Platelet responses and anaphylaxis-like shock induced in mice by intravenous injection of whole cells of oral streptococci

Intravenous injection of lyophilized whole cells of various oral streptococcal strains into muramyldipeptide (MDP)-primed C3H/HeN mice induces rapid anaphylactoid shock. Here we examined the mechanism underlying this shock. In non-primed mice, Streptococcus intermedius K-213K (SiK213) and Streptococcus constellatus T21 (ScT21) produced little or no sign of shock. In MDP-primed mice, SiK213 caused lethal shock, while ScT21 only had a weak effect. SiK213 induced decreases in blood platelets and 5-hydroxytryptamine (5HT) preceding the shock, while the effects of ScT21 were weak. The SiK213-induced 5HT decrease and shock were reduced by a complement-C5 inhibitor. These results suggest that (i). streptococcal bacterial cells can induce rapid platelet responses, (ii). complement-dependent degradation of platelets may be involved in streptococcus-induced shock, (iii). the streptococcus-induced platelet degradation or degranulation may occur largely in the systemic circulation, and (iv). platelets may play a role not only in infectious diseases caused by gram-negative bacteria, but also in diseases caused by gram-positive bacteria.     

Comparison of physical properties of polycarboxylate-based and conventional root canal sealers.

Several physical properties of nine commercial root canal sealers were evaluated in vitro and were compared with those of two experimental endodontic materials based on polycarboxylate formulations. Flow, setting time, compressive strength, radiopacity, adhesion to root dentin, and solubility were evaluated. The zinc oxideeugenol root canal sealers were typically of low strength and high solubility. These sealers and the Diaket polyvinyl resin sealer showed no adhesion to dentin. The epoxy resin AH26 showed superior properties with respect to strength, flow, radiopacity, and adhesion; solubility of this material was high. The polycarboxylate formulation showed significantly higher values over the commercial sealers in properties of strength, adhesion, and reduced solubility. The tensile adhesive bond strength of the polycarboxylate to root dentin was twice that of AH26. A wide variation in properties of the commercial materials tested showed the empirical nature of these "sealers." Further testing of polycarboxylate endodontic sealers is indicated.     

5-HT、NPY与TMD患者关节源性疼痛关系的研究

目的　探讨TMD患者关节液中5 -HT和NPY的水平与关节源性疼痛的关系。方法　用ELISA法检测TMD患者的1 2 6侧关节和健康志愿者的32侧关节的关节液标本中的5 -HT ,用放射免疫测定法检测关节液标本中NPY的含量。1 2 6侧TMD患者的关节液标本分为疼痛组(n =90 )和无痛组(n =36 ) ;32名正常志愿者的32侧关节液设为对照组(n =32 )。结果　所有标本都检出5 -HT ,无痛组含量为1 78.75 0±6 4 .979ng/ml,疼痛组含量为2 2 3.5 2 2±82 .6 36ng/ml,对照组含量为1 36 .1 2 5±5 5 .0 75ng/ml。统计学检验:疼痛组与无痛组、疼痛组与对照组以及无痛组与对照组间比较均有显著性差异(P <0 .0 5 )。NPY检测结果:疼痛组1 33.5 0 0±4 6 .6 6 4 pg/ml,无痛组99.6 5 6±2 5 .873pg/ml,对照组83.2 86±2 8.397pg/ml。统计学检验:疼痛组与无痛组、疼痛组与对照组以及无痛组与对照组间比较均有显著性差异(P <0 .0 5 )。结论　关节液中5 -HT、NPY的水平,患者组显著高于正常人组,并且在患者中疼痛组又显著高于无痛组,表明患者关节内存在炎症状态,也表明关节疼痛与关节液中5 -HT、NPY的水平增高密切相关。     

Detection of, and anti-collagen antibody produced by, CD5-positive B cells in inflamed gingival tissues

This study was performed to investigate the frequency and distribution of CD5-positive (CD5+) B cells in inflamed gingival tissues using flow cytometric and immunohistochemical analyses. The ability of CD5+ B cells to produce anti-type I collagen antibody was also examined. CD5+ B cells expressed "low" fluorescence intensity in the peripheral blood of both healthy subjects and patients with adult periodontitis. However, in inflamed gingival tissues the intensity of this surface marker was high. The percentage of B cells bearing CD5 surface marker was statistically higher in gingiva than in peripheral blood obtained from both the patients and healthy subjects. These CD5+ B cells were observed in gingival subepithelial connective tissues from the bottom to the middle of the periodontal pocket. This area showed destruction of collagen fibers and dense cell infiltrations. Anti-collagen IgG antibody level in patients' gingival crevicular fluids (GCF) was higher than that in sera from healthy subjects, and slightly higher than in autologous sera. IgM anti-collagen antibody in GCF was lower than in autologous sera and in sera from healthy subjects. EBV-transformed CD5+ B cells produced considerably more IgM and IgG antibody to collagen than CD5- B cells. Therefore CD5+ B cells may contribute to the pathogenesis of inflamed gingival tissues.     

