Autoimmune Neutropenia as a Cause of Periodontal Disease in Preschool Children.

In autoimmune neutropenia, autoantibodies attack neutrophils resulting in their destruction or alteration of their function. Since neutrophils have important immunologic functions, aberrations in their homeostasis lead to increased susceptibility to diseases, such as periodontitis. Periodontitis as a manifestation of neutropenia can affect adults and children. In this paper, we describe the treatment of periodontal disease in a 2-year-old female with autoimmune neutropenia. The importance of an interdisciplinary approach, frequent recalls, and meticulous mechanical therapy in stabilizing her periodontal condition, despite ongoing systemic infections is emphasized.

Cytokine expression in peri-implant crevicular fluid in relation to bacterial presence.

AIM: The aim was to assess clinical inflammatory parameters, cytokine levels and bacterial counts in samples from implant crevicular fluid in cases with untreated peri-implantitis. MATERIAL AND METHODS: Several bacterial species known to up-regulate pro-inflammatory cytokines have been associated with peri-implantitis. The Luminex magnet bead technology was used to study cytokines in crevicular fluid. The checkerboard DNA-DNA hybridization method was used to study bacterial counts in samples from 41 implants (41 individuals). RESULTS: Profuse bleeding and suppuration was found in 25/41 (61.0%) of the implants. The reliability of duplicate cytokine processing was high. In the presence of profuse bleeding, higher pg/ml levels of IL-1ß (p = 0.02), IL-8 (p = 0.04), TNF-α (p = 0.03) and VEGF (p = 0.004) were found. Higher concentrations of IL-1ß were found in the presence of suppuration, and if Escherichia coli (p = 0.001) or Staphylococcus epidermidis (p = 0.05) could be detected. CONCLUSION: Profuse bleeding and/or suppuration in untreated peri-implantitis can be associated with higher concentrations of IL-1ß, IL-8, TNF-α and VEGF in peri-implant crevicular fluid. A higher concentration of IL-1ß in peri-implant crevicular fluid was found in samples that were positive for E. coli or S. epidermidis.

Tumor necrosis factor-α and oral inflammation in patients with Crohn disease.

BACKGROUND: Crohn disease (CD) is a chronic inflammatory bowel disease often accompanied by periodontal symptoms. Based on its function in immune response, tumor necrosis factor (TNF)-α and its genetic variants have been discussed as risk indicators in inflammatory processes. Therefore, the aim of the present study is to investigate the impact of TNF-α polymorphisms on periodontal parameters and inflammatory lesions of oral mucosa as a characteristic of CD. METHODS: A total of 142 patients with CD were included in the study. Oral soft tissue alterations and periodontal parameters were assessed. Genotypes, alleles, and haplotypes of TNF-α polymorphisms (rs1800629, cDNA-308G > A; and rs361525, cDNA-238G > A) were determined by polymerase chain reaction with sequence-specific primers (PCR-SSP). RESULTS: Patients with CD who exhibit more severe oral soft tissue alterations were significantly more often A allele carriers of rs361525 than G allele carriers (14.2% versus 2.2%; P <0.001). Furthermore, A allele carriers had a higher mean periodontal probing depth (P <0.05), mean clinical attachment level (P <0.05), and sites with bleeding on probing (not significant). Similar results were obtained when evaluating A allele-containing genotypes (AG + AA) and haplotypes (GA). In multivariate analyses considering age, sex, smoking, and medication as confounders, the A allele was proven to be an independent risk indicator for oral soft tissue alterations in patients with CD. No genotype-dependent influence of rs1800629 was observed. CONCLUSION: The TNF-α A allele of rs361525 represents a significant risk indicator for oral soft tissue alterations in patients with CD.

Relationship between chemokines and dendritic cells in human chronic periodontitis.

