The Antimicrobial Peptide Human Beta-Defensin-3 Is Induced by Platelet-Released Growth Factors in Primary Keratinocytes.

Andreas Bayer,Sebastian Lippross,Franziska Rademacher,Mersedeh Tohidnezhad,Thomas Pufe,Jochen Cremer,Tim Klüter,Jürgen Harder,Justus Lammel,Regine Gläser,Peter Behrendt,Holger Jahr

doi:10.1155/2017/6157491

Abstract

Platelet-released growth factors (PRGF) and its related clinically used formulations (e.g., Vivostat Platelet-Rich Fibrin (PRF®)) contain a variety of chemokines, cytokines, and growth factors and are therefore used to support healing of chronic, hard-to-heal, or infected wounds. Human beta-defensin-3 (hBD-3) is an antimicrobial peptide inducibly expressed in human keratinocytes especially upon wounding. The potent antimicrobial activity of hBD-3 together with its wound closure-promoting activities suggests that hBD-3 may play a crucial role in wound healing. Therefore, we analyzed the influence of PRGF on hBD-3 expression in human primary keratinocytes in vitro. In addition, we investigated the influence of Vivostat PRF on hBD-3 expression in artificially generated human skin wounds in vivo. PRGF treatment of primary keratinocytes induced a significant, concentration- and time-dependent increase in hBD-3 gene expression which was partially mediated by the epidermal growth factor receptor (EGFR). In line with these cell culture data, in vivo experiments revealed an enhanced hBD-3 expression in experimentally produced human wounds after the treatment with Vivostat PRF. Thus, the induction of hBD-3 may contribute to the beneficial effects of thrombocyte concentrate lysates in the treatment of chronic or infected wounds.

Highlights

Platelet-released growth factors (PRGF) is a thrombocyte concentrate lysate comprising a multitude of chemokines, cytokines, and growth factors [1,2,3,4,5]
We analyzed Human beta-defensin-3 (hBD-3) gene expression after 4, 12, 24, 48, and 72 hours of PRGF stimulation to assess the time kinetic of PRGFmediated human beta-defensin- (hBD-)3 induction. hBD-3 was significantly induced in primary keratinocytes after 24 hours of PRGF stimulation (Figure 2)
Since we have previously shown that PRGF rapidly induced IL-6 gene expression in primary human keratinocytes [17], we investigated the possible role of IL-6 on the PRGF-mediated hBD-3 induction in human keratinocytes by blocking the IL-6 receptor with the IL-6 receptor-neutralizing antibody tocilizumab

Summary

Introduction

Platelet-released growth factors (PRGF) is a thrombocyte concentrate lysate comprising a multitude of chemokines, cytokines, and growth factors [1,2,3,4,5]. Its clinically related formulation Vivostat PRF has already been used successfully to support healing of patients’ hard-to-heal wounds in vivo [13]. One possible reason for a malfunctioning wound healing process of the skin is a local wound infection with potential pathogenic bacterial species. In this situation, human keratinocytes establish a chemical defense system based on the production of antimicrobial peptides, for example, human beta-defensin- (hBD-) 2 and hBD-3 to defeat these microbial threats [14,15,16]. We demonstrated that PRGF induces the antimicrobial peptide hBD-2 in Mediators of Inflammation primary keratinocytes [17] showing for the first time that PRGF is able to induce the expression of antimicrobial peptides in cultured keratinocytes as well as in wounded skin

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Conclusion