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Eur J Pharm Biopharm. 2014 Feb;86(2):277-83. doi: 10.1016/j.ejpb.2013.10.003. Epub 2013 Oct 15.

Wound healing potential of a dimeric InlB variant analyzed by in vitro experiments on re-epithelialization of human skin models.

Author information

1
Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany.
2
Department of Molecular Structural Biology, Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany.
3
Department of Chemistry and Center for Biotechnology (CeBiTec), Bielefeld University, Bielefeld, Germany.
4
Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany. Electronic address: C.Mueller-Goymann@tu-braunschweig.de.

Abstract

A constitutively dimeric truncated variant of internalin B (InlB321-CD), acting as stimulator of the receptor tyrosine kinase MET, was tested for dermal wound-healing potential. Due to a lack of the endogenous MET agonist HGF/SF in chronic wounds, HGF/SF substitution by an InlB321-CD-loaded hydrogel might be beneficial in chronic wound therapy. In this study, InlB321-CD in solution and incorporated in a hydrogel was tested for mitogenic effects on immortalized human dermal keratinocytes (HaCaT) with an MTT assay. Cell migration was investigated with a scratch assay on primary keratinocytes (PHK) and on HaCaT. For the latter, scratching needed to be mitomycin C-controlled. InlB321-CD effects on a model of human skin were analyzed histologically with respect to viability. InlB321-CD led to dose-dependent proliferative effects on HaCaT cells whereas the equimolar dose of monomeric InlB321 did not. Upon hydrogel incorporation of InlB321-CD its mitogenic activity for HaCaT cells was maintained thus confirming the hydrogel as a promising drug delivery system. Motogenic effects were shown on both HaCaT and PHK cells. InlB321-CD neither possesses cytotoxic effects on the viability of a human skin model nor alters its organotypic cell morphology.

KEYWORDS:

3D skin model; Hydrogel; Internalin B; Keratinocyte; MET receptor; MTT assay; Migration; Proliferation; Scratch assay; Wound healing

PMID:
24140590
DOI:
10.1016/j.ejpb.2013.10.003
[Indexed for MEDLINE]
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