Anti-apolipoprotein A-1 autoantibodies as biomarker for atherosclerosis burden in patients with periodontitis.

BACKGROUND AND OBJECTIVE: Anti-apolipoprotein A-1 (anti-apoA-1) IgG is a potential marker of atherosclerotic plaque vulnerability and cardiovascular complications. In patients with periodontitis the presence of anti-apoA-1 IgGs in serum and their association with atherosclerosis is unknown. MATERIAL AND METHODS: One-hundred and thirty subjects with periodontal disease and 46 healthy subjects, matched for age and gender, participated in this study. Anti-apoA-1 IgG, high-sensitivity C-reactive protein (hsCRP) and matrix metalloproteinase (MMP) -2, -3, -8 and -9 were measured in serum samples. An ankle-brachial index (ABI) value below 1.11 served as a surrogate marker of atherosclerosis. Predictive accuracies of biomarkers for abnormal ABI were determined using receiver-operating characteristics curves and logistic regression analyses. RESULTS: Compared with healthy controls, periodontitis patients showed lower median ABI values (1.10 vs. 1.15; p < 0.0001), a higher prevalence of anti-apoA-1 IgG positivity (23.8% vs. 6.5%; p = 0.009) and higher concentrations of hsCRP (1.62 mg/L vs. 0.85 mg/L; p = 0.02) and MMP-9 (435 µg/mL vs. 283 µg/mL; p < 0.0001). In patients younger than 50 years of age (n = 66), anti-apoA-1 IgG was found to be the best predictor for an abnormal ABI (area under the curve = 0.63; p = 0.03). Anti-apoA-1 IgG positivity increased the risk of having an abnormal ABI (odds ratio = 4.20; p = 0.04), independently of diabetes, smoking and body mass index. CONCLUSIONS: Anti-apoA-1 IgG positivity and atherosclerosis, as reflected by abnormal ABI, were more prevalent in periodontitis patients than in age- and gender-matched controls. In younger periodontitis patients, anti-apoA-1 IgG was found to be the best predictor of atherosclerosis burden.

Anti-infective surgical therapy of peri-implantitis. A 12-month prospective clinical study.

AIM: The aim of this prospective cohort study was to evaluate an anti-infective surgical protocol for the treatment of peri-implantitis. MATERIALS AND METHODS: Thirty-six implants in 24 partially dentate patients with moderate to advanced peri-implantitis were treated using an anti-infective surgical protocol incorporating open flap debridement and implant surface decontamination, with adjunctive systemic amoxicillin and metronidazole. Treatment outcomes were assessed at 3, 6 and 12 months. Patient-based statistical analyses using multiple regression analyses were performed. RESULTS: There was 100% survival of treated implants at 12 months. At 3 months, there were statistically significant (P < 0.01) reductions in mean probing depths (PD), Bleeding on Probing (BoP) and suppuration. The greater the mean PD at baseline, the greater the PD reduction at 3 months. At 3 months, there was also a significant mean facial mucosal recession of 1 mm (P < 0.001). All these changes were maintained at 6 and 12 months. At 12 months, all treated implants had a mean PD < 5 mm, while 47% of the implants had complete resolution of inflammation (BoP negative). At 12 months, 92% of implants had stable crestal bone levels or bone gain. There were no significant effects of smoking on any of the treatment outcomes. CONCLUSIONS: For the treatment of peri-implantitis, an anti-infective protocol incorporating surgical access, implant surface decontamination and systemic antimicrobials followed by a strict postoperative protocol was effective at 3 months with the results maintained for up to 12 months after treatment.

Myeloid-related protein (MRP8/14) expression in gingival crevice fluid in periodontal health and disease and after treatment.