BACKGROUND: The purpose of this study was to evaluate the relationship between chemokines and dendritic cells (DCs) in human chronic periodontitis (CP). METHODS: Gingival samples were obtained from 23 individuals with CP, and six samples of normal mucosa (NM) overlapping the third molar were used to control for the chemokine levels. Periodontal examination was conducted. Immunohistochemistry was performed for Factor XIIIa(+) and cluster of differentiation (CD)1a(+) immature DCs and CD83(+) mature DCs. Levels of the CC chemokine ligand (CCL)2, CCL3, CCL5, CCL19, CCL20, and CXC chemokine ligand (CXCL)8 were measured in gingival tissues using enzyme-linked immunosorbent assay. Inflammatory infiltrate, DCs, chemokines, classification of human CP, and clinical parameters were correlated and compared. RESULTS: The expression of CCL2 and CCL20 was positively correlated with increased densities of CD1a(+) DCs. CCL3 and CXCL8 were positively related to the clinical attachment level. CCL3, CCL5, CCL19, and CXCL8 levels increased in the gingival samples of patients with CP compared with NM, whereas CCL20 levels increased in advanced CP compared with mild-moderate CP. CONCLUSIONS: More CD1a(+) immature DCs are related to CCL2 and CCL20. CCL3 and CXCL8 chemokines are related to a greater severity of human CP.

Effect of the menstrual cycle on inflammatory cytokines in the periodontium.

BACKGROUND AND OBJECTIVE: The effects of different levels of steroid hormones, as experienced during puberty, pregnancy and menopause, on the periodontium have been demonstrated, but changes in sex hormone levels during the menstrual cycle, and the influence of these changes on the periodontium, remain unresolved. The aim of this study was to investigate the effect of the menstrual cycle on the levels of interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) in gingival crevicular fluid and on periodontal clinical parameters, including the gingival bleeding index (GBI) and the modified gingival index (MGI), in periodontally healthy women. MATERIAL AND METHODS: Twenty-seven periodontally healthy women with a regular menstrual cycle were included in the study. Clinical parameters, including the GBI, the MGI and the simplified oral health index, were recorded during menstruation, ovulation and premenstruation phases (e.g. on days 1-2, 12-14 and 22-24, respectively) of the menstrual cycle. Gingival crevicular fluid and unstimulated saliva were collected, at each study phase, for assessment of IL-1ß, TNF-α, estrogen and progesterone. RESULTS: Both the GBI and the MGI increased significantly during the menstrual cycle, and were significantly higher during ovulation than during menstruation or premenstruation (p < 0.001). No significant change in the simplified oral health index was observed during the menstrual cycle ( p = 0.18). The levels of IL-1ß and TNF-α increased during the different phases of the menstrual cycle, but only the change in the TNF-α concentration was significant ( p < 0.05). CONCLUSION: This study indicated that changes occurring during the menstrual cycle influence the periodontium and induce inflammatory conditions.

Influence of periodontal disease, Porphyromonas gingivalis and cigarette smoking on systemic anti-citrullinated peptide antibody titres.

BACKGROUND: Anti-citrullinated protein antibody (ACPA) responses may precede clinical onset of rheumatoid arthritis. Porphyromonas gingivalis peptidylarginine deiminase can citrullinate proteins possibly inducing autoimmunity in susceptible individuals. AIM: To determine whether periodontitis, carriage of P. gingivalis, smoking and periodontal therapy influence ACPA titres. METHODS: Serum and plaque samples were collected from 39 periodontitis patients before and after non-surgical periodontal treatment, and from 36 healthy subjects. Carriage of P. gingivalis was determined by PCR of plaque DNA. ACPA was determined by anti-cyclic citrullinated peptide (CCP) enzyme-linked immunosorbent assay (ELISA). Anti-P. gingivalis titres were determined by ELISA. RESULTS: Untreated periodontitis patients had higher anti-CCP antibody titres than healthy controls [three patients (8%) greater than manufacturer suggested assay diagnostic threshold (5 Assay Units/AU) versus none (0%); mean ± SEM: 1.37 ± 0.23 versus 0.40 ± 0.10 AU, p < 0.0001]. Periodontitis patients who smoked demonstrated lower anti-P. gingivalis (15956 ± 4385 versus 2512 ± 1290 Units/ml, p < 0.05), but similar anti-CCP than non-smoking periodontitis patients (smokers: 1.31 ± 0.35; non-smokers: 1.41 ± 0.32 AU). Healthy smokers demonstrated elevated anti-CCP titres (0.75 ± 0.19 AU), at levels between healthy non-smokers (0.15 ± 0.05 AU) and non-smoker periodontitis patients. Six months after periodontal treatment, there were significant reductions in anti-CCP (non-smokers p < 0.05) and anti-P. gingivalis (all participants p < 0.01). CONCLUSION: In subjects with periodontitis, P. gingivalis infection may be responsible for inducing autoimmune responses that characterize rheumatoid arthritis.

