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Objectives To determine if minimally invasive non-surgical therapy (MINST) outperforms classical non-surgical periodontal therapy for stage III periodontitis with primarily suprabony (horizontal) type defects. Materials and methods In a split-mouth randomised controlled trial, 20 patients’ dental quadrants were randomly assigned to MINST or classical non-surgical treatment. The primary outcome variable was the number of sites with probing pocket depth ≥ 5 mm and BOP. Treatment method, tooth type, smoking status, and gender were evaluated using a multivariate multilevel logistic regression model. Results After 6 months, the percentage of sites with PD ≥ 5 mm and BOP that healed (MINST = 75.5%; control group = 74.1%; p = 0.98), and the median number of persisting sites (MINST: 6.5, control group: 7.0; p = 0.925) were similar in both groups. In the test and control groups, respectively, median probing pocket depths (2.0 mm vs. 2.1 mm) and clinical attachment level (1.7 mm vs. 2.0 mm) changed significantly (p p = 0.037). Men (OR = 0.52, p = 0.014) and non-molars (OR = 3.84, p 0.001) had altered odds for healing of sites with PD ≥ 5 mm and BOP. Conclusions MINST reduces gingival recession associated with molar teeth, although it performs similarly to traditional non-surgical therapy in treating stage III periodontitis with predominately horizontal-type defects. Clinical relevance MINST performs similarly to non-surgical periodontal therapy in stage III periodontitis with predominantly suprabony defects. Trial registration Clinicaltrials.gov (NCT04036513) on June 29, 2019.

Current discoveries imply a connection between periodontitis and metabolic associated fatty liver disease (MAFLD). This study aimed to determine the prevalence of periodontitis and MAFLD in obese patients with BMI40, employing the most reliable diagnostic methods, namely liver biopsy, and detailed periodontal examination.Liver biopsy and periodontal examination were performed in 30 obese patients with BMI BMI40 undergoing bariatric surgery. Kleiner's classification was used to determine non-alcoholic steatohepatitis (NAS) activity score, non-alcoholic steatohepatitis (NASH) and liver fibrosis. The periodontal condition was classified following the recent AAP/EFP classification. Patients were divided into periodontitis (PG) and non-periodontitis groups (NPG). Data on systemic health parameters were collected from patients' medical records. Descriptive statistics and simple statistical tests were used to determine the differences between the two groups.The prevalence of NASH in the sample was 43% (13/30), borderline NASH 37% (11/30), while fibrosis stage 1 was most common (72%, [22/30]). Periodontitis prevalence was 67% (20/30), while all non-periodontitis patients (33%; 10/30) exhibited gingivitis. PG and NPG did not differ in NAS or NASH prevalence (p0.05). However, the periodontitis group showed higher C-reactive protein levels, while NPG showed higher gamma-glutamyl transpeptidase levels (p0.05).The study results suggest the considerable prevalence of MAFLD, periodontitis and gingivitis in obese patients with BMI40 undergoing bariatric surgery. Patients with periodontitis had higher CRP levels, while those with gingivitis presented higher gamma-glutamyl transpeptidase levels.

IntroductionWe aimed to determine if periodontal health deteriorates after bariatric surgery (BS).Material and methodsA search was performed in Medline and Embase, for prospective cohort studies with data on change in periodontal parameters after BS. Me�ta-analysis was performed with available data.ResultsThe results of 4 included studies consistently show significant (pConclusionsBS may have a transient negative consequence on periodontal health.

Purpose: The aim of this study was to evaluate the survival and success rates of oral rehabilitations in a shortened maxillary dental arch and expanded maxillary sinus with 4-mm extrashort implants splinted to longer ones. Materials and methods: One 10-mm and one/two extrashort 4-mm titanium-zirconium SLActive tissue-level dental implants were inserted into 11 patients with limited vertical bone availability due to an expanded maxillary sinus antrum. Immediately and 6 months after insertion, implant stability resonance frequency analysis (RFA) values were assessed with an Osstell device. Splinted crowns combining 4- and 10-mm implants were supplied to all 11 cases. Results: In 10 cases, the bone quality was type III, and in one case, type IV. Among 17 4-mm and 11 10-mm implants, the median RFA values were 61 (interquartile ranges [IQR]: 59 to 64) and 66 (IQR: 64 to 72). One 4-mm implant failed to osseointegrate and was removed. After 6 months of healing, secondary-stability measurements of 16 of the remaining 4-mm implants increased to 68 (IQR: 62 to 72) and of 10-mm implants to 78 (IQR: 77 to 80). After 1 year, all (11/11) oral rehabilitations supported by 10-mm (11/11) and 4-mm (16/16) implants were functional. The medians and IQRs of the probing depths (median: 2.8 mm, IQR: 2.3 to 3.1 mm vs median: 2.9 mm, IQR: 2.4 to 3.1 mm) and the crestal bone loss (median: 0.75 mm, IQR: 0 to 0.9 mm vs median: 0.22 mm, IQR: 0 to 0.4 mm) for the 10-mm and 4-mm implants, respectively, were similar. Conclusion: Rehabilitations with splinted crowns combining 4- and 10-mm implants demonstrated a favorable 1-year performance in a shortened maxillary dental arch.