BACKGROUND AND OBJECTIVE: Myeloid-related protein (MRP8/14) and its subunits are biomarkers of inflammation. The present study evaluated whether gingival crevice fluid levels of these markers discriminate periodontitis from healthy sites in patients with chronic periodontitis or diseased from healthy subjects, and whether these biomarkers detect longitudinal changes after therapy. MATERIAL AND METHODS: Levels of MRP8/14, MRP14 and total protein were quantified in 19 periodontitis patients before non-surgical periodontal therapy, after 3 and 6 mo of treatment, and were measured once in 11 periodontally healthy subjects. In total, diseased subjects contributed 59 sites with probing depths >4 mm (PP) and 21 sites <4 mm (PH); healthy subjects contributed 91 sites (HH). RESULTS: Overall, in diseased subjects, MRP8/14, MRP14 and total protein were not significantly different between PP and PH sites. However, at baseline, MRP8/14 and total protein had significantly higher values at sites in periodontally diseased than in healthy subjects. Clinical improvement was associated with a significant decrease of MRP8/14 and MRP14 from baseline to month 6 in PP sites. Interestingly, a similar decrease was observed in PH sites for all three markers. At 6 mo, however, levels of MRP8/14 and protein in PP and PH sites of patients were still significantly higher than in healthy subjects. CONCLUSION: Gingival crevice fluid levels of MRP8/14 did not differentiate between clinically diseased and healthy sites in patients with chronic periodontitis. However, this marker was elevated in periodontally diseased compared with healthy subjects, and its values decreased following therapy. MRP8/14 may be used to monitor the response to treatment.

Cytokines in gingival crevicular fluid of adolescents and young adults.

BACKGROUND/AIM: The purpose of this study was to compare the levels of the cytokines interleukin-1beta (IL-1beta), IL-4, and IL-8 in the gingival crevicular fluid (GCF) of adolescents and young adults. METHODS: Twenty-five adolescents aged between 14 and 16 years (Group A) and 20 periodontally healthy young adults aged between 25 and 35 years (Group B) were selected from two private dental clinics limited to pedodontics and periodontics respectively in Piraeus Greece. All subjects were systemically healthy. Clinical examination included probing pocket depth (PPD), presence or absence of plaque, and bleeding on probing (BOP). GCF was collected from four sites per subject. IL-1beta, IL-4, and IL-8, measured as total amounts (pg/30 s), were evaluated in 180 samples using a commercially available sandwich enzyme-linked immunosorbent assay. RESULTS: IL-1beta mean levels of Groups A and B were adjusted for BOP and PPD. Differences of IL-1beta mean levels between the two age groups were statistically significant (F = 50.245, P < 0.001) in favour of Group A. Adolescents showed statistically significantly lower mean levels of IL-4 than young adults in the presence of BOP (F = 10.690, P = 0.001). There was no statistically significant difference between adolescents and adults for the means of IL-8 adjusted for BOP and plaque presence (F = 2.032, P = 0.161). CONCLUSIONS: Within the limits of this study the differences reported in mean levels of IL-1beta and IL-4 may be attributed to the different age status.

Effect of initial periodontal therapy on gingival crevicular fluid cytokine profile in subjects with chronic periodontitis.

Cytokines are thought to play an important role in the pathogenesis of periodontal disease. Because periodontal disease is known for its inhomogeneous distribution within the dentition, it is unclear to what extent the detection of various cytokines at different sites correlates with presence of disease. We evaluated whether levels of 12 cytokines in gingival crevicular fluid (GCF) discriminated periodontally diseased sites from healthy ones, or periodontally diseased persons from healthy ones, and assessed the impact of nonsurgical periodontal therapy on these readings. This study included 20 periodontally healthy persons (H) and 24 patients with chronic periodontitis (P). In every participant, we measured the plaque index, gingival index, probing pocket depth (PD), bleeding on probing, and recession at six sites of every tooth. GCF was collected with Durapore® filter strips from two healthy sites (PD<4 mm; HH) in group H, and from two periodontally diseased sites (PD≥5 mm; PP) and two periodontally healthy sites (PD≤3 mm; PH) in group P. The periodontally diseased participants underwent comprehensive nonsurgical periodontal therapy including deep scaling and root planing under local anesthesia. In these participants, GCF samples were again collected at the same sites 1 and 3 months after therapy. Twelve cytokines (il-1ß, il-1ra, il-6, il-8, il-17, b-fgf, g-csf, gm-csf, ifn-γ, mip-1ß, vegf, and tnf-α) were assessed using the Bio-Plex suspension array system. Mean plaque index, gingival index, bleeding on probing, PD, and recession were significantly different between groups H and P. Differences between PP and PH sites were not significant for any of the cytokines. Il-1ra, il-6, il-17, b-fgf, gm-csf, mip-1ß, and tnf-α differed significantly between HH sites and both PH and PP sites, whereas il-8 was significantly higher only at PP sites. Periodontal treatment increased gm-csf and decreased il-1ra levels in PP sites. Il-1ra, il-6, il-8, il-17, b-fgf, gm-csf, mip-1ß, and tnf-α identified patients with chronic periodontitis, rather than diseased sites, suggesting a generalized inflammatory state that is not limited to clinically diseased sites only.