Differential expression of mitogen activating protein kinases in periodontitis.

AIM: Following toll-like receptor (TLR) engagement, lipopolysaccharide (LPS) can stimulate the expression of pro-inflammatory cytokines thus activating the innate immune response. The production of inflammatory cytokines results, in part, from the activation of kinase-induced signalling cascades and transcriptional factors. Of the four distinct classes of mitogen-activated protein kinases (MAPK) described in mammals, p38, c-Jun N-terminal activated kinases (JNK1-3) and extracellular activated kinases (ERK1,2) are the best studied. Previous data have established that p38 MAPK signalling is required for inflammation and bone loss in periodontal disease pre-clinical animal models. MATERIALS & METHODS: In this study, we obtained healthy and diseased periodontal tissues along with clinical parameters and microbiological parameters. Excised fixed tissues were immunostained with total and phospho-specific antibodies against p38, JNK and ERK kinases. RESULTS: Intensity scoring from immunostained tissues was correlated with clinical periodontal parameters. Rank correlations with clinical indices were statistically significantly positive (p-value < 0.05) for total p38 (correlations ranging 0.49-0.68), phospho-p38 (range 0.44-0.56), and total ERK (range 0.52-0.59) levels, and correlations with JNK levels also supported association (range 0.42-0.59). Phospho-JNK and phospho-ERK showed no significant positive correlation with clinical parameters of disease. CONCLUSION: These data strongly implicate p38 MAPK as a major MAPK involved in human periodontal inflammation and severity.

Role of serum IL-23/IL-17 axis in the relationship between periodontitis and coronary heart disease.

To explore the role of the IL-23/IL-17 axis in the relationship between periodontitis and coronary heart disease (CHD), 97 subjects were recruited and divided into four groups: (1) CHD + periodontitis, (2) CHD, (3) periodontitus alone, and (4) healthy. The demographic characteristics and periodontal status of all subjects were recorded, and the serum levels of IL-23/IL-17 were detected by enzyme-linked immunoabsorbent assay. Results showed that the serum levels of IL-23/IL-17 in groups 1, 2, and 3 were higher compared with group 4. Group 1 manifested the highest level of serum IL-23/IL-17. A significant positive correlation between IL-23 and IL-17 levels was seen in the three patients groups; groups 1 and 3 also had significant positive correlations with probing depth and attachment loss. The results indicate that there may be an association between periodontitis and CHD, and the IL-23/IL-17 axis may play an important role in the pathologic process of both diseases.

Innate immune receptor expression in peri-implant tissues of patients with different susceptibility to periodontal diseases.

