Search

An ability to predict the response to conventional non-surgical treatment of a periodontal site would be advantageous. However, biomarkers or tests devised to achieve this have lacked sensitivity. The aim of this study is to assess the ability of a novel combination of biomarkers to predict treatment outcome of patients with chronic periodontitis.Gingival crevicular fluid (GCF) and subgingival plaque were collected from 77 patients at three representative sites, one healthy (probing depth [PD] ≤3 mm) and two diseased (PD ≥6 mm), at baseline and at 3 and 6 months after treatment. Patients received standard non-surgical periodontal treatment at each time point as appropriate. The outcome measure was improvement in probing depth of ≥2 mm. Concentrations of active enzymes (matrix metalloproteinase [MMP]-8, elastase, and sialidase) in GCF and subgingival plaque levels of Porphyromonas gingivalis, Tannerella forsythia, and Fusobacterium nucleatum were analyzed for prediction of the outcome measure.Using threshold values of MMP-8 (94 ng/μL), elastase (33 ng/μL), sialidase (23 ng/μL), and levels of P. gingivalis (0.23%) and T. forsythia (0.35%), receiver operating characteristic curves analysis demonstrated that these biomarkers at baseline could differentiate healthy from diseased sites (sensitivity and specificity ≥77%). Furthermore, logistic regression showed that this combination of these biomarkers at baseline provided accurate predictions of treatment outcome (≥92%).The "fingerprint" of GCF enzymes and bacteria described here offers a way to predict the outcome of non-surgical periodontal treatment on a site-specific basis.

Objective To study the severity of bone loss in a Medieval UK population, and compare this with other ancient UK populations from different time periods. Design Skulls from a burial site in York (XI–XV century) were investigated. Skulls with a minimum of 17 teeth were included and were divided from childhood into five age groups. Direct measurements from the cement enamel junction (CEJ) to the alveolar crest (AC) were undertaken at six points around each tooth with a UNC 15 probe. The mean bone loss was calculated for each tooth type. Results Seventy five skulls were included in the study (12–60 years of age). Bone loss was found to increase with age, but stabilized in older individuals (>45 years). The mean CEJ-AC distance varied from 2.1 mm in the youngest group to a maximum of 4.1 mm in 36–45 year olds. Results were compared with a Roman–British population (Whittaker et al., 1982) where comparable values for mean bone loss were 2.2 mm and 5.4 mm respectively, and a population of XVIII century Londoners (Whittaker et al., 1998) where bone loss of 1.1 mm and 4.0 mm was reported for the youngest and oldest age groups respectively. Conclusion Measuring the CEJ–AC distance in dried skulls from ancient populations may be used as a proxy for the levels of periodontal disease irrespective of tooth wear. The findings from the current study suggest that the severity of periodontitis as determined by measurements of alveolar bone loss on dried skulls from this ancient population, seems to have declined in the United Kingdom from the III–V century to XVIII century. This may be due to changes in environmental factors including living conditions and diet, together with individual characteristics including systemic illness and genetic make up.

Aim To explore whether subjects harbouring A. actinomycetemcomitans, P. gingivalis or T. forsythia at baseline showed increased clinical benefits with the adjunctive use of systemic amoxicillin and metronidazole (AMX-MET) during non-surgical treatment of generalized aggressive periodontitis (GAgP). Material and Methods Forty one subjects were included in this 6-month randomized placebo-controlled clinical trial using a 7-day course of systemic AMX-MET or placebo as adjuncts to non-surgical periodontal therapy. Clinical and microbiological parameters were collected at baseline, 2 and 6 months after treatment. Microbiological cultures were processed for pooled subgingival samples and identities of isolates were determined by PCR for A. actinomycetemcomitans, P. gingivalis and T. forsythia Results At 6 months, the test treatment resulted in significant additional improvements in the primary outcome variable compared to placebo, and the effect of the adjunctive antimicrobials was not modified by the baseline microbiological status in the primary analysis. However, secondary exploratory subgroup analyses showed improved clinical outcomes in subjects harbouring A. actinomycetemcomitans at baseline compared to subjects who did not harbour this pathogen. Conclusions All subjects benefited from the tested adjunctive antimicrobial regimen, although subjects who harboured A. actinomycetemcomitans at baseline may show greater clinical benefits. Larger appropriately powered studies are needed to confirm whether adjunctive AMX-MET is more beneficial for GAgP patients who harbour A. actinomycetemcomitans, along with other key periodontal pathogens.

Aim To determine whether combinations of enzymes in gingival crevicular fluid (GCF) can act as improved biomarkers compared with single enzymes for predicting the outcome of treatment and also for diagnosing the clinical status of sites. Methods Thirty subjects with chronic periodontitis were recruited to a 12-month longitudinal pilot study. GCF samples from three representative sites: healthy (≤3 mm), deep non-bleeding (NB) (≥6 mm) and deep bleeding (DB) (≥6 mm) sites and clinical data were collected at baseline, 3 months, 6 months and 12 months following periodontal treatment. Active enzyme levels (MMP-8, cathepsin G, elastase, trypsin-like activity and sialidase) in GCF samples were assessed. The enzyme profiles and clinical data of each site were analysed for correlation and logistic regression was performed to find the predictive value of the active enzyme levels regarding the outcome of treatment. Results Twenty-two individuals completed the study. All active enzyme levels were significantly higher in diseased sites than healthy sites. Logistic regression showed that the combination of MMP8, elastase and sialidase provided accurate predictions of treatment outcome (88% for NB and 86% for DB), which was significantly better than each enzyme alone (61%). Conclusion This pilot has suggested that combined active enzyme profiling could provide significant prediction of outcome of treatment.

