HYUN SOOK PARK, CHUN HO KIM, CHANG HWAN KIM, HYUN JOO KANG, LEE, Eunah, YOUN YOUNG KIM, YONG JAE JIN, TAE HWAN KIM
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.07.2001; Springer Nature B.V
• Subjects: 3T3 Cells; Animals; Antigens, Surface - physiology; Basal cells; Basement Membrane - physiology; Basement membranes; Biological and medical sciences; Cell surface; Collagen; Collagen (type IV); Collagen Type IV - physiology; Dermatology; Extracellular matrix; Fibronectin; Fibronectins - pharmacology; Gelatinase A; Gelatinase B; Integrin alpha6beta4; Integrins - physiology; Keratinocytes; Keratinocytes - drug effects; Keratinocytes - physiology; Laminin; Laminin - antagonists & inhibitors; Laminin - physiology; Matrix Metalloproteinase 2 - physiology; Matrix Metalloproteinase 9 - physiology; Medical sciences; Membrane structure; Mice; Skin; Skin involvement in other diseases. Miscellaneous. General aspects; Skin, Artificial; Swine; Wound healing; Human; Reconstruction; Cell culture; Laminin; Collagen; Migration; Equivalent; Keratinocyte; Skin; Basement membrane
• ISSN: 0340-3696; 1432-069X
• DOI: 10.1007/s004030100234

To reconstruct the basement membrane in a skin equivalent, the epidermodermal interface was coated with porcine type IV collagen and mouse laminin-1 at various ratios before keratinocyte seeding. Laminin-1, a component of the basement membrane, induced massive infiltration of keratinocytes into the dermal equivalent, while type IV collagen induced discrete demarcation between dermal and epidermal compartments without any infiltrating cells. Immunohistochemical staining indicated that the laminin-induced infiltrating cells expressed endogenous type IV collagens at the cell periphery, which were not incorporated into the basement membrane structure. The infiltrating cells did not express fibronectin receptor alpha5beta1 integrin but showed MMP-9 secretion and cell surface associated MMP-2. However, when laminin-1 was preincubated with type IV collagen, laminin-1-induced keratinocyte infiltration as well as MMP-9 induction were almost completely suppressed to basal levels. Therefore, replenishment of the type IV collagen lattice seemed to cause laminin-stimulated cells to anchor to the lattice, in a similar manner to the basal cells on the basement membrane of normal skin. Our study suggests that the molar ratio of basement membrane components may determine the behavior of basal cells within the wound healing microenvironment, which is probably regulated either by extracellular matrix deposition or degradation.