A functional interleukin-1 beta gene polymorphism is associated with chronic periodontitis in a sample of Brazilian individuals

BACKGROUND: Interleukin-1 beta (IL-1 beta) is a potent inflammatory mediator and an important polymorphism in the locus +3954 (C/T) of the human IL1 B gene has been shown to affect the levels of this cytokine. This functional polymorphism has been associated with the establishment of inflammatory diseases, including periodontal disease, in European, Asian and North American populations. OBJECTIVE: The aim of this study was to investigate the association between the IL1 B (+3954) gene polymorphism and the occurrence of different clinical forms of periodontitis in a sample of Brazilian individuals. METHODS: This study employed a cross-sectional design involving individuals from the State of Minas Gerais in the south-eastern region of Brazil. Genomic DNA was obtained from oral swabs of 129 individuals and amplified using the polymerase chain reaction (PCR) with specific primers flanking the locus +3954 of IL1 B. PCR products were submitted to restriction endonuclease digestion and analyzed by polyacrylamide gel electrophoresis, to distinguish alleles T and C of the IL1 B gene, allowing for the determination of the genotypes and detection of the polymorphism. RESULTS AND CONCLUSIONS: The chronic periodontitis group displayed a higher percentage of the T allele (28%) when compared to the aggressive periodontitis group (10.7%, chi(2)=5.24, p=0.02, OR=0.31, CI=0.11--0.88) and to control group (8.7%, chi(2)=7.11, p=0.007, OR=0.24, CI=0.08--0.73). Our data suggested that the polymorphism in the locus +3954 of IL1 B gene could be a risk factor for chronic periodontitis in a sample of Brazilian individuals.     

Association of vitamin D receptor gene polymorphisms in Chinese patients with generalized aggressive periodontitis

Background and Objective:  The clinical features suggest that genetic factors may have a strong influence on susceptibility to aggressive periodontitis. The aim of this study was to investigate the association of vitamin D receptor gene polymorphisms with generalized aggressive periodontitis in Chinese patients.
Material and Methods:  A restriction fragment length polymorphism (RFLP) for 10,438,141 C to T (rs1544410, Bsm I), 10,382,063 A to G (rs731236, Taq I), 10,382,143 C to A (rs7975232, Apa I) and 10,416,201 A to G (rs2228570, Fok I) of vitamin D receptor gene was analysed by polymerase chain reaction, followed by digestion with restriction enzymes and gel electrophoresis. The genotypes of 51 generalized aggressive periodontitis patients and 53 periodontally healthy control subjects were analysed. The genotypic and allelic frequencies of each polymorphism site for the patients and control subjects were compared.
Results:  The distribution of vitamin D receptor Fok I genotypes and alleles between the two groups was significantly different ( p =  0.043 and p  = 0.012, respectively). The F allele seemed to increase the susceptibility of aggressive periodontitis (odds ratio = 2.02, 95% confidence interval = 1.16–3.50) in Chinese patients. There was no significant difference in the genotype distribution or the allele frequencies of vitamin D receptor Bsm I, Apa I and Taq I between two groups.
Conclusion:  The study indicates that Fok I polymorphism of vitamin D receptor gene might be associated with generalized aggressive periodontitis in Chinese patients. In addition, the carriage of F allele increases the risk of developing generalized aggressive periodontitis.     