BACKGROUND: Although inflammation mediates the pathogenesis of periodontal diseases, the effects of innate immune responses on implant therapies have not been evaluated. Innate immune receptors, including toll-like-receptors (TLRs) and the receptor for advanced glycated end-products (RAGE), are upregulated within inflamed gingiva and are responsible for initiation of detrimental host responses. The aim of this study is to compare the expression of TLR2, TLR4, and RAGE in gingival tissues from participants susceptible to periodontitis and participants not susceptible to periodontitis before and after implant therapy. METHODS: Periodontally healthy participants received implant therapy for non-periodontal edentulism. Participants susceptible to periodontitis were diagnosed with chronic periodontitis prior to implant therapy. Gingival biopsies were collected from edentulous ridges before implant installation and from peri-implant mucosa 2 months after treatment. Histology, real-time PCR, and Western blot were used to evaluate levels of inflammatory infiltrate, TLR2, TLR4, and RAGE expression. RESULTS: Before implant therapy, elevated levels of RAGE were detected in gingival tissues from participants susceptible to periodontitis when compared to those from participants with healthy periodontiums, whereas no differences in the expression of TLR2 or TLR4 were detected. After implant therapy, there was an upregulation of RAGE and TLR4 levels that coincided with a downregulation of TLR2 levels in biopsies from participants susceptible to periodontitis. Levels of RAGE and TLR4 remained unchanged in biopsies from participants with healthy periodontiums, whereas TLR2 levels were significantly upregulated. Histologically, post-implant biopsies from participants susceptible to periodontitis displayed higher levels of inflammatory infiltrate. CONCLUSION: Elevated levels of inflammatory potential were found after implant therapy in participants susceptible to periodontitis.

Gingival fluid cytokine expression and subgingival bacterial counts during pregnancy and postpartum: a case series.

OBJECTIVES: The aim of this study was to assess gingival fluid (GCF) cytokine messenger RNA (mRNA) levels, subgingival bacteria, and clinical periodontal conditions during a normal pregnancy to postpartum. MATERIALS AND METHODS: Subgingival bacterial samples were analyzed with the checkerboard DNA-DNA hybridization method. GCF samples were assessed with real-time PCR including five proinflammatory cytokines and secretory leukocyte protease inhibitor. RESULTS: Nineteen pregnant women with a mean age of 32 years (S.D. ± 4 years, range 26-42) participated in the study. Full-mouth bleeding scores (BOP) decreased from an average of 41.2% (S.D. ± 18.6%) at the 12th week of pregnancy to 26.6% (S.D. ± 14.4%) at the 4-6 weeks postpartum (p < 0.001). Between week 12 and 4-6 weeks postpartum, the mean probing pocket depth changed from 2.4 mm (S.D. ± 0.4) to 2.3 mm (S.D. ± 0.3) (p = 0.34). Higher counts of Eubacterium saburreum, Parvimonas micra, Selenomonas noxia, and Staphylococcus aureus were found at week 12 of pregnancy than at the 4-6 weeks postpartum examinations (p < 0.001). During and after pregnancy, statistically significant correlations between BOP scores and bacterial counts were observed. BOP scores and GCF levels of selected cytokines were not related to each other and no differences in GCF levels of the cytokines were observed between samples from the 12th week of pregnancy to 4-6 weeks postpartum. Decreasing postpartum counts of Porphyromonas endodontalis and Pseudomonas aeruginosa were associated with decreasing levels of Il-8 and Il-1ß. CONCLUSIONS: BOP decreased after pregnancy without any active periodontal therapy. Associations between bacterial counts and cytokine levels varied greatly in pregnant women with gingivitis and a normal pregnancy outcome. Postpartum associations between GCF cytokines and bacterial counts were more consistent. CLINICAL RELEVANCE: Combined assessments of gingival fluid cytokines and subgingival bacteria may provide important information on host response.

Periodontal condition of patients with autoimmune diseases and the effect of anti-tumor necrosis factor-α therapy.

