Nawarat Wara-aswapati

CONCISE REVIEW Biological: Interleukin 1 Signal Transduction— Current Concepts and Relevance to Periodontitis

This review examines a well-characterized factor, interleukin 1 (IL-1), that has recently received considerable attention. A level of understanding is emerging that goes beyond simple recognition that IL-1 plays a role in disease, and begins to explain the molecular mechanisms of function. This review summarizes some current information on the importance of IL-1 in periodontitis as well as the signal transduction of IL-1, from binding to its cell-surface receptors, to the activation of cytoplasmic mediators and transcription factors responsible for the induction of target genes. The effect of IL-1 signal transduction is ultimately the activation and repression of specific transcription factors that regulate genes responsible for cellular activities. As additional steps of signal transduction become better-characterized, these insights may facilitate the development of improved therapeutic approaches for controlling inflammation and connective tissue destruction in a variety of diseases.

Current and future periodontal tissue engineering

Periodontitis is an inflammatory disease that leads to the loss of tooth-supporting tissues. Conventional periodontal treatment is generally unable to promote regeneration of the damaged periodontal structures. Recently, several studies have investigated the use of tissue engineering to facilitate predictable periodontal regeneration. This article reviews various technologies related to periodontal tissue engineering. These include bone grafting, guided tissue regeneration, enamel matrix protein derivative, growth factors, stem cell therapy, cell sheet engineering and laser treatment. Studies carried out by this group, and available clinical data, together with the authors own clinical experiences, are discussed. In addition, possible new directions that need to be exploited to make periodontal tissue engineering a clinical success are discussed herein.

TRAF6 activation of PI 3-kinase-dependent cytoskeletal changes is cooperative with Ras and is mediated by an interaction with cytoplasmic Src.

Interleukin 1 (IL-1) has been implicated in the reorganization of the actin cytoskeleton. An expression vector encoding a PKB/Akt pleckstrin-homology domain fused to a fluorescent protein was used to detect phosphoinositide 3-kinase (PI 3-kinase) products. It was observed that PI 3-kinase was activated either by treatment with IL-1 or by expression of either TRAF6, Src, MyD88 or dominant-positive PI 3-kinase, and resulted in the formation of long filopodia-like cellular protrusions that appeared to branch at membrane sites consisting of clusters of phosphoinositide. This depended upon a TRAF6 polyproline motif and Src catalytic activity, and was blocked by inhibitors of PI 3-kinase, Src and Ras. Using both conventional and split fluorescent protein probes fused to expressed TRAF6 and Src in living cells, the polyproline sequence of TRAF6 and the Src-homology 3 (SH3) domain of Src were shown to be required for interaction between these two proteins. Interaction occurred within the cytoplasm, and not at either the cell membrane or cytoplasmic sequestosomes. In addition, co-transfection of vectors expressing fluorescent-protein-fused TRAF6 and non-fluorescent MyD88, IRAK1 and IRAK2 revealed an inverse correlation between increased sequestosome formation and activation of both PI 3-kinase and NF-κB. Although a key factor in TRAF6-dependent activation of PI 3-kinase, ectopic expression of Src was insufficient for NF-κB activation and, in contrast to NF-κB, was not inhibited by IRAK2.

Red bacterial complex is associated with the severity of chronic periodontitis in a Thai population.

BACKGROUND The distribution of periodontal pathogens differs in various geographic locations and racial/ethnic groups. This study investigated the microbiological features of chronic periodontitis (CP) patients in Thailand. METHODS Subgingival plaque samples from 20 non-periodontitis subjects, 20 patients with mild CP, and 20 patients with moderate to severe CP were examined using polymerase chain reaction (PCR) to identify Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans. RESULTS In the moderate to severe CP patients, there was high prevalence of P. gingivalis (95%), T. forsythia (95%), T. denticola (80%), as well as the red complex (coexistence of all three species at the same lesion) (75%). A. actinomycetemcomitans was detected in only 35% of the patients in this study group. P. gingivalis was detected in as high as 45% of the non-periodontitis controls. CP and disease severity were significantly related to the presence of T. forsythia together with T. denticola and the red complex. The red complex was not found in any non-periodontitis site. CONCLUSION Red complex bacteria were predominant periodontal pathogens of the moderate to severe form of CP in this Thai population. The presence of T. forsythia together with T. denticola, and the red complex species at the same site were significantly associated with the disease severity.

Induction of toll-like receptor expression by Porphyromonas gingivalis.

