Influence of Anti-Infective Periodontal Therapy on Subgingival Microbiota Evaluated by Chair-Side Test Compared to qPCR-A Clinical Follow-Up Study.

A chair-side test (CST) for five periodontal pathogens (Aggregatibacter actinomycetemcomitans, A.a.; Porphyromonas gingivalis, P.g.; Prevotella intermedia, P.i.; Treponema denticola, T.d.; Tannerella forsythia, T.f.) was compared with qPCR in a previous clinical study on 100 periodontitis patients at first diagnosis (T0). Following non-surgical treatment alone (SRP) or in combination with systemic or local antibiotics, 74 patients (57.4 ± 13.5 years) were again tested at the same sites from 14 to 24 months after T0. Bacterial elimination (%; compared to T0) was determined for each single species and compared between both test systems. In all patients, all five pathogens could not be fully eliminated regardless of therapy or test method. Tested with CST, the mean elimination ranged from 90% for SRP + Amoxicillin/Metronidazole to 59.13% for SRP only. The corresponding qPCR values were 30% and 29.6%. Only A.a. was eradicated in 100% by SRP + Amoxicillin/Metronidazole tested by CST, and it was 80% when qPCR was the test method. CST agreed with qPCR in 98.7% in the detection of A.a., and 74.3%, 78.4%, 73.0%, and 48.7% for P.g., P.i., T.d., and T.f., respectively. Neither conventional treatment nor the additional use of antibiotics-even with the correct indication-could completely eradicate the tested pathogens or prevent pocket reinfection

Clinical evaluation of a newly developed chairside test to determine periodontal pathogens.

BACKGROUND The subgingival microbiota as well as determination of markers such as associated pathogens is still in the focus of dental research. The aim of this controlled clinical trial was to determine clinical applicability of a newly developed chairside bacterial test (CST) for the most relevant periodontal pathogens. METHODS Within 125 participants (100 with periodontitis, 25 healthy) two sulcus fluid samples each were collected and pooled for further analysis. Samples were analyzed with CST and results (positive signals for every pathogen/control) were visually detected by eye. As a reference quantitative polymerase chain reaction (qPCR) was performed. RESULTS The detection limit of CST revealed 1.2 × 104 for Treponema denticola (T.d.) and Tannerella forsythia (T.f.), 2.5 × 104 for Porphyromonas gingivalis (P.g.), 5.3 × 103 for Prevotella intermedia (P.i.), and 5.8 × 104 for Aggregatibacter actinomycetemcomitans (A.a.). Based on this maximum potential of positive detections, the sensitivities of CST in reference to qPCR were: T.d. (91.3%); T.f. (86.3%); P.g. (83.8%); P.i. (85.7%), and A.a. (100%). In regard to the clinical diagnosis, the CST assay and the qPCR method reached a sensitivity of 87.82% and 94%, respectively. The specificity for both methods was 100%. CONCLUSION This newly developed CST can detect five typical periodontal pathogens with a somewhat lower sensitivity towards qPCR that can be classified as "good.

Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms--which stain is suitable?

BACKGROUND There is confusion over the definition of the term "viability state(s)" of microorganisms. "Viability staining" or "vital staining techniques" are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data. DISCUSSION Many terms describe "vitality states" of microorganisms, however, several of them are misleading. Authors define "viable" as "capable to grow". Accordingly, staining methods are substitutes, since no staining can prove viability.The reliability of a commercial "viability" staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the "viability" kit are dependent on the stains' concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique.To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research.Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported. SUMMARY - The nomenclature regarding "viability" and "vitality" should be used carefully.- The manual of the commercial "viability" kit itself points out that the kit is not suitable for natural multispecies biofilm research, as supported by an array of literature.- Results obtained with various stains are influenced by the relationship between bacterial counts and the amount of stain used in the test. Corresponding vitality data are prone to artificial shifting.- As microbiological parameter the Plating Efficiency should be used for comparison.- Ethidium Bromide is mutagenic. Researchers should be aware that alternative staining compounds may also be or even are mutagenic

Clinical Evaluation of a New Electronic Periodontal Probe: A Randomized Controlled Clinical Trial