This study aimed to rehabilitate shortened maxillary dental arch with splinted crowns by connecting ultra-short implants with longer ones. In the posterior maxilla of 11 patients, one 10-mm (n = 11) and one or two ultra-short 4-mm (n = 17) dental implants were inserted. The insertion torque was lower than 20 Ncm in 55% of the 10-mm implants and in 94% of the 4-mm implants (P > .05). Median (range) implant stability quotients at the time of insertion and after 6 months were 61 (14 to 72) and 68 (51 to 79), respectively, for 4-mm implants, and 66 (52 to 78) and 78 (60 to 83), respectively, for 10-mm implants (P .05). Splinted crowns combining 4- and 10-mm implants may contribute to a better force distribution in the treatment with ultra-short implant-supported prosthesis in the posterior maxilla.

To determine if periodontitis patients benefit from treatment with Lactobacillus brevis and Lactobacillus plantarum strains, applied into periodontal pockets as gel and thereafter taken as lozenges, as an adjunct to scaling and root planing (SRP).In a double-blind, randomized, placebo-controlled trial, 40 patients received scaling and root planing (SRP) in two sessions within 7 days. Patients then received either probiotic gel and lozenges (n = 20) or placebo (n = 20). The primary outcome variable was the number of diseased sites (DS: PD 4 mm + BOP) at the 3-month re-evaluation. The effects of gender, age, probiotic therapy, presence of Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans, smoking, tooth being a molar and interdental location were evaluated using a multivariate multilevel logistic regression model.The number of DS after 3 months was similar in the test (Me = 8, IQR = 5-11) and control (Me = 5, IQR = 1-10) groups. Both groups showed substantial but equivalent improvements in periodontal parameters. The logistic regression showed higher odds for the healing of gingival bleeding (OR = 2.12, p = 0.048) and lower odds for the healing of DS (OR = 0.51; p 0.001) in the probiotic group.Patients with periodontitis benefit from adjunctive use of probiotics containing L. brevis and L. plantarum in terms of reduction of gingival bleeding. However, adjunctive probiotics increase the number of persisting diseased sites with PD 4 mm and BOP.The adjunctive use of probiotics containing L. brevis and L. plantarum strains in treating chronic periodontitis results in a higher number of residual diseased sites when compared with SRP + placebo; its use is therefore unfounded.

Objective Current approaches for soft tissue thickness evaluation and visualization still represent a challenge for full extent evaluation and visualization. The aim of this clinical technique article is to introduce a novel approach for comprehensive visualization and precise evaluation of oral soft tissue thickness utilizing a fusion of optical 3D and cone-beam computed tomography (CBCT) images. Clinical considerations 3D models of the maxilla were obtained by CBCT imaging and intraoral scanning. The CBCT images were reconstructed to standard tessellation language (STL) file format models by segmentation of teeth and bone using implants planning software. 3D soft tissues and teeth models were obtained by intraoral scanning and were exported in STL file format as well. 3D multimodal models were then superimposed using best-fit matching on teeth. Soft tissue thickness was then visualized and evaluated with a 3D color-coded thickness map of gingival and palatal areas created by surface comparison of both 3D models. Additionally, threshold color-coding was used to increase comprehensibility. Palatal areas were further visualized and evaluated for the optimal donor site. Conclusions A novel approach for 3D evaluation and visualization of masticatory mucosa thickness presents all available 3D data in a comprehensible, "clinician-friendly" manner, using threshold regions and clinically relevant views. Clinical significance Proposed approach could provide comprehensive presurgical treatment planning in periodontal plastic surgery and implantology without additional invasive procedures for the patient, resulting in more predictable treatment, improved outcomes, and reduced risk for complications.