Enamel matrix derivative and systemic antibiotics as adjuncts to non-surgical periodontal treatment: biologic response.

BACKGROUND: Short-term clinical observations suggest an anti-inflammatory effect of enamel matrix derivative (EMD). The purpose of this study was to evaluate the anti-inflammatory capacity of EMD, used as an adjunct to non-surgical periodontal treatment of deep lesions in chronic periodontitis patients, by monitoring inflammatory markers in gingival crevicular fluid (GCF). METHODS: Sixteen subjects were randomly assigned to treatment with EMD or placebo in contralateral dentition areas. Half of the subjects received 250 mg metronidazole and 375 mg amoxicillin three times a day for 7 days; the other half received a placebo. GCF samples were collected from one interproximal lesion in each of the contralateral quadrants before treatment and after 10 days and 2, 6, and 12 months. Total protein content was determined according to the Bradford method. Myeloid-related protein (MRP) 8/14 and interleukin (IL)-1beta were analyzed quantitatively by enzyme-linked immunosorbent assay (ELISA), and elastase activity was determined using a low molecular weight fluorogenic substrate. RESULTS: No significant differences were observed between sites treated with or without EMD for any biochemical parameter. Two months after treatment, subjects treated with antibiotics exhibited less clinical signs of inflammation. Furthermore, these subjects had lower MRP 8/14 levels only at day 10 compared to those receiving the placebo. For total protein, IL-1beta, and elastase, no statistically significant differences were noted for subjects with or without antibiotic therapy at any time point. CONCLUSIONS: Improved healing of the soft tissues has been noted clinically in non-surgically treated sites in subjects treated with antibiotics. The expression of inflammatory mediators in GCF corroborated this finding only in part. EMD did not seem to further affect the expression of inflammatory mediators.

Systemic Biomarkers in 2-Phase Antibiotic Periodontal Treatment: A Randomized Clinical Trial.

Accumulating evidence suggests that periodontal infections may have an impact on systemic health. In patients with untreated periodontitis, very high values for several inflammatory markers in serum are expressed simultaneously. We investigated to what extent these peak values change after nonsurgical and surgical periodontal treatment, with adjunctive antibiotics administered during the first or the second treatment phase. In a single-center, randomized, placebo-controlled, and double-masked clinical trial, 80 patients with chronic or aggressive periodontitis were randomized into 2 treatment groups: group A, receiving systemic amoxicillin and metronidazole during the first, nonsurgical phase of periodontal therapy (phase 1), and group B, receiving the antibiotics during the second, surgical phase (phase 2). Serum samples were obtained at baseline (BL), 3 mo after phase 1 (M3), and 6 and 12 mo after phase 2 (M6, M12). Samples were evaluated for 15 cytokines and 9 acute-phase proteins using the Bio-Plex bead array multianalyte detection system. For each analyte, peak values were defined as greater than mean +2 SD of measurements found in 40 periodontally healthy persons. Sixty-six patients showed a peak value of at least 1 analyte at BL. At M12, the number of these patients was only 36 (P = 0.0002). This decrease was stronger in group A (BL: 35, M12: 19, P = 0.0009) than in group B (BL: 31, M12: 17, P = 0.14). Twenty patients displayed peak values of at least 4 biomarkers at BL. The nonsurgical therapy delivered in the first phase reduced most of these peaks (group A, BL: 9, M3: 4, P = 0.17; group B, BL: 11, M3: 2, P = 0.01), irrespective of adjunctive antibiotics. The reductions obtained at M3 were maintained until M12 in both groups. Initial, nonsurgical periodontal therapy reduced the incidence of peak levels of inflammatory markers. Antibiotics and further surgical therapy did not enhance the effect (Clinicaltrials.gov NCT02197260).