Griffiths GS, Ayob R, Guerrero A, Nibali L, Suvan J, Moles DR, Tonetti MS. Amoxicillin and metronidazole as an adjunctive treatment in generalized aggressive periodontitis at initial therapy or re-treatment: a randomized controlled clinical trial. J Clin Periodontol 2011; 38: 43–49. doi: 10.1111/j.1600-051X.2010.01632.x Abstract Background: Previously, we showed that systemic metronidazole and amoxicillin significantly improved the outcomes of non-surgical debridement in generalized aggressive periodontitis patients. This study aimed to observe whether re-treatment with adjunctive antimicrobials would give the placebo group benefits comparable with the test group. Methods: Thirty-eight of 41 subjects, from the initial 6-month trial, completed the second phase, re-treatment of sites with remaining pockets 5 mm. Subjects on placebo in phase one, received adjunctive antibiotics for 7 days. Clinical parameters were collected at 2 months posttreatment (8 months from baseline). Results: Patients who received antibiotics at initial therapy, showed statistically significant improvement in pocket depth reduction and in the % of sites improving above clinically relevant thresholds, compared with patients who received antibiotics at re-treatment. In deep pockets (7 mm), the mean difference was 0.9 mm (p=0.003) and in moderate pockets (4–6 mm) it was 0.4 mm (p=0.036). For pockets converting from 5 to 4 mm, this was 83% compared with 67% (p=0.041) and pockets converting from 4 to 3 mm was 63% compared with 49% (p=0.297). Conclusions: At 8 months, patients who had antibiotics at initial therapy showed statistically significant benefits compared with those who had antibiotics at re-treatment.

Aim: The aim of this analysis was to investigate the relationship between a vitamin D receptor (VDR) polymorphism and the diagnosis and progression of periodontitis. Material and Methods: Data were derived from two different studies, including 231 subjects with healthy periodontium, 224 aggressive periodontitis and 79 chronic periodontitis (CP) patients in a case-control investigation. Sixty-one of these CP patients also took part in an observational study with a 1-year follow-up, in which progression of periodontitis was determined at the subject level. All 534 subjects provided a blood sample from which genomic DNA was extracted to study VDR - 1056 TaqI polymorphism. Results: The interaction between smoking and VDR polymorphism was associated with the diagnosis of periodontitis in Caucasians [p = 0.001, odds ratio (OR) = 1.33, 95% confidence intervals (CI) = 1.12-1.57] and all subjects (p = 0.033, OR = 1.60, 95% CI = 1.04-2.48). In the longitudinal study, subjects were divided into two clusters at 1 year according to the median number of progressing sites (Acumulative attachment loss >2mm). Logistic regression analysis revealed that the interaction between VDR Taq-I polymorphism and smoking showed limited evidence of association with the "severe progression" cluster (p = 0.033, OR = 15.24, 95% CI= 1.24-187.42). Conclusions: Vitamin D receptor Taq-I TT polymorphism was moderately associated with both the presence and the progression of periodontitis in smokers, while no association was detected in non-smoking individuals. VDR genetic factors may interact with smoking in the pathogenesis of periodontitis.

This study reports the radiographic analysis of a split-mouth, single-blinded, randomised controlled clinical trial which was designed to compare the efficacy of simplified papilla preservation flap (SPPF) with or without guided tissue regeneration (GTR) in patients with aggressive periodontitis (AgP). Eighteen AgP patients who had similar bilateral intrabony defects were treated. In all patients, the defects presented with radiographic evidence of an intrabony defect ≥3 and ≥5 mm of periodontal pocket depths (PPD). The surgical procedures included access for root instrumentation using SPPF alone (control) or, after debridement, a placement of resorbable GTR membrane (test). The standardised radiographic assessments were carried out at pre-surgical baseline and at 6 and 12 month post-surgery. Radiographic linear measurements and subtraction radiography were used as the method of analysis. Both treatments showed significant improvements in linear radiographic bone fill and defect resolution at 6 and 12 months, compared to baseline. The 12-month subtraction radiography at the GTR sites showed a significant improvement compared to the 6-month outcomes. Both therapies were effective in the treatment of intrabony defects in AgP patients although no significant differences between them could be demonstrated. The finding that the bone fill and resolution of the defect at the GTR sites were significantly higher at 12 months than at 6 months after treatment indicates that bone regeneration is still an ongoing process at 6 months post-surgery. Radiographic assessment of periodontal regeneration should be carried out at 12 months post-surgery in order to evaluate the complete healing of the bony defect.

The aim of this study was to compare clinical outcomes between guided tissue regeneration (GTR) and access flap (AF) surgery in patients with aggressive periodontitis (AgP). Eighteen AgP patients with similar bilateral intrabony defects were treated in this split-mouth, single-blinded, randomised, controlled clinical trial. All patients presented with ≥3 mm intrabony defects and ≥5 mm periodontal pocket depths (PPD). In each patient, one defect was treated with a polyglycolide membrane according to the GTR principle, whereas the contralateral side was treated with AF. For both sides, a simplified papilla preservation flap was used. At baseline, 6 and 12 months post-surgery, the clinical attachment levels (CAL) and PPD were evaluated. At 6 and 12 months, at the GTR sites, the mean [95 % CI] CAL gain was 1.7 mm [1.1, 2.3] and 1.6 mm [0.9, 2.1], respectively, while the mean [95 % CI] PPD reduction was 2.3 mm [1.9, 2.8] and 2.4 mm [1.9, 2.8], respectively. Similar CAL (1.6 mm [1.0, 2.2] and 2.1 mm [1.4, 2.7]) and PPD (2.0 mm [1.5, 2.4] and 2.5 mm [2.0, 3.0]) outcomes were observed at the control sites at 6 and 12 months, respectively. Notably, at the GTR-treated sites, 13 subjects presented with various degrees of membrane exposure. Both therapies were effective in the treatment of intrabony defects in AgP patients, and no statistically significant differences between them could be demonstrated, possibly as a result of the differing degrees of membrane exposure at the GTR sites. Both periodontal regeneration and conventional periodontal surgery are effective treatments for AgP patients.

