Wound healing potential of a dimeric InlB variant analyzed by in vitro experiments on re-epithelialization of human skin models |
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| Institution: | 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;1. Center of Experimental and Applied Cutaneous Physiology, Department of Dermatology, Charité – Universitätsmedizin Berlin, Germany;2. Zebet at Federal Institute of Risk Assessment (BfR), Berlin, Germany;3. Department of Pharmaceutical Technology, Friedrich-Schiller-University Jena, Jena, Germany;4. Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense, Denmark;5. Department of Surgery, Charité – Universitätsmedizin Berlin, Germany;6. Department of Chemistry, Humboldt-Universität zu Berlin, Berlin, Germany;7. Institute of Pharmacy (Pharmacology and Toxicology), Freie Universität Berlin, Germany;1. Department of Chemistry and Applied Biosciences, ETH Zürich, Zürich, Switzerland;1. Department of Dermatology, Jichi Medical University, Tochigi, Japan;2. Department of Biochemistry, Jichi Medical University, Tochigi, Japan;3. JADECOM (Japan Association for Development for Community Medicine), Tochigi, Japan;1. Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin—Madison, Madison, WI, USA;2. Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin—Madison, Madison, WI, USA |
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| 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. |
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| Keywords: | 3D skin model Hydrogel Internalin B Keratinocyte MET receptor Migration MTT assay Proliferation Scratch assay Wound healing |
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