Instability of the surgical guide is an overlooked factor that can result in a difference between the planned and the actual positions of an implant. Our aim was to compare the stability of the retentive surgical guide (RSG) with a conventional surgical guide (CSG) in an in-vitro experiment.A platform to evaluate the stability of the surgical guide was designed using 3D-modelling software (Meshmixer 3.5, Autodesk). Imaging data from 15 patients with a single missing tooth were used to plan the virtual implant. Two surgical guides were designed (Blue Sky Plan 4.8, Blue Sky Bio) and 3D printed (Form2, Dental SG resin, Formlabs) for each case: the CSG with the default, predetermined software settings, and the RSG, designed on a dental model with a 0.1-mm undercut and altered production parameters (reduced guide-to-teeth offset of 0.07 mm, reduced guide thickness of 2.3 mm and a retentive clasp in a marginal area). The dental models were reproducibly secured on the testing platform using a digital force gauge, and the surgical guides were positioned. An increasing force of 0.1 N, 1 N, 2.5 N, and 5 N was sequentially applied from the buccal and the oral directions to the surgical guide via a drill handle. For each force, either the magnitude of the guide's displacement was captured with an intra-oral scanner (CEREC Omnicam AC, Dentsply Sirona; software version: SW 4.5.2) or the dislodgement of the guide was recorded. Scans were imported for analysis (GOM Inspect 2018, GOM GmbH), and library files of the surgical guides and implants were superimposed as a joined complex. The deviation of the implant's position was calculated from the displacement of the guide's position RESULTS: Three-way repeated measures using ANOVA revealed a more significant guide displacement and virtually projected implant deviation in the CSG group than the RSG group and with increasing force in all the deviation parameters. Both groups showed greater resistance to the displacement with the force applied from the oral direction than the buccal direction. The application of the force in the buccal direction resulted in guide dislodgements of 13% and 0% for the CSG and RSG, respectively. In the oral direction, the dislodgement rates were 33% and 7% for the CSG and RSG, respectively.Within the limitations of this study, the retentive design increased the stability of the surgical guide and, consequently, the accuracy of the virtually projected implants in comparison to the conventional surgical guide designed using the default settings. Clinical trials are needed to confirm its advantages in clinical use.With a simple modification to the design, the surgical guide retention provided greater stability, with smaller deviations under loading; this resulted in improved implant precision parameters without requiring additional materials or software. Further studies are needed to assess the clinical feasibility of this surgical guide with improved retention and function.

The present study aimed to compare the conventional clinical and a digital method for evaluating differences in gingival recession (ΔREC) in patients with advanced periodontitis treated with the non-surgical treatment protocol.Agreement between the methods was evaluated on a sample of ten patients with periodontitis (stage III/IV, grade B/C) with acquired clinical measurements and digital models from baseline (T0) and 12-months after non-surgical treatment of periodontitis (T1). The evaluation was performed on maxillary teeth from right to left second premolar resulting in overall 99 teeth. Clinical evaluation was performed by subtracting the distance measurements between gingival margin and cemento-enamel junction, obtained at T0 and T1 by a calibrated examiner (intra-examiner agreement90%). The digital evaluation was performed directly by measuring the distance between the gingival margins on superimposed T0 and T1 digital models. Using Bland-Altman and statistical analysis, all six measurements sites around each included tooth (n=594) acquired with both methods were compared.Median ΔREC (5th and 95th percentile) acquired with a conventional clinical and digital method was 0.0mm (-2.0 - 1.0) and -0.4mm (-1.6 - 0.8), respectively (p0.0001). The complete agreement between rounded digital and clinical ΔREC values was only 38%, revealing high disagreement also confirmed by Bland-Altman analysis with 95% limits of agreement ranging from -2.6 to 1.8mm. Absolute differences between the methods higher than 0.5 and 1 mm, was found in 61% and 38% of measurement sites, respectively.The conventional clinical method for ΔREC evaluation exhibits lower sensitivity and accuracy than the digital method.The quality of both clinical and research data in periodontology and implantology can be considerably improved by the digital method while still preserving the compatibility with the conventional clinical method.

Replantation and retention of ankylosed tooth after pubertal growth spurt enables stepwise replacement of both central incisors with implants. Partial extraction contributes to natural gingival contour., Replantation and retention of ankylosed tooth after pubertal growth spurt enable stepwise replacement of both central incisors with implants. Partial extraction contributes to natural gingival contour.