BACKGROUND: The aim of this study is to evaluate the effect of autoimmune diseases (AIs), as well as anti-tumor necrosis factor-α (TNF-α) therapy on the clinical and immunologic parameters of the periodontium. METHODS: Thirty-six AI patients (12 rheumatoid arthritis [RA], 12 psoriatic arthritis, and 12 systemic sclerosis) were recruited together with 12 healthy (H) and 10 RA patients receiving anti-TNF-α therapy (RA+). Periodontal indices including plaque index, gingival index (GI), probing depth (PD), and bleeding on probing (BOP) were measured, and gingival crevicular fluid (GCF) was collected from five deepest pockets using papers strips. The TNF-α level was analyzed using enzyme-linked immunosorbent assay. Analysis of variance test was used for statistical comparison between groups, whereas Pearson linear correlation coefficient test was used to examine the association between TNF-α and periodontal status indices. RESULTS: The three AI subgroups were very similar in clinical and immunologic parameters. GI was greater in the AI patients compared to the H and RA+ groups (1.91 ± 0.54, 1.21 ± 0.67, and 1.45 ± 0.30, respectively, P = 0.0005). AI patients exhibited significantly more BOP than H and RA+ (46.45% ± 17.08%, 30.08% ± 16.86%, and 21.13% ± 9.51%, respectively, P = 0.0002). PD in H and RA+ groups were lower than in the AI (3.47 ± 0.33, 3.22 ± 0.41, and 3.91 ± 0.49 mm, P = 0.0001). Number of sites with PD >4 mm was higher in AI patients compared to H and RA+ (42.44 ± 17.5 versus 24.33 ± 15.62 versus 33.3 ± 6.6, P = 0.0002). GCF TNF-α was higher among the AI patients (1.67 ± 0.58 ng/site) compared to 1.07 ± 0.33 ng/site for the H group and 0.97 ± 0.52 ng/site for the RA+ group (P = 0.0002). A significant positive correlation was found between PD and TNF-α levels in the GCF (r = 0.4672, P = 0.0002), BOP (r = 0.7491, P = 0.0001), and GI (r = 0.5420, P = 0.0001). CONCLUSIONS: Patients with AI diseases have higher periodontal indices and higher TNF-α levels in GCF than H controls. Anti-TNF-α treatment appears to reverse this phenomenon.

Connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: microbiological and immunological results.

OBJECTIVES: It was previously reported the clinical results of placing subgingival resin-modified glass ionomer restoration for treatment of gingival recession associated with non-carious cervical lesions. The aim of this study was to evaluate the influence of this treatment on the subgingival biofilm and gingival crevicular fluid (GCF) inflammatory markers. MATERIALS AND METHODS: Thirty-four patients presenting the combined defect were selected. The defects were treated with either connective tissue graft plus modified glass ionomer restoration (CTG+R) or with connective tissue graft only (CTG). Evaluation included bleeding on probing and probing depth, 5 different bacteria targets in the subgingival plaque assessed at baseline, 45, and 180 days post treatments, and 9 inflammatory mediators were also assessed in the GCF. RESULTS: The levels of each target bacterium were similar during the entire period of evaluation (p > 0.05), both within and between groups. The highest levels among the studied species were observed for the bacterium associated with periodontal health. Additionally, the levels of all cyto/chemokines analyzed were not statistically different between groups (p > 0.05). CONCLUSION: Within the limits of the present study, it can be concluded that the presence of subgingival restoration may not interfere with the subgingival microflora and with GCF inflammatory markers analyzed. CLINICAL RELEVANCE: This approach usually leads to the placement of a subgingival restoration. There is a lack of information about the microbiological and immunological effects of this procedure. The results suggest that this combined approach may be considered as a treatment option for the lesion included in this study.

Comparison of CCL28, interleukin-8, interleukin-1ß and tumor necrosis factor-alpha in subjects with gingivitis, chronic periodontitis and generalized aggressive periodontitis.