BACKGROUND Toll-like receptors (TLRs) play pivotal roles in host immune responses and have been suggested to be involved in the development of many infectious diseases. In this study, the mRNA expression levels of TLR2, TLR4, and TLR9 and their relationship with periodontopathic bacteria in periodontal tissue are examined. Furthermore, the mechanism of TLR induction by Porphyromonas gingivalis is investigated in human gingival fibroblasts (HGFs). METHODS Gingival tissue and subgingival plaque samples were collected from 19 patients with chronic periodontitis (CP) and 16 control individuals without periodontitis. Gene expression levels in the tissues and in HGFs were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The numbers of periodontopathic bacteria were determined by quantitative real-time PCR. RESULTS The expression levels of TLR2 and TLR9 were significantly higher in the tissues of patients with CP compared to the tissues of control individuals. The mRNA levels of TLR2 and TLR9, but not TLR4, were positively correlated with the number of P. gingivalis in subgingival plaque. P. gingivalis sonicated extract, P. gingivalis lipopolysaccharide, P. gingivalis DNA, and tumor necrosis factor-α(TNF-α) could significantly upregulate the mRNA expression of TLR2 in HGFs. Furthermore, P. gingivalis-mediated TLR2 expression was suppressed by TNF-α antibody. CONCLUSIONS This study suggests that P. gingivalis infection induces TLR2 and TLR9 upregulation in patients with CP. P. gingivalis-induced TLR2 expression in HGFs is partially dependent on TNF-α and may lead to sensitization of HGFs to bacterial components encountered in the periodontal microenvironment.

Association between periodontitis and anti‐cardiolipin antibodies in Buerger disease

AIM Anti-cardiolipin (CL) antibodies can be induced in Buerger disease (BD), an inflammatory occlusive disorder affecting peripheral blood vessels, in response to bacteria bearing homology to the TLRVYK peptide of a phospholipid-binding plasma protein beta-2-glycoprotein I. TLRVYK homologies are present in Porphyromonas gingivalis (TLRIYT) and Treponema denticola (TLALYK). This study investigated the association between periodontal infection and anti-CL antibodies in BD patients. MATERIAL AND METHODS Periodontal conditions were examined in 19 BD patients and 25 systemically healthy control subjects. All subjects were heavy smokers. Serum anti-CL, anti-TLRVYK, anti-TLRIYT, and anti-TLALYK antibodies were assessed using the enzyme-linked immunosorbent assay. RESULTS BD patients had a significantly higher prevalence of periodontitis, more severe periodontal destruction and increased titres of serum anti-CL, anti-TLRVYK, anti-TLRIYT, and anti-TLALYK antibodies compared with healthy subjects. The levels of anti-CL antibodies positively correlated with those of the three anti-peptide antibodies. Anti-CL antibody titres were significantly associated with the percentage of sites with clinical attachment level >or=4 mm in BD patients. CONCLUSION Elevated anti-CL antibody levels were associated with periodontal destruction in BD patients. Periodontopathic bacteria may serve as exogenous antigens that stimulate the anti-CL antibody production through molecular mimicry between the bacterial peptides and a host plasma protein.

Modulation of Wnt5a Expression by Periodontopathic Bacteria

Wingless proteins, termed Wnt, are involved in embryonic development, blood cell differentiation, and tumorigenesis. In mammalian hematopoiesis, Wnt signaling is essential for stem-cell homeostasis and lymphocyte differentiation. Recent studies have suggested that these molecules are associated with cardiovascular diseases, rheumatoid arthritis, and osteoarthritis. Furthermore, Wnt5a signaling is essential for the general inflammatory response of human macrophages. Periodontitis is a chronic inflammatory disease caused by gram-negative periodontopathic bacteria and the resultant host immune response. Periodontitis is characterized by loss of tooth-supporting structures and alveolar bone resorption. There have been no previous reports on Wnt5a expression in periodontitis tissue, and only few study reported the molecular mechanisms of Wnt5a expression in LPS-stimulated monocytic cells. Using RT-PCR, we demonstrated that Wnt5a mRNA expression was up-regulated in chronic periodontitis tissue as compared to healthy control tissue. P. gingivalis LPS induced Wnt5a mRNA in the human monocytic cell line THP-1 with a peak at 4 hrs after stimulation. P. gingivalis LPS induced higher up-regulation of Wnt5a mRNA than E. coli LPS. The LPS receptors TLR2 and TLR4 were equally expressed on the surface of THP-1 cells. P. gingivalis LPS induced IκBα degradation and was able to increase the NF-κB binding activity to DNA. P. gingivalis LPS-induced Wnt5a expression was inhibited by NF-κB inhibitors, suggesting NF-κB involvement. Furthermore, IFN-γ synergistically enhanced the P. gingivalis LPS-induced production of Wnt5a. Pharmacological investigation and siRNA experiments showed that STAT1 was important for P. gingivalis LPS-induced Wnt5a expression. These results suggest that the modulation of Wnt5a expression by P. gingivalis may play an important role in the periodontal inflammatory process and serve a target for the development of new therapies.

