Baicalin Downregulates Porphyromonas gingivalis Lipopolysaccharide-Upregulated IL-6 and IL-8 Expression in Human Oral Keratinocytes by Negative Regulation of TLR Signaling

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Systems biology unravels novel biomarkers in heterogeneous PgLPS-HGF interaction

Session - Markers of Periodontal Pathogenesis: abstract no. 2983OBJECTIVES: Porphyromonas gingivalis lipopolysaccharide (PgLPS) is a crucial virulence factor strongly associated with chronic periodontitis. It displays significant amount of lipid A structural heterogeneity, containing both tetra- (LPS1435/1449) and penta-acylated (LPS1690) isoforms. We recently showed these isoforms evoked an opposing immuno-inflammatory response in human gingival fibroblasts (HGFs) (IADR/Barcelona, 2010). In the present study we have for the first time employed a systems biology approach using advanced tools in transcriptomics, proteomics, metabolomics and bioinformatics to determine novel molecules induced in the PgLPS-HGF Interaction. METHODS: Primary HGFs were stimulated with 1 µg/ml PgLPS isoforms for 24h using E. coli LPS as the reference. Cellular proteins were separated by 2D-PAGE coupled with quantitative fluorescent dyes. Differentially expressed proteins ≥ 2 folds (P < 0.05) were identified by tandem mass spectrometry (MS/MS). Tryptic digested secretory proteins were identified by affinity chromatography combined Nano-LC-MALDI TOF/TOF MS/MS. Generic and specific biomarkers were cataloged using bioinformatics pathways. HGF RNA was subjected to transcriptomic analysis using toll-like receptor pathway gene-arrays. RESULTS: Overall, 32 differentially expressed cell-bound proteins and 176 secretory proteins (metabolome) were identified in HGF by using LPS1435/1449 and LPS1690. Both induced the expression of cathepsins, osteonectin, ADAM-28 and TGF-β binding protein. Conversely, PgLPS1690 induced the expression of IL-6, IL-8, GM-CSF, GCSF and hitherto underscribed small molecular weight proteins. In contrast, PgLPS1435/1449 induced the expression of anti-inflammatory molecules such as SOCS box proteins, IL-20, TNF-α inhibitor and down-regulated the expression of lipid peroxidation pathway molecules perilipin, superoxide dismutase, peroxiredoxin-6 and thioredoxin. PgLPS1690 activated the apoptosis-related pathway (Ras-Bax), whereas Pg LPS1435/1449 inhibited it. CONCLUSION: The present study unraveled novel biomarkers with immuno-inflammatory agonist-antagonist activities released by HGFs in their interaction with the heterogeneous PgLPS isoforms, which may lay the foundation for the development of novel diagnostic and therapeutic approaches to control periodontal diseases (GRF HKU766909M to LJ JIN).link_to_OA_fulltextThe 89th General Session and Exhibition of IADR/AADR/CADR, San Diego, CA., 16-19 March 2011

Porphyromonas gingivalis LPS modulates immuno-inflammatory response in human gingival fibroblasts

Objectives: P. gingivalis is a major periodontopathogens. P. gingivalis lipopolysaccharide (LPS) is highly heterogeneous and it differentially modulates immuno-inflammatory responses. This study was to examine the effects of different isoforms of P. gingivalis LPS on the expression of pro-inflammatory cytokines and MMPs in human gingival fibroblasts (HGF)...The International Association for Dental Research (IADR) 88th General Session and Exhibition 2010, Barcelona, Spain, 14-17 July 2010. In Journal of Dental Research, 2010, v. 89 n. Spec Iss B, Abstract no. 342

Novel pathogenic mechanisms of heterogeneous Pg-LPS structures in periodontal disease

IADR/Unilever Hatton Awards 245 - IADR/Unilever Hatton Awards - Senior Category: Basic Science (Poster Session): no. 2643OBJECTIVE: Porphyromonas gingivalis lipopolysaccharide (PgLPS) displays significant amount of lipid-A heterogeneity, containing both tetra (LPS1435/1449) and penta-acylated (LPS1690) isoforms. We recently demonstrated PgLPS isoforms differentially modulate the immuno-inflammatory response in human gingi...link_to_OA_fulltex

Prunus mume extract exhibits antimicrobial activity against pathogenic oral bacteria

Objectives. Prunus mume is a common fruit in Asia, which has been used in traditional Chinese medicine. In this study, we focused on the antimicrobial properties of Prunus mume extract against oral pathogens related to dental caries and periodontal diseases. Study design. A total of 15 oral pathogens including Streptococcus mutans, S. sobrinus, S. mitis, S. sanguinis, Lactobacillus acidophilus, P. gingivalis, Aggregatibacter actinomycetemcomitans, and Candida species were included in the study. Initially, agar diffusion assay was performed to screen the antimicrobial activities of Prunus mume extract. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for sensitive species. Effect of Prunus mume extract on human oral keratinocytes (HOK) viability was also tested. Result. In the agar diffusion assay, drug suspension of 2g/mL was able to inhibit all the bacterial species tested, but not the fungal species. MIC and MBC range of Prunus mume extract against the oral bacteria was 0.15625-0.0003g/mL and P. gingivalis being the most susceptible species. Prune extract did not cause any detrimental effect on HOK. Conclusion. Prunus mume extract may be a potential candidate for developing an oral antimicrobial agent to control or prevent dental diseases associated with oral pathogenic bacteria. © 2011 The Authors. International Journal of Paediatric Dentistry © 2011 BSPD, IAPD and Blackwell Publishing Ltd.link_to_subscribed_fulltex