Expression of Hepatocyte Growth Factor-Like Protein in Human Wound Tissue and Its Biological Functionality in Human Keratinocytes.

James Glasbey,David Bosanquet,Andrew Sanders,Fiona Ruge,Wen Jiang,Keith Harding

doi:10.3390/biomedicines3010110

James Glasbey, David Bosanquet + Show 4 more

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https://doi.org/10.3390/biomedicines3010110

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Journal: Biomedicines	Publication Date: Feb 4, 2015
Citations: 41	License type: CC BY 4.0

Affiliation: Capital Medical University, Cardiff University

Abstract

Hepatocyte growth factor-like protein (HGFl) and its receptor, Recepteur d'Origine Nantais (RON), have been implicated in the development of wound chronicity. HGFl and RON expression was detected in acute wound tissue, chronic wound tissue and in normal skin using quantitative polymerase chain reaction (Q-PCR). HGFl and RON expression was also assessed in chronic healing and chronic non-healing wound tissues using Q-PCR and immunohistochemical staining. Expression was similarly detected in the HaCaT immortalized human keratinocyte cell line using reverse transcription polymerase chain reaction (RT-PCR). rhHGFl was used to assess the impact of this molecule on HaCaT cell functionality using in vitro growth assays and electric cell-substrate impendence sensing (ECIS) migration assays. HGFl and RON transcript expression were significantly increased in acute wound tissue compared to chronic wound tissue and were also elevated, though non-significantly, in comparison to normal skin. Minimal expression was seen in both healing and non-healing chronic wounds. Treatment of HaCaT cells with rhHGFl had no effect on growth rates but did enhance cell migration. This effect was abolished by the addition of a phospholipase C gamma (PLCγ) small molecule inhibitor. The increased expression of HGFl and RON in acute, healing wounds and the pro-migratory effect of HGFl in an in vitro human keratinocyte model, may indicate a role for HGFl in active wound healing.

Highlights

Cutaneous wound healing is a complex and multifactorial process
Hepatocyte growth factor-like protein (HGFl) expression was found to be significantly higher in acute wound tissue (Median 0.058, IQR 0.013–4225) compared to chronic wound tissues (Median 0.0004, IQR 0.0002–0.0068, p < 0.05; Figure 1A)
A similar pattern was observed in Recepteur d'Origine Nantais (RON) transcript expression (Figure 1B) where higher levels of transcript were seen in acute wound tissues than in chronic wound tissues and normal skin

Summary

Introduction

Cutaneous wound healing is a complex and multifactorial process. Numerous cellular and molecular players contribute to the ordered process of wound healing [1]. A number of local and systemic aberrations of these healing pathways have been implicated in wound chronicity [2]. Chronic non-healing wounds pose a significant problem to European healthcare providers, with an estimated 3.55–3.70 patients per 1000 population currently undergoing wound treatment [3]. Poor wound healing is a source of significant morbidity, with a myriad of psychosocial implications. Research undertaken in our laboratory has focused on wound profiling and therapeutics [2], describing the action of several molecules on the activity of human epidermal cell lines at the wound edge [4,5,6]

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