Anti-inflammatory effect of Streblus asper leaf extract in rats and its modulation on inflammation-associated genes expression in RAW 264.7 macrophage cells

AIM OF THIS STUDY Streblus asper is a medicinal plant from Thailand used in folk medicine for the treatment of several inflammatory diseases. In this study, we investigated the anti-inflammatory effect of Streblus asper leaf ethanolic extract (SAE). MATERIALS AND METHODS The experimental carrageenan-induced paw edema in rats was performed in which the SAE at doses of 125, 250, 500 mg/kg body weight was intraperitoneally administered to the rats. Then, reverse transcriptive polymerase chain reaction (RT-PCR) technique was also performed to determine the effect of SAE on the expression of inflammation-associated genes in RAW 264.7 macrophage cells stimulated with lipopolysaccharide (LPS). RESULTS The SAE at all given doses caused a significant dose-dependent inhibition of edema (p<0.05). Moreover, the significant and dose-dependent LPS-induced cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) mRNA expressions were demonstrated in RAW 264.7 cells treated with SAE. The inhibition is selective, since COX-1 mRNA expression did not change in the presence of SAE. CONCLUSION The results of this study are the first scientific evidence on the molecular effects of Streblus asper as a potential anti-inflammatory agent, which supports the fact that the plant is employed in traditional remedies.

Periodontitis and Gestational Diabetes Mellitus in Non-Smoking Females

BACKGROUND Chronic inflammation has been implicated in the pathogenesis of gestational diabetes mellitus (GDM). Periodontal disease is associated with increased levels of inflammatory mediators and may be a risk factor for GDM. The authors aimed to examine the association between periodontitis and GDM among non-smoking pregnant females. METHODS This case-control study included 50 females who were diagnosed with GDM and 50 age- and hospital-matched females without diabetes in Khon Kaen, Thailand. Full-mouth periodontal examinations were performed during pregnancy by two calibrated dentists who were unaware of the case-control status. Periodontitis was defined as ≥1 site with probing depth (PD) ≥5 mm and clinical attachment level (CAL) ≥2 mm at the same site. Serum samples were collected to measure C-reactive protein (CRP), tumor necrosis factor-α, and interleukin-6 levels. Analyses were performed using conditional logistic regression. RESULTS Fifty percent of the case females had periodontitis compared to 26% of the controls. Females with GDM had significantly higher mean PD and CAL, more sites with bleeding on probing, and increased levels of CRP compared to the controls. Periodontitis was significantly associated with GDM (odds ratio = 3.00, 95% confidence interval = 1.19 to 7.56). The association remained significant with additional adjustment for family history of diabetes, prepregnancy body mass index, and weight gain during pregnancy. CONCLUSIONS The results suggest that periodontitis is associated with GDM. Therefore, clinicians should assess periodontal conditions of pregnant females.

Conserved ETS Domain Arginines Mediate DNA Binding, Nuclear Localization, and a Novel Mode of bZIP Interaction

The DNA-binding ETS transcription factor Spi-1/PU.1 is of central importance in determining the myeloid-erythroid developmental switch and is required for monocyte and osteoclast differentiation. Many monocyte genes are dependent upon this factor, including the gene that codes for interleukin-1β. It has long been known that the conserved ETS DNA-binding domain of Spi-1/PU.1 functionally cooperates via direct association with a diverse collection of DNA-binding proteins, including members of the basic leucine zipper domain (bZIP) family. However, the molecular basis for this interaction has long been elusive. Using a combination of approaches, we have mapped a single residue on the surface of the ETS domain critical for protein tethering by the C/EBPβ carboxylterminal bZIP domain. This residue is also important for nuclear localization and DNA binding. In addition, dependence upon the leucine zipper suggests a novel mode for both protein-DNA interaction and functional cooperativity.