Development and applications of injectable poly(ortho esters) for pain control and periodontal treatment.

Poly(ortho esters) with a low glass transition temperature are semi-solid materials so that therapeutic agents can be incorporated at room temperature, without the use of solvents, by a simple mixing procedure. When molecular weights are limited to < 5 kDa, such materials are directly injectable using a needle size no larger than 22 gauge. Somewhat hydrophilic polymers can be produced by using the diketene acetal 3,9-diethylidene-2,4,8,10-tetraoxaspiro[5.5]undecane and triethylene glycol (TEG), while hydrophobic materials can be produced by using the diketene acetal and 1,10-decanediol. Molecular weight can be reproducibly controlled by using an excess of the diol, or by use of an alcohol that acts as a chain-stopper. Erosion rates can be controlled by varying the amount of latent acid incorporated into the polymer backbone. Toxicology studies using the TEG polymer have been completed and have shown that the polymer is non-toxic. Toxicology studies using the decanediol polymer are underway. Development studies using the TEG polymer aimed at providing a sustained delivery of an analgesic agent to control post-surgical pain are under development and human clinical trials using the decanediol polymer for the treatment of periodontitis are also underway.

Isolation of periodontal species after guided tissue regeneration.

One advanced localized periodontal lesion in each of 10 patients was treated using the guided tissue regeneration procedure. Six weeks after placement of expanded polytetrafluoroethylene (ePTFE) membranes, microbial samples were taken from each treated site and the membranes were removed. Six weeks later the cases were re-evaluated. They had all healed successfully with varying amounts of gain of clinical attachment. Gram-negative, anaerobic rods were found in all samples and made up 31% of all organisms cultivated. In 1 patient, Porphyromonas gingivalis was found in a proportion of 17.5%. Six of the other 9 patients harbored Prevotella intermedia (mean proportion 21.3%) and 6 Prevotella melaninogenica (6.8%). Fusobacterium and Capnocytophaga were also frequently found. The results demonstrate that ePTFE membranes are frequently colonized by periodontal microorganisms. The importance of bacterial colonization on clinical success is presently not known. Further studies are needed to determine the effect of the presence or absence of putative pathogens during guided tissue regeneration.

Actinobacillus actinomycetemcomitans in adult periodontitis. I. Topographic distribution before and after treatment.

The aim of this investigation was to study the topographic distribution of Actinobacillus actinomycetemcomitans in patients with adult periodontitis before and after mechanical periodontal treatment (repeated oral hygiene instructions, systematic deep scaling, and root planing). In 10 A. actinomycetemcomitans-positive patients, subgingival microbial samples were obtained from the mesial and distal aspect of every tooth (38 to 56 sites per patient, 479 sites in total) before and one month after treatment. The samples were cultured on TSBV agar. A. actinomycetemcomitans was identified based on phenotypical and serological criteria. A. actinomycetemcomitans was present in 40% of the samples taken before and in 23% of the samples taken after treatment. Before treatment, the frequency of A. actinomycetemcomitans-positive samples per patient was wide spread and ranged from 7 to 90%. After treatment, two patterns of A. actinomycetemcomitans distribution could be recognized: the majority of the patients showed only a limited percentage of positive samples and yielded less than 10(5) A. actinomycetemcomitans. In three subjects, however, relatively high numbers of positive sites were still present, and many of these positive sites showed high A. actinomycetemcomitans counts. Logistic multiple regression showed the presence of A. actinomycetemcomitans before treatment depended strongly on the individual and was significantly associated with probing depth (P < 0.001) and bleeding upon sampling (P = 0.07). The highest chance of detecting A. actinomycetemcomitans existed in deep pockets which bled upon sampling. After treatment, there was a strong individual influence and an influence of probing depth (P < 0.001). The highest chance of detecting A. actinomycetemcomitans existed in residual pockets in the range of 5 mm.