BACKGROUND AND OBJECTIVE: Cytokines produced by various cells are strong local mediators of inflammation. Mucosa-associated epithelial chemokine (CCL28), interleukin-8 (IL-8), interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) are major cytokines that play important roles in the periodontal inflammatory process. In this study we aimed to compare the levels of CCL28, IL-8, IL-1ß and TNF-α in the gingival crevicular fluid of both periodontally healthy subjects and in subjects diagnosed with gingivitis, chronic periodontitis and generalized aggressive periodontitis. MATERIAL AND METHODS: A total of 84 subjects participated in the study: 21 subjects had gingivitis, 21 subjects had chronic periodontitis, 21 subjects had generalized aggressive periodontitis and 21 were periodontally healthy. The levels of CCL28, IL-8, IL-1ß and TNF-α were analyzed using enzyme-linked immune sorbent assay (ELISA). RESULTS: The total levels of CCL28 and IL-8 in the gingival crevicular fluid of the generalized aggressive periodontitis group (324.74 ± 42.62 pg/30 s, 487.62 ± 49.21 pg/30 s) were significantly higher than those of the chronic periodontitis group (268.81 ± 28.64 pg/30 s, 423.65 ± 35.24 pg/30 s), the gingivitis group (146.35 ± 17.46 pg/30 s, 310.24 ± 48.20 pg/30 s) and the periodontally healthy group (92.46 ± 22.04 pg/30 s, 148.41 ± 24.64 pg/30 s). Similarly, the total levels of IL-1ß and TNF-α in the generalized aggressive periodontitis group (110.23 ± 9.20 pg/30 s, 1284.46 ± 86.32 pg/30 s) were significantly higher than those in the chronic periodontitis group (423.65 ± 35.24 pg/30 s, 82.64 ± 9.12 pg/30 s), the gingivitis group (52.10 ± 7.15 pg/30 s, 824.24 ± 44.68 pg/30 s) and the periodontally healthy group (36.44 ± 8.86 pg/30 s, 628.26 ± 34.61 pg/30 s). CONCLUSION: CCL28, IL-8, IL-1ß and TNF-α may play key roles in the host response to inflammation in periodontal diseases. As the severity of periodontal diseases increases, destruction of periodontal tissues also increases. Inflammation is one among many factors that trigger periodontal tissue destruction. Identification of the mediators that influence the development and progression of inflammation in periodontal diseases may be very important in understanding the prognoses of periodontal diseases.

Expression of peptidylarginine deiminase-2 and -4, citrullinated proteins and anti-citrullinated protein antibodies in human gingiva.

BACKGROUND AND OBJECTIVE: The presence of citrullinated proteins, and peptidylarginine deiminase types -2 (PAD-2) and -4 (PAD-4) in periodontal tissues, determine the presence of anti-cyclic citrullinated protein antibodies (anti-CCP) in gingival crevicular fluid (GCF) and compare the expression of these proteins between inflamed and non-inflamed sites. MATERIAL AND METHODS: Tissue sections were stained using antibodies against citrullinated proteins, PAD-2 and PAD-4. RT-PCR was performed to investigate PAD-2 and PAD-4 mRNA in inflamed and non-inflamed gingival tissues. Anti-CCP antibodies in gingival crevicular fluid were detected by ELISA. RESULTS: Citrullinated proteins, PAD-2 and PAD-4 were detected in gingiva. There was a correlation between inflammation and expression of these proteins. mRNAs for PAD-2 and PAD-4 were detected in both inflamed and non-inflamed gingival tissues. Antibodies to CCP were found mostly in the GCF of individuals with periodontitis. CONCLUSION: PAD-2 and PAD-4 (protein and mRNA) as well as citrullinated proteins are present in inflamed gingiva, and anti-CCP antibodies can be detected in the GCF of some patients. Tissue expression of citrullinated proteins and PAD increased with the severity of inflammation. The presence of anti-CCP antibodies in GCF was almost exclusive to a subset of patients with periodontitis. Increased expression of these proteins in inflamed gingiva lends support to the notion that periodontal inflammation contributes to the inflammatory burden in a similar way to rheumatoid arthritis.

Effects of periodontal therapy on phagocytic activity of peripheral blood neutrophils - evidence for an extrinsic cellular defect.