Background: The objective of this study was to assess the adjunctive clinical effect of the administration of systemic amoxicillin and metronidazole in the non-surgical treatment of generalized aggressive periodontitis (GAP).Methods: Forty-one systemically healthy subjects with GAP were included in this 6-month double-blind, placebo-controlled, randomized clinical trial. Patients received a course of full-mouth non-surgical periodontal treatment delivered over a 24 h period using machine-driven and hand instruments. Test subjects received an adjunctive course of systemic antibiotic consisting of 500 mg amoxicillin and 500 mg metronidazole three times a day for 7 days. Clinical parameters were collected at baseline, and at 2 and 6 months post-treatment.Results: In both the test and the placebo groups, all clinical parameters improved at 2 and 6 months. In deep pockets ( >= 7 mm), the test treatment resulted in an additional 1.4 mm (95% confidence interval 0.8, 2.0 mm) in full-mouth probing pocket depth (PPD) reduction and 1 mm (0.7, 1.3 mm) of life cumulative attachment loss (LCAL) gain at 6 months. In moderate pockets (4-6 mm), the adjunctive benefit was smaller in magnitude: PPD reduction was 0.4 mm (0.1, 0.7 mm) and LCAL gain was 0.5 mm (0.2, 0.8 mm). In addition, the 6-month data showed LCAL gains >= 2 mm at 25% of sites in test patients compared with 16% in placebo (p = 0.028). Similarly, PPD reductions of 2 mm or more were observed in 30% of sites in test and 21% of sites in placebo patients. Seventy-four percent of pockets with PPD >= 5 mm at baseline were 4 mm or shallower at 6 months in the test group. This compared with 54% in the placebo group (p = 0.008). Disease progression at 6 months was observed at 1.5% of test and 3.3% of sites in test and placebo, respectively (p = 0.072).Conclusions: These data indicate that a 7-day adjunctive course of systemic metronidazole and amoxicillin significantly improved the short-term clinical outcomes of full-mouth non-surgical periodontal debridement in subjects with GAP.

Objectives . Previous periodontal literature has shown that there is a strong relationship between treatment effects, such as guided tissue regeneration (GTR), and baseline disease severity. However, relating change to baseline values using correlation or regression is methodologically flawed due to mathematical coupling, where the statistical procedure of testing the null hypothesis—that the coefficient of correlation or slope of regression is equal to zero—becomes erroneous. The aim of this study is to investigate if baseline disease severity is genuinely associated with the treatment outcome of intrabony defects using GTR after adjustment for mathematical coupling. In particular, we seek to demonstrate the potential effect that mathematical coupling has in distorting the results from the statistical analyses of trials of dental treatment, using data from the periodontal literature on GTR. The erroneous results arising from the use of simple correlation and regression techniques to analyse this association will be demonstrated, also the methodological flaw where the statistical procedure tests the null hypothesis—that the coefficient of correlation or the slope of regression is equal to zero. Methods . Three main periodontal journals were electronically and manually searched to extract the data for the clinical outcomes of pocket probing depth (PPD) and lifetime cumulative attachment loss (LCAL) in the studies using GTR. The relationship between clinical outcomes and baseline measurements were reanalysed using Oldham's method and the variance ratio test. Results . The results of these analyses were compared with those from the papers where the authors used the standard approach of correlation or regression. This shows that mathematical coupling caused spurious correlations between baseline disease severity and treatment effect. Ten out of 12 studies for PPD and nine out of 14 for LCAL initially claimed a significant positive relationship; after using either of the more appropriate statistical methods of adjustment, only three correlations in each group of studies remained significant. Conclusions : Previous evidence suggesting an association between baseline disease severity and treatment effect for GTR is challenged and therefore needs to be critically reviewed. All future clinical research should avoid using mathematically coupled data in correlation or regression analysis. In seeking to examine the bivariate association between baseline and subsequent change, Oldham's method is recommended.

Animal studies report photodynamic therapy (PDT) to improve healing of excisional wounds but the mechanism is uncertain and equivalent human studies are lacking. To explore the impact of methyl aminolaevulinate (MAL)-PDT on clinical and microscopic parameters of human cutaneous excisional wound healing, examining for potential modulation through production of transforming growth factor (TGF)-β isoforms. In healthy older men (60-77 years; n=27), a 4 mm punch biopsy wound was created in skin of the upper inner arm and treated with MAL-PDT three times over five days. An identical control wound to the contralateral arm was untreated and both wounds left to heal by secondary intention. Wounds were re-excised at time points examining the inflammatory phase (7 days, n=10), matrix remodelling (3 weeks, n=8) and cosmetic outcome/dermal structure (9 months, n=9). Production of TGF-β1, TGF-β3 and matrix metalloproteinases (MMPs), key mediators of matrix deposition and remodelling, was assessed by immunohistochemistry alongside microscopic measurement of wound size/area and clinical assessment of wound appearance. MAL-PDT delayed re-epithelialisation at 7 days, associated with increased inflammation. However, 3 weeks post-wounding, treated wounds were smaller with higher production of MMP-1 (P=0.01), MMP-9 (P=0.04) and the anti-scarring cytokine, TGF-β3 (P=0.03). TGF-β1 was lower than control at 7 days and higher at 3 weeks (both P=0.03). At 9 months MAL-PDT treated wounds showed greater, more ordered deposition of collagen I, collagen III and elastin (all P

Aim: The aims of this study were to monitor the prevalence and progression of lifetime cumulative attachment loss (LCAL) in a group of young British male military recruits over a 3-year period, and to determine the relationship between signs of LCAL and selected periodontal variables.Methods: 100 subjects, aged 16-20 years (mean 17 years) at baseline. were examined at 0 (baseline), 12 and 30 months. LCAL, probing depth, plaque, bleeding on probing, gingival colour and supra- and subgingival calculus were assessed on the mesio-buccal, disto-buccal, mesio-lingual and disto-lingual surfaces of all teeth present, excluding third molars. Data were analysed cross-sectionally at each examination.Results: Over the period of the study, the prevalence of LCAL greater than or equal to1 and 2 mm ranged from 95-100%, whereas LCAL greater than or equal to3 mm ranged from 40-47%. The extent of LCAL greater than or equal to1 mm ranged from 76-86%. However, the extent of LCAL greater than or equal to2 mm was dramatically lower (10.5-12.7%), and LCAL greater than or equal to3 mm was uncommon (0.5-0.9%). Examining the number of subjects according to the number of sites affected above a threshold, showed that a small number of subjects have a large number of sites above threshold. Using Pearson's rank correlation coefficient a significant correlation (p