Actinobacillus actinomycetemcomitans in adult periodontitis. II. Characterization of isolated strains and effect of mechanical periodontal treatment.

Ten systemically healthy subjects (ages 28 to 60 years) with untreated moderate to severe periodontal disease and evidence of presence of A. actinomycetemcomitans underwent standard mechanical periodontal treatment consisting of oral hygiene instruction and systematic deep scaling and root planing. Before, and 4 to 5 weeks after treatment, clinical measurements and separate subgingival microbiological samples were taken from the mesial and distal aspect of every tooth, with the exception of the third molars. A. actinomycetemcomitans could still be detected in all patients after treatment. In 9 of the 10 patients, all tested isolates from both examinations were of a single type. Two patients carried serotype a; 2 serotype b; 2 serotype c; and 1 serotype e. Two individuals showed only non-typeable isolates lacking serotype a, b, c, d, or e specific antigens. Another subject was colonized by serotype c and, in addition, yielded a non-typeable isolate. Persistence of A. actinomycetemcomitans after treatment was significantly correlated with the frequency of A. actinomycetemcomitans before treatment (P < 0.001) and the mean probing depth before treatment (P < 0.05). No serotype-specific patterns of treatment outcome could be recognized. The analysis of the site specific effect of treatment showed a significant relationship between post treatment levels of A. actinomycetemcomitans and both probing depth reduction as well as attachment gain. Individuals showing evidence of A. actinomycetemcomitans in a multitude of sites appeared to be more difficult to treat than patients with few positive sites only. Within such individuals, the deeper pockets showed the greater resistance to eradication of A. actinomycetemcomitans.

Persistence patterns of Porphyromonas gingivalis, Prevotella intermedia/nigrescens, and Actinobacillus actinomyetemcomitans after mechanical therapy of periodontal disease.

BACKGROUND: The aim of this study was to determine the distribution patterns of Porphyromonas gingivalis, Prevotella intermedia/nigrescens, and Actinobacillus actinomycetemcomitans in periodontitis patients after standard mechanical periodontal therapy, and to determine factors increasing the odds to detect these target organisms in treated sites. METHODS: Eight hundred fifty-two (852) separate subgingival microbial samples were taken from the mesial and distal aspects of every tooth in 17 patients. Target organisms were identified culturally. RESULTS: The 3 microorganisms showed different persistence patterns: P. gingivalis was detected in a high percentage of subjects (59%), but in a low proportion of sites (5.4%). P. intermedia/nigrescens was detected in all subjects except one, and in 40.6% of the tested sites. Only 5 subjects were A. actinomycetemcomitans positive, but 2 of them showed a very high number of positive sites (44% and 75%, respectively). A highly significant relationship was found between a subject's tendency to bleed upon sampling and the number of P. intermedia/nigrescens-positive sites. A significant portion of the variation in frequency of persisting P. gingivalis could be explained by the frequency of persisting pockets deeper than 4 mm. No similar relationship could be established between clinical parameters and A. actinomycetemcomitans. On a site level, the odds of detecting P. gingivalis increased by a factor of 2.47 (P= 0.0001) for every millimeter of residual probing depth; the odds of detecting P. intermedia/nigrescens increased by a factor of 1.84 (P= 0.0001). CONCLUSIONS: If, after standard mechanical periodontal therapy, a large number of sites continue to bleed, one may expect an increased number of sites positive for P. intermedia/ nigrescens. If many deep pockets persist, a greater number of P. gingivalis-positive sites can be expected.

Bioerodible injectable poly(ortho ester) for tetracycline controlled delivery to periodontal pockets: preliminary trial in humans.

The semisolid consistency of poly(ortho esters) (POEs) containing tetracycline free base allows direct injection in the periodontal pocket and shows sustained and almost constant in vitro release in phosphate buffer, pH 7.4 at 37 degrees C, for up to 14 days. Total polymer degradation concomitant with drug release was obtained. Formulations containing 10% or 20% (wt/wt) tetracycline were evaluated in a panel of 12 patients suffering from severe and recurrent periodontitis. In the first trial including 6 patients, single-rooted teeth and molar teeth with furcations were treated immediately after scaling and root planing. Patients tolerated both formulations well, experienced no pain during application, and showed no signs of irritation or discomfort during the observation period. However, retention of the formulation was minimal in this first study. An improved clinical protocol followed in the second study (stopping bleeding after scaling and root planning) prolonged the retention of the formulations in the inflamed periodontal pockets. For up to 11 days, tetracycline concentrations in the gingival crevicular fluid were higher than the minimum inhibitory concentration of tetracycline against most periodontal pathogens.

