During cutaneous wound healing, re-epithelialization undergoes by the migration and proliferation of keratinocytes(KC) at the wound site. Signals for the stimulation of KC include biologically active substances such as growth factors, chemotactic
factors and changes in molecular structure of extracellular matrix, Migration and proliferation of KC are stimulated by fibronectin and soluble growth factors such as hepatocyte growth factor(HGF) and epidermal growth factor(EGF).
This study was performed to investigate the effect of HGF on keratinocyte migration and whether fibronection secretion correlate to stimulation of the KC migration by HGF and/or EGF. Primary kertinocytes culture from neonatal foreskins were done
using
the complete M154 medium. We performed the quantitative analysis of KC migration by cell count with Giemsa stain and the fibronectin secretion of media by sandwich-ELISA method after half-removal of cultured monolayer KC in 6 well plate.
@ES The results were as follows:
@EN 1. The number of migrated KC were 57.7±11.05 with HGF(2.5ng/ml), 55.5±7.06 with EGF(10ng/ml), 234.7±19.79 with HGF(2.5ng/ml) and EGF(10ng/ml) while 30.1±9.37 without HGF and EGF, respectively after 24 hours. HGF and/or EGF significantly
increased KC migration compared with the control(P<0.05).
2. The fibronectin secretion of KC in KBM(keratinocyte basal media) were observed 10.0±0.82 ng/ml/3. 8×10E4 cell with HGF(2.5ng/ml), 10.8±0.5 ng/ml/5.3×10E4 cell with EGF(10ng/ml), 21.5±0.58 ng/ml/6.2×10E4 cell with HGF and EGF. Compare
with
the
control, HGF and/or EGF significantly increased fibronectin secretion after 24 hour(P<0.05).
Therefore, HGF and/or EGF increased both the migration and fibronectin secretion of keratinocyte significantly compared with control group(P<0.05). In addition, synergistic effect of HGF with EGF was also observed in cultured human keratinocytes.
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