Previous studies have shown that the use of index teeth may underestimate the prevalence of chronic periodontitis in adults. However, there is little information on the effect of using index teeth to estimate the prevalence of early periodontitis in younger adults and the effect this may have on planning treatment needs and health care resources. The aim of this study was to compare full mouth examination with partial examination using index teeth in a group of young British males.One hundred subjects aged between 16 and 20 years (mean 17 years) on entry to the study were examined at baseline, 12 months later, and 30 months later. Lifetime cumulative attachment loss (LCAL)or =1 mm was measured on the mesio-buccal, disto-buccal, mesio-lingual, and disto-lingual surfaces of all teeth, excluding third molars. All data were entered into a database. The indices used to express LCAL were prevalence, defined as the percentage of subjects with LCALor =1 mm, 2 mm, or 3 mm, and extent, defined as the percentage of sites with LCALor =1 mm, 2 mm, or 3 mm. Two sets of index teeth were chosen to compare with full mouth recordings, Ramfjord index teeth and the Periodontal Index for Treatment (PIT) teeth.The prevalence of LCALor =1 mm was similar (approaching 100%) for the full mouth and both partial mouth recordings. However, as LCAL increased from a minimum of 1 to 3 mm, partial mouth recording resulted in an underestimation of the prevalence of disease. LCALor =2 mm was underestimated by up to 22% and LCALor =3 mm by up to 36%. The extent of LCAL was less affected by partial mouth recording, in that the percentage of sites with no sign of early attachment loss was underestimated by up to 11%. However, the percentage of sites with LCALor =1 mm and 2 mm were overestimated by 11% and, 7% respectively.These data indicate that the use of index teeth in epidemiological studies which include young adults may result in an underestimation of the prevalence of early periodontitis and an overestimation of the extent.

Background, aims: Conventional treatment of chronic periodontitis involves mechanical debridement of periodontal pockets. Recently, subgingival antimicrobials have been used adjunctively following such debridement. This 2-centre study compared the clinical effects of subgingival scaling (SRP) with SRP plus subgingival application of 25% metronidazole gel, Elyzol(R) (SRP+gel), in patients with chronic adult periodontitis.Method: Voluntary informed written consent was obtained from 45 subjects at the Eastman (mean age 46, range 34-63) and 43 subjects at RAF Halton (mean age 47, range 34-71) who participated in this blind, randomised split-mouth design study. All had at least 2 sites in each quadrant with probing pocket depth (PPD) greater than or equal to5 mm. PPD bleeding on probing (BOP), and clinical probing attachment levels (CAL) measured using a stent, were recorded at baseline and at 1, 3, 6 and 9 months posttherapy. After subgingival scaling of all quadrants, 2 quadrants were randomly selected to be treated with metronidazole gel.Results: A paired t-test on baseline values showed no bias between groups. Both treatments effectively reduced the signs of periodontitis. At each follow-up visit, reduction in PPD, CAL and BOP after the combined treatment was greater than for SRP alone. Paired t-tests showed that the improvement in the SRP+gel group was statistically significantly better (p 2 mm were greater for SRP+gel than for SRP alone.Conclusions: At the end of the study, the mean reductions for PPD were 1.0 mm (SRP) compared to 1.5 mm (SRP+gel), and for CAL they were 0.4 mm (SRP) compared to 0.8 mm (SRP+gel), with mean difference for CAL between treatments of 0.4+/-0.6 mm (95% confidence intervals of 0.3-0.6 mm). The combination therapy of SRP+gel was superior to the conventional treatment of SRP alone, and these differences were maintained for 9 months.

The present clinical trial was designed to evaluate the effects of a bioactive glass, Perioglas, in the treatment of periodontal intrabony defects. 20 patients, 23-55 years of age (44 sites), with intrabony defects completed the 1-year study. Teeth with furcation involvement were excluded. After completion of initial therapy, defects were randomly assigned to either a test or control procedure. Following flap reflection, root planing and removal of chronic inflammatory tissue in both groups, the test defects were restored with the bioactive glass particulate material. Mucoperiosteal flaps were replaced, sutured and a periodontal dressing was used. All the patients received postoperative antibiotics and analgesics and were seen at 1 week for suture removal. Follow-up was then carried out weekly and at 3 months, 6 months, 9 months and 1 year post-surgery. Plaque score, bleeding score, probing pocket depth (PPD), probing attachment level (PAL) and gingival recession were recorded at baseline, 3 months and 1 year. Standardised radiographs for computer-assisted densitometric image analysis (CADIA) were taken at baseline, immediately post-operatively and at 1 year. The CADIA data showed a significant increase (F-ratio: 15.67, p < 0.001) in radiographic density and volume between the defects treated with the Perioglas when compared to those treated with surgical debridement only. PPD and PAL showed significant improvements in both experimental and control sites, with a greater trend to improvement in the experimental sites. It was concluded that this bioactive glass is effective as an adjunct to conventional surgery in the treatment of intrabony defects.