PURPOSE: To compare phagocytic activity of peripheral blood neutrophils from subjects with and without periodontal disease and evaluate the effects of periodontal therapy in individuals with similar levels of resolution of inflammation at the end of treatment. MATERIALS AND METHODS: To compare the phagocytic activity of neutrophils, peripheral blood was collected from 27 control subjects with a healthy periodontium and 28 periodontitis subjects before and after treatment. The phagocytosis of killed Saccharomyces cerevisiae, pre-sensitised or non-sensitised with fresh serum from the donor, was quantified and a phagocytic index was calculated as the mean number of yeast cells phagocytised by the percentage of neutrophils involved in phagocytosis. RESULTS: Prior to periodontal treatment, subjects with periodontitis exhibited significantly lower neutrophil phagocytic activity than control subjects with a healthy periodontium. Periodontal treatment significantly improved in clinical periodontal status and resulted in significantly increased phagocytosis of both pre-sensitised (from 113.0 pre- to 157.0 post-treatment, P = 0.02) and non-sensitised S. cerevisiae (from 1.5 pre- to 3.5 post-treatment, P = 0.001), to levels observed in control subjects. CONCLUSIONS: The phagocytic activity of peripheral blood neutrophils from subjects with periodontal disease was lower than that of healthy controls. Subjects who underwent non-surgical periodontal treatment and strict supportive therapy for 6 months showed improved phagocytic activity in peripheral blood neutrophils. The phagocytic index values from subjects with periodontal disease after treatment achieved those found in the control group.

The importance of genetic variants in TNFα for periodontal disease in a cohort of coronary patients.

AIM: The aim of this analysis was to evaluate the importance of genetic variants of TNFα for the severity of periodontal disease and periodontal risk factors with respect to periodontal risk factors in a cohort of coronary patients. SUBJECTS: A total of 942 consecutive patients with angiographic proven coronary heart disease were prospectively included in the study entitled "Periodontitis and Its Microbiological Agents as Prognostic Factors in Patients With Coronary Heart Disease" (ClinicalTrials.gov identifier:NCT01045070). METHODS: After including of patients, an extensive periodontal examination also involving PCR-sampling for 11 periodontal bacteria was performed. In this subanalysis, single nucleotide polymorphisms (SNPs) c.-308G>A, c.-238G>A and haplotypes for TNFα were analysed by CTS-PCR-SSP Tray kit (Heidelberg, Germany). RESULTS: The AG+AA genotype of SNP c.-238G>A of TNFα gene was associated with the amount of clinical attachment loss in patients with coronary heart disease in multivariate regression analysis. Moreover, Prevotella intermedia occurred more frequently in carriers who were positive for the AG+AA genotype and A-allele of SNP c.-308G>A in bivariate and multivariate analyses. Furthermore, only in bivariate analyses significant associations of genetic variants of TNFα with intensified bleeding on probing and with higher plasma level of interleukin 6 could be shown. CONCLUSIONS: Genetic variants of TNFα gene, namely c.-308G>A and c.-238G>A, are associated with periodontal conditions in patients with coronary heart disease.

Bacterial and inflammatory behavior of implants in the early healing phase of chronic periodontitis.

OBJECTIVE: To assess the pattern of early bacterial colonization at implants and teeth in patients with a history of chronic periodontitis compared with a group of healthy subjects. Furthermore, the presence of host-derived markers at teeth and implants in the two subject groups was determined. METHOD AND MATERIALS: Subgingival and submucosal plaque and gingival crevicular fluid samples from 37 nonsubmerged healing dental implants and the deepest tooth sites per quadrant were analyzed 2 to 5 months after implant insertion. The presence of periodontal pathogens was assessed by means of real-time polymerase chain reaction. Further, the levels of interleukin (IL)-1Β, IL-8, and IL-10; secretory leukocyte protease inhibitor; and the neutrophil elastase activity were determined. RESULTS: Eleven patients with chronic periodontitis and 13 subjects without periodontitis were recruited for this study. Bacterial species associated with periodontitis were detectable at both the teeth and implants. The presence was always higher in the chronic periodontitis group; the difference was significant for Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans at both the implants and teeth. The levels of IL-1Β were higher at teeth than at implants; in contrast, more IL-10 was measured at the implants. CONCLUSION: The present results indicate that (1) dental implants inserted in periodontally compromised patients are colonized with periodontal pathogens within the first weeks of healing; (2) inflammatory markers (IL-1Β) are present in higher levels at teeth as compared with implants, whereas at implants, anti-inflammatory cytokines (IL-10) might play the important role; and (3) the importance of periodontal treatment prior to implant insertion to reduce bacterial load and inflammation should be emphasized.

