Expression of Dermal Extracellular Matrix Proteins in a Newly Developed Full-Thickness Skin Model

摘要

In order to analyze the metabolism of the extracellular matrix in vitro, we have developed a multilayered full-thickness skin equivalent. Using immunohisto-logical and PCR techniques we investigated the expression of elastin, fibrillin-1 and fibulin-5 (DANCE), the main components of the elastic system, as well as of MMPs in the dermal tissue of the skin models.Elastin was detected for the first time 7 days after cultivation at the air-liquid interface (ALI), which exactly correlates with the beginning of elastin mRNA expression. With proceeding culture time, elastin-positive fibres accumulated in the dermal compartment with shorter fibres preferably directly beneath the dermo-epidermal junction (DEJ) and longer fibres in the deeper dermal regions. Fibrillin-1 as well as fibulin-5 could be co-localized with elastin on distinct fibres of the dermal elastic network. MMP-2 was detected predominantly intracellularly in the fibroblasts by a specific antibody.These results clearly show that fundamental elements of the dermal elastic network are correctly synthesized and assembled in the skin model, thus corroborating the high level of correspondence with native human skin.