Precise measurements of periodontal parameters (such as pocket depths: PPD, gingival margins: GM) are important for diagnosis of periodontal disease and its treatment. Most examiners use manual millimeter-scaled probes, dependent on adequate pressure and correct readouts. Electronic probes aim to objectify and facilitate the diagnostic process. This randomized controlled trial compared measurements of a standard manual (MP) with those of an electronic pressure-sensitive periodontal probe (EP) and its influence on patients’ acceptance and practicability. In 20 patients (2436 measuring points) PPD and GM were measured either with MP or EP by professionals with different levels of experience: dentist (10 patients), 7th and 10th semester dental students (5 patients each). Time needed was measured in minutes and patients’ subjective pain was evaluated by visual analogue scale. Differences were analyzed using the generalized estimating equations approach (GEE) and paired Wilcoxon tests. Mean PPD varied with ΔPPD 0.38 mm between both probes, which was significant (p < 0.001), but GM did not (ΔREC 0.07 mm, p = 0.197). There was a statistically significant correlation of both probes (Spearman’s rho correlation coefficient GM: 0.674, PPD: 0.685). Differences can be considered robust (no deviation in either direction). The comparison of time needed and pain sensitivity did not result in statistically significant differences (p > 0.05)

Antibacterial Effect and Substantivity of Toothpaste Slurries In Vivo.

PURPOSE This double-blind, clinical, cross-over study evaluated the antibacterial effect of three toothpastes (ASF, HTP and STP) and a chlorhexidine mouthrinse (0.2%; CHX; positive control) after a single application on established biofilm over a period of 24 h (substantivity). MATERIALS AND METHODS Twenty-four subjects refrained from all oral hygiene measures for a period of 72 h. After 48 h, a baseline biofilm sample was taken and vitality of the biofilm flora was examined (baseline, VF0). Then they rinsed for 1 min with one of the randomly allocated, freshly prepared toothpaste slurries (ASF, HTP, STP) or CHX. Further biofilm samples were taken every second hour up to 14 h as well as 24 h after rinsing, and biofilm vitality was assessed (VF2-24). After a wash-out period of 4 days, a new test cycle was started. RESULTS All subjects (18 female, 6 male) finished the four test cycles. At VF2, all products showed a statistically significant reduction in vitality compared to VF0 (p<0.05). CHX and ASF revealed the most pronounced effect (49% and 40% reduction), while the other toothpastes (HTP: 24%, STP: 11%) reached lower but still statistically significant effects. At each further time point CHX and ASF showed the lowest biofilm vitality. ASF demonstrated a significant antibacterial effect on dental biofilm over a 24-h period compared to baseline and superiority over both other toothpastes at time points VF2-VF14. CONCLUSION ASF toothpaste showed a significant antibacterial action on biofilm and a high substantivity which was maintained up to 24 hours

Glucose-6-Phosphatase-Dehydrogenase activity as modulative association between Parkinson's disease and periodontitis.

UNLABELLED The association between periodontitis (PD) and Parkinson's disease (PK) is discussed due to the inflammatory component of neurodegenerative processes. PK severity and affected areas were determined using the following neuropsychological tests: Unified Parkinson's Disease Rating Score (UPDRS) and Hoehn and Yahr; non-motoric symptoms by Non-Motor Symptoms Scale (NMSS), and cognitive involvement by Mini-Mental State Examination (MMSE). Neuroinflammation and the resulting Glucose-6-Phosphatase-Dehydrogenase (G6PD) dysfunction are part of the pathophysiology of PK. This study aimed to evaluate these associations in periodontal inflammation. Clinical data and saliva-, serum-, and RNA-biobank samples of 50 well-characterized diametric patients with PK and five age- and sex-matched neurologically healthy participants were analyzed for G6PD function, periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Campylobacter rectus, Fusobacterium nucleatum, and Filifactor alocis), monocyte chemoattractant protein (MCP) 1, and interleukin (IL) 1-beta. Regression analysis was used to identify associations between clinical and behavioral data, and t-tests were used to compare health and disease. Compared with PK, no pathogens and lower inflammatory markers (p < 0.001) were detectible in healthy saliva and serum, PK-severity/UPDRS interrelated with the occurrence of Prevotella intermedia in serum as well as IL1-beta levels in serum and saliva (p = 0.006, 0.019, 0.034), Hoehn and Yahr correlated with Porphyromonas gingivalis, Prevotella intermedia, RNA IL1-beta regulation, serum, and saliva IL1-beta levels, with p-values of 0.038, 0.011, 0.008, <0.001, and 0.010, while MMSE was associated with Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, serum MCP 1 levels, RNA IL1-beta regulation and G6PD serum activity (p = 0.036, 0.003, 0.045, <0.001, and 0.021). Cognitive and motor skills seem to be important as representative tests are associated with periodontal pathogens and oral/general inflammation, wherein G6PD-saliva dysfunction might be involved. CLINICAL TRIAL REGISTRATION https://www.bfarm.de/DE/Das-BfArM/Aufgaben/Deutsches-Register-Klinischer-Studien/_node.html, identifier DRKS00005388

