Effects of cranberry components on human aggressive periodontitis gingival fibroblastsby D. A. Tipton, J. P. Babu, M. Kh. Dabbous

Journal of Periodontal Research

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Year
2012
DOI
10.1111/jre.12023
Subject
Periodontics

Text

Effects of cranberry components on human aggressive periodontitis gingival fibroblasts

Tipton DA, Babu JP, Dabbous MKh. Effects of cranberry components on human aggressive periodontitis gingival fibroblasts. J Periodont Res 2013; 48: 433–442. © 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

Background and 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 andMMP-3, via transcription factors (i.e. NF-jB). Cranberry (Vacciniummacrocarpon) 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-jB activation and IL-6 andMMP-3 production.

Material and Methods: AgP fibroblasts were incubated  6 d with high molecular weight non-dialyzable material (NDM) (derived from cranberry juice (1– 500 lg/mL) or lipopolysaccharide (1 lg/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-jB p65 was measured via binding to an oligonucleotide containing the NF-jB 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 lg/mL) or lipopolysaccharide ± NDM had no effect on viability  7 d exposure. NDM (50 lg/mL) inhibited lipopolysaccharide-stimulated p65 (P  0.003) and constitutive or lipopolysaccharidestimulated 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).

Conclusion: Lack of toxicity of low NDM concentrations, and its inhibition of

NF-jB 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.

D. A. Tipton1,2, J. P. Babu1,2,

M. Kh. Dabbous1,2,3,4 1College of Dentistry, The University of

Tennessee Health Science Center, 62 South

Dunlap Street, Memphis, TN, 38163, USA, 2Department of Bioscience Research, The

University of Tennessee Health Science

Center, 62 South Dunlap Street, Memphis, TN, 38163, USA, 3College of Medicine, The

University of Tennessee Health Science

Center, 62 South Dunlap Street, Memphis, TN, 38163, USA and 4Department of Microbiology,

Immunology and Biochemistry, The University of Tennessee Health Science Center, 62 South

Dunlap Street, Memphis, TN, 38163, USA

David A. Tipton, DDS, PhD, Department of

Bioscience Research, College of Dentistry,

University of Tennessee Health Science

Center, 711 Jefferson Avenue, Rm. 429,

Memphis, TN 38163, USA

Tel: +901-448-6167

Fax: +901-448-7860 e-mail:dtipton@uthsc.edu

Key words: aggressive periodontitis; cranberry; cytokine; fibroblast; lipopolysaccharide; matrix metalloproteinases

Accepted for publication September 9, 2012

J Periodont Res 2013; 48: 433–442

All rights reserved © 2012 John Wiley & Sons A/S.

Published by John Wiley & Sons Ltd

JOURNAL OF PERIODONTAL RESEARCH doi:10.1111/jre.12023

Aggressive periodontitis (AgP) is characterized by generalized or localized extreme periodontal destruction, which involves pro-inflammatory cytokines, including interleukin (IL)-1b and IL-6 (1–3). IL-6 stimulates bone resorption, possibly through stimulating osteoclast formation (4,5). Gingival fibroblasts stimulated by periodontopathogen lipopolysaccharide produce cytokines such as IL-6, and play an important role in local inflammation. The expression of IL-6 is increased at periodontally diseased sites, including gingival soft tissue (4,6,7). Earlier work in this laboratory showed that AgP gingival fibroblasts produce greater amounts of

IL-6 than normal human gingival fibroblasts, and that IL-6 production by these cells is upregulated by lipopolysaccharide (and IL-1b) (8,9).

AgP fibroblasts also produce matrix metalloproteinases (MMPs), including MMP-3, which degrades many extracellular matrix components. A study in this laboratory suggested that human AgP gingival fibroblasts might contribute to periodontal tissue destruction via increased production of MMP-3 (10).

MMP-3 in gingiva is associated with periodontitis, and it may be a key enzyme causing tissue destruction in other inflammatory diseases, including arthritis (11,12).

Transcription of the IL-6 and

MMP-3 genes (and a large number of other genes involved in inflammation and tissue damage) are regulated by the transcription factor nuclear factor-kappa B (NF-jB) (13). Work in this laboratory and others showed that cyclo-oxygenase-2 inhibitors inhibit NF-jB activation (14), which suggested that they might be useful in treating periodontal inflammation.

However, cyclo-oxygenase-2 inhibitors have been linked to heart and renal failure, limiting their use as antiinflammatory agents (15,16). These findings have led to renewed attention to natural products potentially useful in treating the inflammation and tissue destruction associated with periodontal diseases (17,18).

The focus of this research was the cranberry (Vaccinium macrocarpon), which contains polyphenolic compounds, particularly proanthocyanidins with A-type linkages. Cranberry components have long been used to promote human health, and have the ability to prevent urinary tract infections, reduce salivary bacterial levels, and inhibit biofilm formation, among many other effects (19–21).