Effect of growth factors on antimicrobial peptides and pro-inflammatory mediators during wound healing
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Antimicrobial peptides (AMPs), such as human beta-defensin-2 (hBD-2) and the CC-chemokine ligand 20 (CCL20), exhibit direct microbicidal effects and mediator-like activity. It was hypothesized that wounding induces the expression of AMPs and pro-inflammatory mediators and that endogenous mediators, such as insulin-like growth factor-1 (IGF-1) and transforming growth factor-alpha (TGF-alpha), modulate this induced expression.
Material and methods
Monolayers of gingival epithelial cells (GECs) and gingival fibroblast (HGFs) from three different donors were wounded using the scratch assay (in vitro wounding) in the presence (test group) or absence (control group) of IGF-1 and TGF-alpha. In vitro wound closure was monitored over time (0, 6, 24, 48, 72 h), and wound areas were microscopically analyzed (Axio-Vision® Software, Zeiss). Gene expression analysis of the GAPDH, hBD-2, CCL20, interleukin-1 beta (IL-1 beta), and interleukin-8 (IL-8) was performed by qPCR.
In comparison to control cells, IGF-1 and TGF-alpha significantly enhanced in vitro wound closure (P < 0.05). In GECs, IGF-1 induced the gene expression of IL-1 beta and IL-8 when compared to control cells (P < 0.05). In HGFs, wounding per se induced the messenger RNA of hBD-2, CCL20, and IL-1 beta, whereas IGF-1 and TGF-alpha reversed this effect (P < 0.05).
In gingival cells, the gene expression of AMPs was altered by injury, and endogenous growth factors further influenced the expression profiles, but with high interindividual differences.
KeywordsAntimicrobial peptides Gingival epithelial cells Gingival fibroblasts IGF-1 In vitro wounding TGF-alpha
The authors kindly thank Mrs. J. Eich and Ms. D. Lalaouni for their excellent technical assistance. This study was financially supported by the German Society of Periodontology (Deutsche Gesellschaft für Parodontologie, DGP; authors: H.D., S.J.) and the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG, Clinical Research Unit 208; authors: TP2, H.D., S.J.; TP7, W.G.; TP4, J.D.; TP8, A.J.; TP9, L.H.; TP10, J.W.).
Conflict of interest
The authors declare no conflict of interest.
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