Large amounts of biological samples are usually required to measure multiple components by the enzyme-linked immunosorbant assay. However, the amounts of many tissue extracts and fluids, including gingival crevicular fluid (GCF), are generally extremely small. The aim of this study was, therefore, to develop and validate a novel multiplex bead assay (MBA) to simultaneously measure a profile of healing-related mediators in the GCF of treated periodontal wounds. An MBA was developed and validated by assessment of assay selectivity, recovery, precision and sensitivity, using eight recombinant human growth mediators as assay standards. GCF samples were collected on paper strips from healing wound (test) and healthy unaffected (control) sites of 15 patients with periodontitis, seven days post-periodontal surgery. Each GCF sample was eluted and the levels of the mediators measured using the MBA and antibody pairs specific for angiopoietin-1, vascular endothelial growth-factor, bone morphogenetic protein-2, osteoprotegerin, tissue inhibitor of metalloprotease-1 (TIMP-1), basic fibroblast growth-factor, keratinocyte growth-factor, and platelet derived growth-factor. Less than 1.8% of cross-reactivity was observed between antibodies and the eight different analytes, for which the recovery was more than 85%. Mean intra- and inter-assay precision were within the acceptance criteria of 20% and 25%, respectively. Detection of all mediators was highly sensitive (or=70 ng/L) except for TIMP-1 (215 ng/L). Angiogenic factors were the most highly secreted in the GCF seven days post-surgery. This new MBA can simultaneously measure small amounts of eight different growth mediators in the GCF of healing periodontal wounds. It might also be a valuable tool for evaluating the components of wound fluids as a prognostic indicator of the success of therapeutic intervention.

Background and Objective: Family history is a primary diagnostic criterion for current classification of aggressive periodontitis (AgP). However, results of previous studies have shed controversy over the degree of familiarity of AgP and its possible inheritance mechanisms. The aims of this study were to estimate the percentage of affected relatives of AgP individuals, to analyse the disease phenotypes in relatives and to explore the distributions of genetic polymorphisms of interleukin-6 (IL-6) in AgP patients and in diseased and healthy relatives. Material and Methods: Patients with AgP were clinically examined and asked to provide relatives for examination. First-degree relatives were clinically and radiographically diagnosed. Blood samples were collected, DNA was extracted and analysis of single nucleotide polymorphisms of IL-6 (at positions −174, −1363 and −1480) by polymerase chain reaction was performed in patients and relatives. Results: Fifty-five AgP patients provided relatives for examination. A total of 100 first-degree relatives were assessed and 10 of them (10%) were found to have AgP. All relatives diagnosed with AgP had the same disease as the corresponding proband (localized AgP/localized AgP or generalized AgP/generalized AgP). The same IL-6 genotypes (−174 GG, −1480 CC) previously associated with AgP showed a tendency for association with AgP in relatives. Conclusion: This pilot study confirmed a relatively high risk for relatives of AgP patients to have AgP (10%). Genetic polymorphisms in the IL-6 gene may have an impact in aetiopathogenesis. This study provides a sample size calculation for a novel study design using healthy relatives as control subjects.

We recently reported an association between interleukin-6 (IL6) polymorphisms (SNPs) and haplotypes and aggressive periodontitis (AgP). The aim of this study was to investigate this association in a larger cohort of subjects, affected by either aggressive or chronic periodontitis. Five IL6 SNPs were analyzed in 765 subjects (167 generalized aggressive periodontitis, 57 localized aggressive, 310 chronic periodontitis and 231 periodontally healthy). Among Caucasians (n = 454) there were moderate associations for -1363T allele (p=0.011) and for -174GG and -1363GG genotypes with diagnosis of periodontitis (respectively, p = 0.044, OR = 1.6, 95% CI = 1.0-2.4, and p = 0.017, OR = 1.8. 95% CI = 1.1-2.8, adjusted for age, gender and smoking). Haplotypes containing the -174G>C, -1363G>T and -1480C>G polymorphisms were associated with diagnosis of periodontitis (p = 0.02). Subgroup analysis by disease phenotype showed associations for the localized AgP (LAgP) group and -1480C>G and -6106A>T SNPs (p=0.007 and 0.010, respectively). Among Caucasians the genotypes IL6 -1480 CC and -6106 TT increased the adjusted OR for LAgP (OR = 3.09 and 2.27, respectively). This study supports the hypothesis that 116 polymorphisms and haplotypes are moderately associated with periodontitis, possibly acting through influencing tissue levels of IL6. This association is stronger for LAgP than for other periodontal disease phenotypes. (C) 2008 Elsevier Ltd. All rights reserved.

Background: Interleukin-6 (IL-6) polymorphisms have been shown to affect IL-6 promoter activity. This study investigated the possible role of IL-6 genetic polymorphisms and haplotypes in the predisposition to aggressive periodontitis (AgP). Material and Methods: A case-control association study on 224 AgP patients and 231 healthy controls was performed in order to detect differences in genotype distributions of five single nucleotide polymorphisms (SNPs) located in the promoter region of the IL-6 gene. Results: The IL-6 - 1363 polymorphism was associated with a diagnosis of AgP in subjects of all ethnicities (p = 0.006, adjusted logistic regression). The - 1480 SNP was associated with LAgP in subjects of all ethnicities (p = 0.003). The - 1480 and - 6106 polymorphisms were associated with Localized AgP in Caucasians (n = 24) (p = 0.007 and 0.010, respectively). Haplotypes determined by the - 1363 and - 1480 polymorphisms were also associated with LAgP (p = 0.001) in Caucasians. Conclusions: This study supports the hypothesis of a link between IL-6 genetic factors and AgP and highlights the importance of two IL-6 polymorphisms (- 1363 and - 1480) in modulating disease phenotype and susceptibility.