IL-1RN gene polymorphism is associated with peri-implantitis

Objectives: Interleukin (IL)‐1α, IL‐1β and their natural specific inhibitor IL‐1 receptor antagonist (IL‐1ra) play a key role in the regulation of the inflammatory response in periodontal tissues. Polymorphisms in the IL‐1 gene cluster have been associated with severe adult periodontitis. We aimed to investigate the IL‐1 gene cluster polymorphisms in patients with peri‐implantitis. Material and methods: The study included 120 North Caucasian individuals. A total of 71 patients (mean age 68 years, 76% smokers) demonstrating peri‐implantitis at one or more implants as evidenced by bleeding and/or pus on probing and bone loss amounting to >3 threads on Brånemark implants and 49 controls (mean age 66 years, 45% smokers) with clinical healthy mucosa and no bone loss around the implants were recruited for the study. The titanium implants, ad modum Brånemark, had been in function for at least 2 years. Mouthwash samples were collected and used for genotyping of the bi‐allelic polymorphisms IL‐1A^?889, IL‐1B⁺³⁹⁵³, IL‐1B^?511 and a variable number of tandem repeat IL‐1RN gene polymorphisms using PCR technique. Results: Significant differences were found in the carriage rate of allele 2 in the IL‐1RN gene between peri‐implantitis patients and controls (56.5% vs. 33.3%, respectively; odds ratios (OR) 2.6; 95% confidence interval (CI) 1.2–5.6; P=0.015). Logistic regression analysis taking smoking, gender and age into account confirmed the association between the IL‐1RN allele 2 carriers and peri‐implantitis (OR 3; 95% CI 1.2–7.6; P=0.02). Conclusions: Our results provide evidence that IL‐1RN gene polymorphism is associated with peri‐implantitis and may represent a risk factor for this disease.     

Influence of interleukin-1 gene polymorphism on periodontal regeneration in intrabony defects

The aim of this controlled retrospective study was to evaluate the influence of an IL-1 gene polymorphism on the clinical and radiographic healing outcomes of GTR therapy. The study included 47 adult periodontitis patients with 94 deep intrabony defects treated by GTR using different membrane materials. The following clinical parameters were recorded at baseline and 12 months after surgery: papillary bleeding index (PBI), gingival recession (REC), probing pocket depth (PPD), clinical attachment level (CAL), and the vertical relative attachment gain (V-rAG). Bone changes in the defect regions due to GTR therapy were quantitatively evaluated using digital subtraction radiography (DSR). Polymorphisms of the IL-1A gene at position - 889 and of the IL-1B gene at position + 3953 were analyzed by PCR. Statistical analysis was performed using the Mann-Whitney-U and the Wilcoxon-Signed-Rank tests (alpha = 0.05). The study comprised 19 IL-1 genotype positive (IL-1 +) patients and 28 IL-1 genotype negative (IL-1 -) patients. Twelve months after GTR therapy, both patient groups revealed statistically significant PPD reductions and CAL gain [median (25/75% percentiles)]: Delta PPD [IL-1 + : 4.0 (2.5/5.0) mm; IL-1-: 3.8 (3.0/4.9) mm], Delta CAL [IL-1 + : 3.5 (3.0/4.8) mm; IL-1 -: 3.0 (1, 2/4, 5) mm]. V-rAG amounted to 60.0 (47.7/78.6)% in IL-1 + patients and 53.1 (43.4/81.9)% in IL-1 - patients. Both patient groups showed significant bone density gain in 40% (IL-1 +) and 43.6% (IL-1 -) of the initial defect area due to GTR. Neither the clinical nor the radiographic healing parameters revealed any statistically significant differences in the GTR healing outcome between IL-1 + and IL-1 - patients. In conclusion, these 12-month findings indicate that the IL-1 gene polymorphism has no influence on the clinical and radiographic regeneration results following GTR therapy.     

Stretching stimulates fibulin-5 expression and controls microfibril bundles in human periodontal ligament cells

Background and Objective:  The elastic fiber system comprises oxytalan, elaunin and elastic fibers, differing in their relative microfibril and elastin contents. Human periodontal ligaments contain oxytalan fibers (pure microfibrils). Periodontal ligaments are continuously exposed to various functional forces, such as tooth movement and occlusal loading. We have reported that bundles of microfibrils coalesce in response to mechanical strain in cultured periodontal ligament fibroblasts, as assessed in terms of their positivity for fibrillin-1 (the major component of microfibrils). However, the mechanism of microfibril coalescence is unclear. We hypothesized that the fibrillin-1-binding molecule, fibulin-5, contributes to oxytalan fiber formation under mechanical strain.
Material and Methods:  We subjected periodontal ligament fibroblasts to stretching in order to examine the effects of fibulin-5 on the formation of oxytalan fibers in cell/matrix layers. We transfected periodontal ligament cells with small interference RNA for fibulin-5, then examined oxytalan fibers using immunofluorescence and electron microscopy.
Results:  Immunofluorescence showed that fibrillin-1-positive microfibrils coalesced as a result of stretching, compared with cells that were not subjected to stretching. Fibulin-5 colocalized on fibrillin-1-positive microfibrils. Stretching increased fibulin-5 gene expression and protein deposition. Immunofluorescence and immunogold electron microscopy analysis revealed that fibulin-5 suppression inhibited the coalescence of microfibrils under stretching conditions.
Conclusion:  These results suggest that fibulin-5 up-regulated in response to tension strain may control the formation of microfibril bundles in periodontal ligament.