title: Synergistic anti-inflammatory activity of the antimicrobial peptides human beta-defensin-3 (hBD-3) and cathelicidin (LL-37) in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts.
abstract: Given the spread of antibiotic resistance in bacterial pathogens, antimicrobial peptides that can also modulate the immune response may be a novel approach for effectively controlling periodontal infections. In the present study, we used a three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) to investigate the anti-inflammatory properties of human beta-defensin-3 (hBD-3) and cathelicidin (LL-37) and to determine whether these antimicrobial peptides can act in synergy. The 3D co-culture model composed of gingival fibroblasts embedded in a collagen matrix overlaid with gingival epithelial cells had a synergistic effect with respect to the secretion of IL-6 and IL-8 in response to LPS stimulation compared to fibroblasts and epithelial cells alone. The 3D co-culture model was stimulated with non-cytotoxic concentrations of hBD-3 (10 and 20 µM) and LL-37 (0.1 and 0.2 µM) individually and in combination in the presence of A. actinomycetemcomitans LPS. A multiplex ELISA assay was used to quantify the secretion of 41 different cytokines. hBD-3 and LL-37 acted in synergy to reduce the secretion of GRO-alpha, G-CSF, IP-10, IL-6, and MCP-1, but only had an additive effect on reducing the secretion of IL-8 in response to A. actinomycetemcomitans LPS stimulation. The present study showed that hBD-3 acted in synergy with LL-37 to reduce the secretion of cytokines by an LPS-stimulated 3D model of gingival mucosa. This combination of antimicrobial peptides thus shows promising potential as an adjunctive therapy for treating inflammatory periodontitis.
authors.affiliation: Department of Oral Diagnosis and Surgery, Araraquara Dental School, State University of São Paulo, São Paulo, Brazil.
authors.forename: Telma Blanca Lombardo
authors.initials: TB
authors.lastname: Bedran
authors.affiliation: Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Márcia Pinto Alves
authors.initials: MP
authors.lastname: Mayer
authors.affiliation: Department of Physiology and Pathology, Araraquara Dental School, State University of São Paulo, São Paulo, Brazil.
authors.forename: Denise Palomari
authors.initials: DP
authors.lastname: Spolidorio
authors.affiliation: Oral Ecology Research Group, Faculty of Dentistry, Université Laval, Quebec City, QC, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Grenier
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000187:drug effects; D000890:Anti-Infective Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D023083:beta-Defensins / Q000494:pharmacology*; D054804:Cathelicidins
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; C422950:DEFB103A protein, human; D023083:beta-Defensins; D054804:Cathelicidins