title: Green tea polyphenol epigallocatechin-3-gallate and cranberry proanthocyanidins act in synergy with cathelicidin (LL-37) to reduce the LPS-induced inflammatory response in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts.
abstract: OBJECTIVE
  The human antimicrobial peptide cathelicidin (LL-37) possesses anti-inflammatory properties that may contribute to attenuating the inflammatory process associated with chronic periodontitis. Plant polyphenols, including those from cranberry and green tea, have been reported to reduce inflammatory cytokine secretion by host cells. In the present study, we hypothesized that A-type cranberry proanthocyanidins (AC-PACs) and green tea epigallocatechin-3-gallate (EGCG) act in synergy with LL-37 to reduce the secretion of inflammatory mediators by oral mucosal cells.
  
  
  METHODS
  A three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts treated with non-cytotoxic concentrations of AC-PACs (25 and 50 μg/ml), EGCG (1 and 5 μg/ml), and LL-37 (0.1 and 0.2 μM) individually and in combination (AC-PACs+LL-37 and EGCG+LL-37) were stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). Multiplex ELISA assays were used to quantify the secretion of 54 host factors, including chemokines, cytokines, growth factors, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs).
  
  
  RESULTS
  LL-37, AC-PACs, and EGCG, individually or in combination, had no effect on the regulation of MMP and TIMP secretion but inhibited the secretion of several cytokines. AC-PACs and LL-37 acted in synergy to reduce the secretion of CXC-chemokine ligand 1 (GRO-α), granulocyte colony-stimulating factor (G-CSF), and interleukin-6 (IL-6), and had an additive effect on reducing the secretion of interleukin-8 (IL-8), interferon-γ inducible protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) in response to LPS stimulation. EGCG and LL-37 acted in synergy to reduce the secretion of GRO-α, G-CSF, IL-6, IL-8, and IP-10, and had an additive effect on MCP-1 secretion.
  
  
  CONCLUSIONS
  The combination of LL-37 and natural polyphenols from cranberry and green tea acted in synergy to reduce the secretion of several cytokines by an LPS-stimulated 3D co-culture model of oral mucosal cells. Such combinations show promising results as potential adjunctive therapies for treating inflammatory periodontitis.
authors: Department of Oral Diagnosis and Surgery, Araraquara Dental School, State University of São Paulo, São Paulo, Brazil.
authors: Telma Blanca
authors: TB
authors: Lombardo Bedran
authors: Department of Physiology and Pathology, Araraquara Dental School, State University of São Paulo, São Paulo, Brazil.
authors: Denise
authors: D
authors: Palomari Spolidorio
authors: Oral Ecology Research Group, Faculty of Dentistry, Université Laval, Quebec City, QC, Canada. Electronic address: Daniel.Grenier@greb.ulaval.ca.
authors: Daniel
authors: D
authors: Grenier
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D018920:Coculture Techniques; D004357:Drug Synergism; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005881:Gingiva / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D008070:Lipopolysaccharides; D010936:Plant Extracts / Q000494:pharmacology*; D044945:Proanthocyanidins / Q000494:pharmacology*; D013662:Tea; D029799:Vaccinium macrocarpon; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D010936:Plant Extracts; D044945:Proanthocyanidins; D013662:Tea; D002392:Catechin; C045651:epigallocatechin gallate; D054804:Cathelicidins
keywords: Cathelicidin; Cranberry; Epithelial cell; Fibroblast; Green tea; Periodontal disease