[Do new diagnostic tests improve the differential diagnosis and therapy of periodontitis? A review]. / Verbessern neue diagnostische Tests die Differentialdiagnose und Therapie der Parodontitis? Eine Ubersicht.

New diagnostic tools give access to information that was previously not available by classical clinical means. This includes methods for the determination of the composition of the microbial flora or for monitoring specific reactions of host tissues to the accumulation of plaque. DNA-probes, monoclonal antibodies and enzyme assays have already been advertised and are sold to dentists. The practical value of new tests depends upon the possibility to improve the efficacy of treatment based on previously inaccessible information. This paper reviews frequently raised clinical questions, the possible answers coming from new diagnostica, and the potential impact of this knowledge on periodontal therapy.

Local differences in the newly formed crevicular microbiota.

Subgingival plaque samples were taken from upper and lower right canines, premolars, and first molars (distal, mid-buccal and lingual) in 10 periodontally healthy individuals after four days without oral hygiene. The samples were examined in the darkfield microscope. Counts and proportions of morphotypes were related to the location of the sampled sites. The influence of site location on the distribution of morphotypes was studied by multiple regression analysis. Inter-individual differences were significant for the proportions of cocci, nonmotile rods and fusiform organisms (p less than 0.001), the influence of site location was not. Significant differences (p less than 0.05) were found between total bacterial counts of samples taken from different sites: Distal-buccal samples contained more bacteria than did mid-buccal samples, mid-buccal ones contained more than lingual samples; samples from more posterior teeth contained more bacteria. The experiment was repeated after 5 weeks. The intertest agreement (kappa) for the presence of the different morphotypes ranged between 0.13 and 0.30. Coefficients of variation of the proportions of morphotypes determined in repeated samples were between 1.35 and 3.13.

[Quality of water of dental units. A survey in the region of Berne, Switzerland]. / Wasserqualität in zahnärztlichen Behandlungseinheiten: Eine Standortbestimmung in der Region Bern.

The aim of this study was to assess the microbial contamination of the water from 175 dental units, which had not been in use for at least 12 hours, and to determine the usefulness of rinsing the water lines for 3 minutes. Only 10% of all units fulfilled all criteria for drinking water. Frequently the total bacterial counts markedly exceeded 100 CFU/ml. In all cases the rinsing decreased bacterial loads significantly. Bacterial contamination of water lines increased with the time the dental units remained unused. It is recommended to rinse dental unit water lines daily for at least 3 minutes before working on patients.

Wolinella recta and spirochetes in relation to alteration of probing depth in a population without regular dental care.

The purpose of this study was to determine the occurence of spirochetes and Wolinella sp. in untreated, nonselected subjects and to relate the presence of these organisms to changes in probing depth over a period of 180 days. Clinical parameters were recorded and subgingival microbial samples were taken mesiobuccally on the upper first molars of 120 subjects (mean age 37 +/- 14 years) with a history of no regular dental care. The number of motile rods and spirochetes were determined in the darkfield microscope. The samples were cultured anaerobically on a selective medium which favoured the growth of Wolinella sp. The grown microorganisms were submitted to a number of tests for the identification of W. recta. Of the 172 sites investigated longitudinally, 73 harboured motile rods, 45 harboured W. recta and 88 harboured spirochetes. 13 sites showed an increase in probing depth of greater than or equal to 2 mm. A multiple regresssion analysis revealed no statistically significant correlation between the numbers of W. recta at baseline and the likelihood of breakdown during the observation period. The number of spirochetes at baseline, however, was significantly related to the changes of probing depth (p greater than 0.001). It was concluded that spirochetes were more closely related to changes of probing depth in this population than the number of W. recta organisms present.