Background and Aim: A cluster of metabolic factors defines a syndrome that predisposes to diabetes and cardiovascular disease. Chronic infections such as periodontitis might alter these individual metabolic factors and the systemic inflammatory burden. The aim of this study was to investigate the association between severe periodontitis and increase in inflammatory and metabolic risk factors for cardiovascular disease.Materials and Methods: We examined 302 patients with severe periodontitis and 183 healthy controls, and we collected a blood sample from each subject in order to investigate differences in inflammatory (leukocyte numbers and differential counts) and metabolic markers (lipids and glucose).Results: After correcting for differences in age, gender, smoking and ethnicity, periodontitis subjects exhibited a low-grade systemic inflammation (increased white cell counts, 1.10 +/- 1.02 x 10(9)/l, 95%CI 1.05-1.15, p=0.0001), dyslipidemia [lower high-density lipoprotein cholesterol, 1.14 +/- 1.03 mmol/l, 95%CI 1.08-1.20, p < 0.0001 and higher low-density lipoprotein cholesterol, 1.12 +/- 1.03, 95%CI 1.05-1.19, p < 0.0001) and increased non-fasting serum glucose levels (1.04 +/- 1.01 mmol/l, 95%CI 1.02-1.06, p=0.01) when compared with controls. The associations were confirmed in a subpopulation of Caucasian non-smokers. A trend for a dose dependent effect of the number of periodontal pockets on the tested inflammatory and metabolic markers was observed.Conclusions: These data suggest a possible link between severe generalized periodontitis, systemic inflammation and a dysmetabolic state in otherwise healthy individuals.

Growing evidence suggests that individual genetic susceptibility may influence the host’s response to infections. The aim of this project was to study whether gene polymorphisms of inflammatory markers are associated with the presence of viable periodontopathogenic bacteria. We extracted genomic DNA from 45 young adults diagnosed with generalized aggressive periodontitis to study Fc receptors, formyl peptide receptor, Interleukin-6, tumor necrosis factor-α, and vitamin D receptor polymorphisms. The presence and viable numbers of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis were determined by culture, and their identities confirmed by PCR. Multiple logistic regressions revealed that both Fcγ receptor and IL-6 -174 polymorphisms were associated with increased odds of detecting A. actinomycetemcomitans, P. gingivalis, and T. forsythensis after adjustment for age, ethnicity, smoking, and periodontitis extent. These findings support the hypothesis that complex interactions between the microbiota and host genome may be at the basis of susceptibility to aggressive periodontitis.

INTRODUCTION: Neutrophils (PMN) in aggressive periodontitis (AgP) patients have been reported to be hyperactive especially with regards to superoxide production. Polymorphisms in genes influencing PMN function have been proposed as candidate risk factors for AgP. The aim of this study was to test the association of specific gene polymorphisms affecting PMN functions with AgP. MATERIALS AND METHODS: Two hundred and twenty-four patients with confirmed diagnosis of AgP and 231 subjects with healthy periodontium took part in the study. A blood sample was collected from subjects and genotypes for p22phox (CYBA) NADPH oxidase, FP, Fcalpha and Fcgamma receptors were analysed in a blind fashion. RESULTS: The C242T p22phox NADPH oxidase T allele was significantly associated with AgP in a multiple logistic regression model adjusting for confounders, and this was observed for all subjects [p = 0.002, odds ratio (OR) = 1.87, 95% confidence interval (CI) = 1.27-2.83] and Caucasians (p = 0.009, OR=2.07, 95% CI = 1.20-3.59). Concomitant presence of C242T p22phox NADPH oxidase T allele and FcgammaRIIIb NA1 homozygosity was associated with the generalized AgP phenotype in Caucasians (p = 0.001, OR = 30.35, 95% CI = 3.81-241.97). CONCLUSIONS: C242T p22phox NADPH oxidase and FcgammaR polymorphisms may predispose to AgP through a modulation of neutrophil superoxide production.

There is strong evidence that genetic as well as environmental factors affect the development of periodontitis, and some suggestion that aggressive and chronic forms of the disease share the same genetic predisposition. This study addresses the hypothesis that there are both shared and unique genetic associations in these forms of periodontitis. A sample of 51 patients with aggressive disease, 57 patients with chronic disease, and 100 healthy controls was recruited for this study. Ten functional polymorphisms in 7 candidate genes were genotyped. The results show statistically significant (p ≤ 0.05) differences between genotype frequencies in aggressive and controls (IL-1B +3954 & IL-6 −174); chronic and controls (IL-6 −174 & VDR −1056); chronic and aggressive periodontitis (IL-1A −889); and periodontitis as a whole and controls (VDR −1056, TLR-4 399 & IL-6 −174). These results suggest that there are in fact both shared and unique genetic associations in aggressive and chronic periodontitis.

Background: Inflammation of periodontal tissues during postoperative wound healing is mediated by cell surface adhesion molecules. Soluble forms of these antigens have also been identified and shown to be important in immunoregulatory processes, but have previously not been investigated during periodontal repair and regeneration. The present study has examined the presence and possible changes in soluble intercellular adhesion molecule-1 (sICAM-1; CD54) and lymphocyte function-associated antigen-3 (sLFA-3; CD58) in gingival crevical fluid (GCF) following periodontal surgery. Methods: GCF samples were collected from four groups: 1) a guided tissue regeneration (GTR) test; 2) a GTR control, at least one complete tooth unit away from the periodontal defect; 3) a conventional flap (CF) surgery; and 4) a crown lengthening (CL). Sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of sICAM-1 and sLFA-3 in the GCF samples. Results: A marked increase in GCF volumes was found in all sites after surgery, although a persistent increase was associated only with the period of membrane retention at the GTR test sites. In addition, sICAM-1 and sLFA-3 were found in the GCF of healthy as well as diseased sites prior to treatment and the total amounts of both increased transiently following surgical intervention, especially sLFA-3. However, the concentrations of these GCF components, particularly sICAM-1, tended to decrease. Conclusions: The temporal decrease in the concentration of sICAM-1 and sLFA-3 in GCF may serve to enhance inflammatory reactions at surgically-treated periodontal sites, thereby limiting repair and regeneration in the periodontium. These soluble adhesion molecules may thereby be of potential therapeutic value and might also be useful markers for monitoring periodontal wound healing. J Periodontol 2005;76:526-533.

