title: Effects of cranberry components on human aggressive periodontitis gingival fibroblasts.
abstract: OBJECTIVE
  Aggressive periodontitis (AgP) causes rapid periodontal breakdown involving AgP gingival fibroblast production of cytokines [i.e. interleukin (IL)-6, a bone metabolism regulator], and matrix metalloproteinase (MMP)-3. Lipopolysaccharide upregulates fibroblast IL-6 and MMP-3, via transcription factors (i.e. NF-κB). Cranberry (Vaccinium macrocarpon) inhibits lipopolysaccharide-stimulated macrophage and normal gingival fibroblast activities, but little is known of its effects on AgP fibroblasts. Objectives of this study are to use AgP fibroblasts, to determine cytotoxicity of cranberry components or periodontopathogen (Fusobacterium nucleatum, Porphyromonas gingivalis) lipopolysaccharide ± cranberry components, and effects of cranberry components on lipopolysaccharide-stimulated NF-κB activation and IL-6 and MMP-3 production.
  
  
  METHODS
  AgP fibroblasts were incubated ≤ 6 d with high molecular weight non-dialyzable material (NDM) (derived from cranberry juice (1-500 μg/mL) or lipopolysaccharide (1 μg/mL) ± NDM. Membrane damage and viability were assessed by enzyme activity released into cell supernatants and activity of a mitochondrial enzyme, respectively. Secreted IL-6 and MMP-3 were measured by ELISA. NF-κB p65 was measured via binding to an oligonucleotide containing the NF-κB consensus site. Data were analyzed using analysis of variance and Scheffe's F procedure for post hoc comparisons.
  
  
  RESULTS
  Short-term exposure to NDM, or lipopolysaccharide ± NDM caused no membrane damage. NDM (≤ 100 μg/mL) or lipopolysaccharide ± NDM had no effect on viability ≤ 7 d exposure. NDM (50 μg/mL) inhibited lipopolysaccharide-stimulated p65 (P ≤ 0.003) and constitutive or lipopolysaccharide-stimulated MMP-3 (P ≤ 0.02). NDM increased AgP fibroblast constitutive or lipopolysaccharide-stimulated IL-6 (P ≤ 0.0001), but inhibited normal human gingival fibroblast IL-6 (P ≤ 0.01).
  
  
  CONCLUSIONS
  Lack of toxicity of low NDM concentrations, and its inhibition of NF-κB and MMP-3, suggest that cranberry components may regulate AgP fibroblast inflammatory responses. Distinct effects of NDM on AgP and gingival fibroblast production of IL-6 (which can have both positive and negative effects on bone metabolism) may reflect phenotypic differences in IL-6 regulation in the two cell types.
authors.affiliation: Department of Bioscience Research, College of Dentistry, The University of Tennessee Health Science Center, Memphis, TN 38163, USA. dtipton@uthsc.edu
authors.forename: D A
authors.initials: DA
authors.lastname: Tipton
authors.forename: J P
authors.initials: JP
authors.lastname: Babu
authors.forename: M Kh
authors.initials: MKh
authors.lastname: Dabbous
mesh_terms: D010520:Aggressive Periodontitis / Q000276:immunology / Q000473:pathology*; D000872:Anthocyanins / Q000494:pharmacology; D018929:Cell Culture Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects* / Q000276:immunology; D016967:Fusobacterium nucleatum / Q000502:physiology; D005881:Gingiva / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D019278:Matrix Metalloproteinase 3 / Q000032:analysis / Q000187:drug effects; D016328:NF-kappa B / Q000187:drug effects; D010936:Plant Extracts / Q000627:therapeutic use*; D016966:Porphyromonas gingivalis / Q000502:physiology; D044945:Proanthocyanidins / Q000494:pharmacology; D013997:Time Factors; D051996:Transcription Factor RelA / Q000032:analysis / Q000187:drug effects; D029799:Vaccinium macrocarpon
chemical_list: D000872:Anthocyanins; C508600:IL6 protein, human; D015850:Interleukin-6; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010936:Plant Extracts; D044945:Proanthocyanidins; D051996:Transcription Factor RelA; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3