Specificity of antimicrobial peptide LL-37 to neutralize periodontopathogenic lipopolysaccharide activity in human oral fibroblasts

J Periodontol. 2013 Feb;84(2):256-64. doi: 10.1902/jop.2012.110652. Epub 2012 Mar 23.

Abstract

Background: The antimicrobial peptide LL-37 is known to have a potent lipopolysaccharide (LPS)-neutralizing activity in various cell types. Because of observed heterogeneity within periodontopathogenic LPS, the authors hypothesized that LL-37 had specificity to neutralize such LPS activity. The present study, therefore, aims to investigate the LPS-neutralizing activity of LL-37 to various periodontopathogenic LPS in interleukin-8 (IL-8) production after challenging them in human oral fibroblasts.

Methods: Human periodontal ligament fibroblasts (PDLF) and gingival fibroblasts (GF) were cultured from biopsies of periodontal ligament and gingival tissues. After cell confluence in 24-well plates, LPS (10 μg/mL) from Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans were added with or without LL-37 (10 μg/mL). After 18 hours, the supernatant was collected and analyzed in IL-8 production by enzyme-linked immunosorbent assay.

Results: All periodontopathogenic LPS statistically significantly induced IL-8 production in both PDLF and GF (P <0.01). After neutralization with LL-37, both PDLF and GF showed a statistically significant reduction in IL-8 production compared with LPS-treated groups without LL-37 (P <0.01), and the percentage of reduction in IL-8 production in PDLF appeared to be higher than in GF. In addition, the percentage of reduction in IL-8 production varied considerably according to each periodontopathogenic LPS.

Conclusions: The antimicrobial peptide LL-37 had an ability to suppress periodontopathogenic LPS-induced IL-8 production in both PDLF and GF. Its LPS-neutralizing activity revealed specificity to periodontopathogenic LPS and seemed to be dependent on the heterogeneity within LPS between different genera.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aggregatibacter actinomycetemcomitans / physiology
  • Antimicrobial Cationic Peptides / pharmacology*
  • Cathelicidins
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / physiology
  • Fibroblasts / drug effects*
  • Fusobacterium nucleatum / physiology
  • Gingiva / cytology
  • Gingiva / drug effects*
  • Humans
  • Interleukin-8 / antagonists & inhibitors
  • Limulus Test
  • Lipopolysaccharides / analysis
  • Lipopolysaccharides / antagonists & inhibitors*
  • Periodontal Diseases / microbiology*
  • Periodontal Ligament / cytology
  • Periodontal Ligament / drug effects*
  • Porphyromonas gingivalis / physiology
  • Prevotella intermedia / physiology
  • Species Specificity
  • Young Adult

Substances

  • Antimicrobial Cationic Peptides
  • Interleukin-8
  • Lipopolysaccharides
  • lipopolysaccharide, Escherichia coli O111 B4
  • Cathelicidins