Objectives: Growth factors play a major part in wound healing, including in the periodontium. However, the presence of growth factors in gingival crevicular fluid (GCF) in humans during periodontal wound healing has not yet been determined. Our hypothesis is that such factors are present in GCF and that changes in their levels might be of value as a prognostic marker of wound-healing activity and therapeutic progress following periodontal surgery. The aim of this study was therefore to measure transforming growth factor-beta1 (TGF-beta1) in GCF collected from sites that have undergone guided tissue regeneration (GTR) and conventional flap (CF) surgery and to compare these with GCF collected from unaffected healthy sites.Materials and Methods: GCF samples were collected, using filter paper strips, at baseline (pre-surgical) and then at intervals up to 26 weeks from 16 patients undergoing GTR and from 11 patients undergoing CF surgery. After elution and acid treatment, TGF-beta1 levels were measured by ELISA.Results: Treatment of periodontal defect sites significantly reduced the mean probing pocket depth (PPD) and improved the mean lifetime cumulative attachment loss (LCAL). Average GCF volumes also significantly increased at all sites at 2 weeks post-surgery and thereafter declined to baseline levels, except at the GTR test sites that were still elevated at 7 weeks. TGF-beta1 could be detected in almost all GCF samples, and 2 weeks after surgery, the average levels increased two-fold at the surgically treated but not at the control sites, which remained unchanged.Conclusion: TGF-beta1 is readily detectable in GCF and increases transiently following periodontal surgery. This suggests that changes in the levels of this growth factor in GCF might be useful for monitoring the progress of periodontal repair and regeneration.

Background: The aim of this study was to investigate the longitudinal relationships between the outcome measurements of changes in lifetime cumulative attachment loss (cLCAL) and changes in probing depth (cPD) in relation to potential risk factors or other risk markers for periodontal disease progression from a cohort of 100 young males. In order to account for the hierarchical data structure, and to explore explicitly the site, tooth, and subject levels simultaneously, multilevel modeling was undertaken. Methods: The analyses were undertaken in two parts. Within a previous article, the absolute levels of disease were analyzed in relation to potential risk factors; within this article, changes in disease are analyzed in relation to these factors. Each analytical approach yielded substantively different insights. Results: Subject-level risk factors had limited predictive value for cLCAL/cPD throughout the 30-month observation period. Tooth position demonstrated a near linear relationship for both outcomes, with disease increasing from anterior to posterior teeth. Supragingival plaque had no significant effect on cLCAL/ cPD, while subgingival calculus and bleeding on probing were negatively associated with cLCAL/cPD. In contrast to the outcomes LCAL/PD, supragingival calculus had no significant protective effect on cLCAL/cPD. There was no significant influence of smoking in this cohort. Conclusions: This study provides, for a relatively young cohort, considerable insights into the factors associated with longitudinal patterns of early-life periodontal disease at all levels of the natural hierarchy of sites within teeth within subjects. Furthermore, it is demonstrated how multilevel modeling can provide considerable insight into some of the inconsistencies and controversies found in the previous periodontal literature. J Periodontol 2004;75:137-145.

Statistical analyses of periodontal data that average site measurements to subject mean values are unable to explore the site-specific nature of periodontal diseases. Multilevel modeling (MLM) overcomes this, taking hierarchical structure into account. MLM was used to investigate longitudinal relationships between the outcomes of lifetime cumulative attachment loss (LCAL) and probing depth (PD) in relation to potential risk factors for periodontal disease progression.One hundred males (mean age 17 years) received a comprehensive periodontal examination at baseline and at 12 and 30 months. The resulting data were analyzed in two stages. In stage one (reported here), the absolute levels of disease were analyzed in relation to potential risk factors; in stage two (reported in a second paper), changes in disease patterns over time were analyzed in relation to the same risk factors. Each approach yielded substantially different insights.For absolute levels of disease, subject-level risk factors (covariates) had limited prediction for LCAL/PD throughout the 30-month observation period. Tooth position demonstrated a near linear relationship for both outcomes, with disease increasing from anterior to posterior teeth. Sites with subgingival calculus and bleeding on probing demonstrated more LCAL and PD, and supragingival calculus had an apparently protective effect. Covariates had more "explanatory power" for the variation in PD than for the variation in LCAL, suggesting that LCAL and PD might be generally associated with a different profile of covariates.This study provides, for a relatively young cohort, considerable insights into the factors associated with early-life periodontal disease and its progression at all levels of the natural hierarchy of sites within teeth within subjects.

Previously, burst and linear theories for periodontal disease progression were proposed based on different but limited statistical methods of analysis. Multilevel modeling provides a new approach, yielding a more comprehensive model. Random coefficient models were used to analyze longitudinal periodontal data consisting of repeated measures (level 1), sites (level 2), teeth (level 3), and subjects (level 4). Large negative and highly significant correlations between random linear and quadratic time coefficients indicated that subjects and teeth with greater-than-average linear change experienced decelerated variation. Conversely, subjects and teeth with less-than-average linear change experienced accelerated variation. Change therefore exhibited a dynamic regression to the mean at the tooth and subject levels. Since no equilibrium was attained throughout the study, changes were cyclical. When considered as a multilevel system, the "linear" and "burst" theories of periodontal disease progression are a manifestation of the same phenomenon: Some sites improve while others progress, in a cyclical manner.

Expanded polytetrafluoroethylene (ePTFE) barrier membranes have been widely used for guided tissue regeneration (GTR) of the human periodontal ligament (PL). However, the precise cellular and molecular events involved in the re-growth of the new tissue are still unclear.Retrieved membranes and the newly-regenerated soft tissue (RT) underlying the membranes were used to examine the cells associated with GTR compared with normal human PL and gingival cells. Flow cytometry (FCM) was used, for the first time, to analyze the spindle-shaped fibroblast-like cells which were adherent to these membranes and the cells which grew out of the RT.The results showed that the membrane-associated (M) cells had the lowest rate of proliferation and appeared to be larger and more granular than the other types of cell. Moreover, both the M- and RT-derived cells were found to express higher levels of the extracellular matrix (ECM) proteins collagen type 1, fibronectin, tenascin, and decorin. In addition, evidence based on FCM profiles identified distinct sub-populations of GTR cells in which fibronectin expression was markedly up-regulated compared with normal PL cells and which also differed in size and granularity.The results of this study show that cells associated with GTR barrier membranes and with the underlying tissue appear to have distinct phenotypic and functional activities consistent with the production of new periodontal connective tissue and periodontal regeneration.

