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Impact of Hydration on MMP‐Activity
Author(s) -
Tandara Andrea A.,
Mustoe Thomas A.,
Mogford Jon E.
Publication year - 2004
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/j.1067-1927.2004.0abstractxp.x
Subject(s) - hek 293 cells , matrix metalloproteinase , chemistry , secretion , in vitro , paracrine signalling , cell culture , blot , microbiology and biotechnology , biochemistry , biology , receptor , gene , genetics
Hydration of keratinocytes modifies the levels of cytokines they secrete, which in turn impacts the secretory behaviour of dermal fibroblasts. In an in vitro coculture model, conditioned media (CM) collagen content was decreased 44% when keratinocytes were hydrated. We hypothesized that this is partly due to increased MMP‐activity. We used the same coculture model to study changes in MMP‐activity and TIMP secreted by keratinocytes as well as by fibroblasts in monoculture and in coculture in relation to air‐treatment or hydration of keratinocytes. Stratified human epidermal keratinocytes (HEK) and confluent human dermal fibroblasts (HDF) were cocultured for 72 h under serum‐free conditions. HEK were either kept at the air‐interface or hydrated. CM was assayed for MMP‐1, −2, −9, TIMP‐1 and −2 were assayed using zymograms, western blotting, and ELISA. MMP‐1, secreted by both cell types, increased significantly in cocultures compared to monocultures (4‐fold in the air‐treated group, 26‐fold in the hydrated group). MMP‐2, secreted mainly by HDFs, was significantly increased by coculture (hydration: 2.4‐fold, air: 2.8‐fold). MMP‐9, predominantly secreted by air‐treated HEKs and was significantly decreased in hydrated monoculture (76%) and coculture. HEK‐monoculture hydration also significant decreased MMP‐1 (86%) and MMP‐2 (81%) activity. HDF‐secreted TIMP‐1 expression was significantly increased by coculture and was unaffected by hydration. Our findings demonstrate that paracrine interactions between HEK and HDF modify MMP activity and that HEK hydration significantly effects on MMP activity. The findings provide insight into the role of hydration on HEK and HDF ctivity during the wound healing process.