Inflammatory biomarkers in saliva: assessing the strength of association of diabetes mellitus and periodontal status with the oral inflammatory burden.

AIM: To determine the strength of association of type 2 diabetes mellitus and periodontal disease with the oral inflammatory burden, as assessed by markers of inflammation in saliva. MATERIAL AND METHODS: Unstimulated saliva samples were collected from 192 subjects with or without type 2 diabetes. ß-glucuronidase (ßG) was measured via a fluorometric array and interlukin-1ß (IL-1ß) via enzyme-linked immunosorbent assay. The concentration of both mediators was evaluated in relationship to clinical parameters, severity of periodontal disease and diabetes status. RESULTS: Regression analysis demonstrated that diabetes and periodontal disease was independently and positively correlated with increased concentration of ßG in saliva (p < 0.001). Moreover, the relative association of periodontal disease with the level of ßG in saliva was greater than the strength of association of the diabetic status. IL-1ß concentration in saliva was primarily associated with the severity of periodontal disease (p < 0.01), but not the presence of diabetes (p = 0.50). CONCLUSIONS: This study examined the nature of the inflammatory response in the oral cavity as assessed by inflammatory markers in saliva. Both periodontal disease and diabetes mellitus were independently associated with the oral inflammatory burden, in which the effect of periodontal disease was more pronounced.

Interleukin-33 levels in gingival crevicular fluid, saliva, or plasma do not differentiate chronic periodontitis.

BACKGROUND: This study investigates whether gingival crevicular fluid (GCF), saliva, and plasma levels of interleukin-33 (IL-33) can differentiate individuals with chronic periodontitis from individuals with healthy periodontium. METHODS: GCF, whole saliva, and plasma samples together with full-mouth clinical periodontal recordings were obtained from 32 otherwise healthy, non-smoker chronic periodontitis individuals and 25 systemically and periodontally healthy, non-smoker individuals. IL-33 levels in the biofluid samples were determined by enzyme-linked immunosorbent assay. Data were tested statistically by Mann-Whitney U test. RESULTS: The GCF concentrations of IL-33 were significantly lower in chronic periodontitis individuals than in healthy individuals (P <0.0001), whereas the total amounts in GCF samples were similar (P >0.05). The salivary and plasma contrations of IL-33 were indifferent in the two study groups (P >0.05). CONCLUSIONS: According to the present findings, the GCF, saliva or plasma levels of IL-33 could not differentiate chronic periodontitis individuals and periodontally healthy individuals. Larger-scale intervention studies may better clarify this issue.

Proinflammatory cytokine levels in hyperlipidemic patients with periodontitis after periodontal treatment.

OBJECTIVE: The aim of this study was to evaluate the effects of periodontal treatment on serum and gingival crevicular fluid (GCF) proinflammatory cytokine levels in hyperlipidemic patients with periodontitis. MATERIALS AND METHODS: Fifty-two patients with hyperlipidemia and periodontitis and 28 systemically healthy controls with periodontitis (C) were included in the study. Hyperlipidemic groups were divided into two groups as suggested diet (HD) and prescribed statin (HS). The clinical periodontal parameters, fasting venous blood, and GCF samples were obtained, and serum tumor necrosis factor-alpha (TNF-α), interleukin (IL) 1-beta, and IL-6 levels were evaluated at baseline and at 3 months follow-up (3MFU) after the completion of the non-surgical periodontal treatment that included scaling and root planning. RESULTS: Percentage of bleeding on probing was significantly higher in the HS group than both the HD and C groups. In the HD and HS groups, there were significant decreases in serum IL-6 and GCF TNF-α levels between the 3MFU and baseline. A significant decrease was also found in GCF IL-6 at the end of the study period in the HS group. CONCLUSION: The combination of the periodontal therapy and antilipemic treatment may provide beneficial effects on the metabolic and inflammatory control of hyperlipidemia.