To introduce the concepts of random coefficient multilevel models through an application to periodontal research data.Multilevel models with random coefficients are illustrated using periodontal data that comprise four levels: repeated measurements at level-1, sites at level-2, teeth at level-3, and subjects at level-4. The study explores random coefficient models--where random variation occurs about explanatory variable coefficients. Outcomes considered are lifetime cumulative attachment loss and pocket probing depth.The study data were taken from a survey of periodontal disease involving 100 white male trainee engineers aged between 16 and 20 entering the apprentice training school at the Royal Air Force-Halton, UK.The application of multilevel modelling to longitudinal data provides a new way of exploring old problems. The multilevel random coefficient models provide an opportunity to examine the 'linear' and 'burst' theories of periodontal disease progression, leading to the postulation that both can be unified within the multilevel framework.The multilevel methodology illustrates how advances in the understanding of oral health can be achieved with the advent of new statistical methods

To explain the theory of multilevel modelling and demonstrate its application in the analysis of dental research data.Multilevel modelling was introduced using dental data comprising four levels: repeated measurements at level-1, sites at level-2, teeth at level-3, and subjects at level-4. Variance components models (which have no explanatory variables) were evaluated for all outcome measures. Explanatory variables were added to the models with outcomes for both lifetime cumulative attachment loss and pocket probing depth. Salient features of the multilevel models were discussed.Research data were obtained from a longitudinal survey of periodontal disease conducted on 100 white male trainee engineers aged between 16 and 20 years entering the apprentice training school at Royal Air Force Halton, England.The statistical methods revealed that periodontal measures demonstrate considerable variation at all levels of the multilevel structure. Models for lifetime cumulative attachment loss and pocket probing depth illustrated that risk factors operated at more than one level. Supragingival calculus was a risk factor at the subject-level (subjects experiencing more sites with the condition had greater attachment loss and greater pocketing) whilst there was apparently a protective effect occurring at the site (sites with the condition had less attachment loss and less pocketing).This study demonstrates that multilevel modelling is a more powerful research tool than single-level techniques for the analysis of hierarchical dental data. Researchers using these techniques are well equipped to analyse complex hierarchical data structures, such as those often found within dentistry.

Osteocalcin (Oc) and the collagen cross-links pyridinoline (Pyr) and deoxypyridinoline (dPyr) are used as markers of bone turnover in metabolic bone diseases. The aims of this study were: 1) to establish if Oc, Pyr and dPyr can be detected in GCF and 2) using the orthodontic tooth movement model of alveolar bone resorption to evaluate GCF levels of osteocalcin and these collagen cross-links as markers of bone breakdown. Plaque, colour and bleeding indices, probing measurements and GCF samples were collected at two sites in each of 20 adolescents, during 4 stages of fixed appliance therapy: (1) prior to appliance fit, (2) post appliance fit, (3) during active retraction of the maxillary canines, (4) during retention. GCF was collected onto filter paper strips and the volume determined by weighing. An ELISA kit was used for the detection of osteocalcin, whereas Pyr and dPyr were assayed using high performance liquid chromotography (HPLC). Wilcoxon signed ranks test and Bonferroni correction revealed statistically significant increases in plaque (p= 0.012), GCF volume (p=0.024) and osteocalcin concentration (p=0.012), between stages 1 and 2. There were no statistically significant differences between the other variables at this stage or between any of the variables at stages 2 and 3, or between stages 3 and 4. All but 3 of the GCF samples yielded detectable osteocalcin, with large site and subject variation. The median values of osteocalcin and osteocalcin concentration of all the samples were 87.5 pg and 66 pg/microl, with a range of 0-1,248 pg, 0-1,572 pg/microl. The detection of osteocalcin in GCF during every stage, the wide variation between subjects, and the lack of a consistent pattern related to stages of orthodontic treatment, suggests that osteocalcin may merely be a constituent of GCF associated with the developing dentition, which would reduce its potential as a marker of bone turnover in this group. None of the 16 GCF samples analysed for Pyr and dPyr gave a positive result. This study confirms that fitting an orthodontic appliance results in plaque accumulation and increased gingival inflammation, and that GCF volume is the most sensitive indicator of that inflammation. Osteocalcin was detected in GCF collected from adolescents, whereas Pyr and dPyr could not be detected. Further work is required to establish whether GCF osteocalcin levels can be used as a marker of bone turnover, and whether improvements in the sensitivity of detecting Pyr & dPyr make further study of these promising bone markers worthwhile.

Gingival crevicular fluid (GCF) sampling was performed on 2 occasions separated by 1 year, at 2 sites in the mouths of 102 male adolescents, mean age 17.85 years. Samples were collected onto 5 filter paper strips which were sequentially applied to the mouth of the crevice over a 9-min collection period. Volume and flow rates of GCF were determined for each site and were compared with clinical measurements of plaque, gingival colour, bleeding, gingival index (GI) and pocket depth, using a general linear models (GLM) procedure. While the initial volume of GCF showed no association with any clinical measurement, there was an association between flow rate of GCF and gingival colour. The volume of GCF collected in the final, 5th sample was associated with the GI. The sample site strongly influenced all measures of GCF volume. It is proposed that the flow rate of GCF may be a better indicator of gingival inflammation than the more imprecise clinical assessments of inflammation, since GCF flow rates more precisely reflect changes in tissue permeability. The association between the final sample, collected after 9 min, and clinical measurements, was probably a reflection of the association between clinically-detectable inflammation and the susceptibility of the site to mild irritation.