Bee-derived antibacterial peptide, defensin-1, promotes wound reepithelialisation <i>in vitro</i> and <i>in vivo</i>

  • M Bucekova Slovak Academy of Sciences
  • Martin Sojka Slovak Medical University
  • Ivana Valachova Comenius University
  • Simona Martinotti University of Piemonte Orientale
  • Elia Ranzato University of Piemonte Orientale
  • Zoltan Szep Cytopahtos Laboratory
  • Viktor Majtan Slovak Medical University
  • Jaroslav Klaudiny Slovak Academy of Sciences
  • Juraj Majtan Slovak Academy of Sciences

Abstract

Royal jelly (RJ) has successfully been used as a remedy in wound healing. RJ has multiple effects, including antibacterial, anti-inflammatory and immunomodulatory activities, in various cell types. However, no component(s) (other than antibacterial) have been identified in RJ-accelerated wound healing. In this study, we demonstrate that keratinocytes are responsible for the elevated production of matrix metalloproteinase-9 (MMP-9) after incubation with a water extract of RJ. Furthermore, the keratinocyte migration and wound closure rates were significantly increased in the presence of RJ extract. MMP-9 production was reduced significantly following proteinase K treatment but remained stable after heat treatment, indicating that active component(s) have a proteinous character. To identify the component responsible for inducing MMP-9 production, RJ extract was fractionated using C18 RP-HPLC. In fractions exhibiting stimulatory activity, we immunochemically detected the bee-derived antibacterial peptide, defensin-1. Defensin-1 was cloned, and recombinant peptide was produced in a baculoviral expression system. Defensin-1 stimulated MMP-9 secretion from keratinocytes and increased keratinocyte migration and wound closure in vitro. In addition, defensin-1 promoted re-epithelisation and wound closure in uninfected excision wounds. These data indisputably demonstrate that defensin-1, a regular but concentration variable factor found in honey and RJ, contributes to cutaneous wound closure by enhancing keratinocyte migration and MMP-9 secretion.

Author Biographies

M Bucekova, Slovak Academy of Sciences
Laboratory of Apidology and Apitherapy Institute of Molecular Biology Slovak Academy of Sciences Bratislava; and Institute of Zoology Slovak Academy of Sciences
Martin Sojka, Slovak Medical University
Department of Microbiology Faculty of Medicine Slovak Medical University
Ivana Valachova, Comenius University
Faculty of Natural Sciences Comenius University
Simona Martinotti, University of Piemonte Orientale
DiSIT-Dipartimento di Scienze e Innovazione Tecnologica University of Piemonte Orientale
Elia Ranzato, University of Piemonte Orientale
DiSIT-Dipartimento di Scienze e Innovazione Tecnologica University of Piemonte Orientale
Zoltan Szep, Cytopahtos Laboratory
Cytopahtos Laboratory
Viktor Majtan, Slovak Medical University
Department of Microbiology Faculty of Medicine Slovak Medical University
Jaroslav Klaudiny, Slovak Academy of Sciences
Institute of Chemistry Slovak Academy of Sciences
Juraj Majtan, Slovak Academy of Sciences
Laboratory of Apidology and Apitherapy Institute of Molecular Biology Slovak Academy of Sciences; Institute of Zoology Slovak Academy of Sciences; and Department of Microbiology Faculty of Medicine Slovak Medical University
Section
Wound Care