Nonsurgical treatment of aggressive periodontitis with photodynamic therapy or systemic antibiotics. Three-month results of a randomized, prospective, controlled clinical study

The aim of this randomized, controlled clinical study was to compare the short-term effects of nonsurgical periodontal therapy with the additional administration of systemic antibiotics (AB) and the same therapy with additional photodynamic therapy (PDT) in the treatment of patients with aggressive periodontitis (AP). Thirty-six patients with AP received full-mouth nonsurgical periodontal treatment (SRP) and were then randomly divided into two groups of 18 subjects each. Group AB received amoxicillin and metronidazole three times a day for 7 days. Group PDT received two applications of PDT on the day of SRP as well as at follow-up after 7 days. The following clinical parameters were measured at baseline and 3 months after therapy: plaque index (PLI), bleeding on probing (BOP), probing depth (PD), gingival recession (GR), and clinical attachment level (CAL). After 3 months, PD was significantly reduced in both groups (from 5.0±0.8 mm to 3.2±0.4 mm with AB, and 5.1±0.5 mm to 4.0±0.8 mm with PDT; both p<0.001), while AB revealed significantly lower values compared to PDT (p = 0.001). In both groups, GR was not significantly changed. CAL was significantly reduced in both groups (PDT: 5.7±0.8 mm to 4.7±1.1 mm; p=0.011; AB: 5.5±1.1 mm to 3.9±1.0 mm; p<0.001) and differed significantly between the groups (p=0.025). The number of residual pockets (PD ≥4 mm) and positive BOP was reduced by AB from 961 to 377, and by PDT from 628 to 394. Pockets with PD ≥7 mm were reduced by AB from 141 to 7, and by PDT from 137 to 61. After 3 months, both treatments led to statistically significant clinical improvements. The systemic administration of antibiotics, however, resulted in significantly higher reduction of PD and a lower number of deep pockets compared to PDT

Table_1_Glucose-6-Phosphatase-Dehydrogenase activity as modulative association between Parkinson’s disease and periodontitis.pdf

The association between periodontitis (PD) and Parkinson’s disease (PK) is discussed due to the inflammatory component of neurodegenerative processes. PK severity and affected areas were determined using the following neuropsychological tests: Unified Parkinson’s Disease Rating Score (UPDRS) and Hoehn and Yahr; non-motoric symptoms by Non-Motor Symptoms Scale (NMSS), and cognitive involvement by Mini-Mental State Examination (MMSE). Neuroinflammation and the resulting Glucose-6-Phosphatase-Dehydrogenase (G6PD) dysfunction are part of the pathophysiology of PK. This study aimed to evaluate these associations in periodontal inflammation. Clinical data and saliva-, serum-, and RNA-biobank samples of 50 well-characterized diametric patients with PK and five age- and sex-matched neurologically healthy participants were analyzed for G6PD function, periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Campylobacter rectus, Fusobacterium nucleatum, and Filifactor alocis), monocyte chemoattractant protein (MCP) 1, and interleukin (IL) 1-beta. Regression analysis was used to identify associations between clinical and behavioral data, and t-tests were used to compare health and disease. Compared with PK, no pathogens and lower inflammatory markers (p Clinical trial registrationhttps://www.bfarm.de/DE/Das-BfArM/Aufgaben/Deutsches-Register-Klinischer-Studien/_node.html, identifier DRKS00005388.</p