Background: Bariatric surgery (BS) patients may experience the progression of periodontitis during recovery. We aimed to determine whether non-surgical periodontal therapy before BS improves the periodontal and systemic health parameters after the surgery. Methods: BS candidates with periodontitis were randomized into the test (TG) and control group (CG). One month before BS (pre-BS), patients in the TG (n = 15) received non-surgical periodontal therapy, while patients in the CG (n = 15) received only mechanical plaque removal. Patients were re-examined 3 and 6 months after BS. Differences between the TG and CG in clinical periodontal parameters, systemic health-related serum biomarkers, parameters of obesity, and prevalence of obesity-related diseases were evaluated. Results: From the 30 included patients, 26 were re-examined at 3 months and 20 patients at 6 months. Periodontal parameters bleeding on probing (p = 0.015), periodontal pocket dept (PPD, p = 0.0015), % PPD > 4 mm (p < 0.001), and full-mouth plaque levels (p = 0.002) were lower in the TG than in the CG at 6 months after BS. There is a general improvement in systemic health after BS without significant differences (p > 0.05) between the TG and CG at the 6-month follow-up. The TG shows a tendency for improvement in metabolic syndrome components at the 6-month follow-up compared to pre-BS (p < 0.05). Conclusions: Non-surgical periodontal therapy in periodontitis patients before the BS may improve periodontal health 3 and 6 months after the surgery. The possible benefits of periodontal therapy on the overall health of BS patients should be further explored.

Background It is well recognized that dental procedures represent a potential way of infection transmission. With the COVID‐19 pandemic, the focus of dental procedure associated transmission has rapidly changed from bacteria to viruses. The aim was to develop an experimental setup for testing the spread of viruses by ultrasonic scaler (USS) generated dental spray and evaluate its mitigation by antiviral coolants. Methods In a virus transmission tunnel, the dental spray was generated by USS with saline coolant and suspension of Equine Arteritis Virus (EAV) delivered to the USS tip. Virus transmission by settled droplets was evaluated with adherent RK13 cell lines culture monolayer. The suspended droplets were collected by a cyclone aero‐sampler. Antiviral activity of 0.25% NaOCl and electrolyzed oxidizing water (EOW) was tested using a suspension test. Antiviral agents' transmission prevention ability was evaluated by using them as a coolant. Results In the suspension test with 0.25% NaOCl or EOW, the TCID50/ml was below the detection limit after 5s. With saline coolant, the EAV‐induced cytopathic effect on RK13 cells was found up to the distance of 45 cm, with the number of infected cells decreasing with distance. By aero‐sampler, viral particles were detected in concentration ≤4.2 TCID50/ml. With both antiviral agents used as coolants, no EAV‐associated RK‐13 cell infection was found. Conclusion We managed to predictably demonstrate EAV spread by droplets due to USS action. More importantly, we managed to mitigate the spread by a simple substitution of the USS coolant with NaOCl or EOW. This article is protected by copyright. All rights reserved

During an ultrasonic scaler (USS) operation, droplets and aerosol are generated that may contribute to the transmission of viruses contained in saliva and gingival crevicular fluid. The purpose of this research was to develop an experimental model for testing the spread of viruses during USS instrumentation and to examining the prevention of spreading by replacing the coolant with an antiviral agent. In a virus transmission tunnel, USS operation with saline coolant and delivery of a viral suspension to the vicinity of USS tip generated droplets and aerosol containing Equine Arteritis Virus (EAV). Evaluation of droplet transmission was evaluated with adherent 48h cell culture monolayer RK13 cell lines in standard 48-well-plates positioned at a distance from 30 to 55 cm. The aerosol was collected by a cyclone aero-sampler flow of 100l/min. Antiviral activity of 0.25% sodium hypochlorite or electrolyzed water (EOW) was tested by suspension test. The two tested antiviral agents’ transmission prevention ability was evaluated by repeating the same experiment as with saline coolant. All experiments were repeated twice. With saline coolant, the cytopathic effect on cells was found in cells up to the distance of 45 cm, with the number of infected wells decreasing with distance. Viral particles were detected in only one AS in a very low concentration (≤4.2 TCID50/ml). In suspension test of 0.25% NaOCl and EOW, the TCID50/ml was below detection limit after 5s. With both antiviral agents, no cytopathic effect was found. However, the cytotoxic effect of 0.25% NaOCl was evident up to the distance of 35 cm. By USS activity, EAV could be transmitted by droplets up to a distance of 45 cm. Both antiviral agents could prevent virus droplet transmission. The transmission of EAV by aerosol yielded inconclusive results.

To investigate the subgingival microbiological profiles of patients with periodontitis, to determine their stage and grade scores and to evaluate the differences in the microbiota among different stages and grades. Materials and Methods: Sixty-seven (n = 67) periodontitis patients were selected. Periodontitis staging and grading, following the 2018 classification system, were defined. Following a clinical examination, subgingival samples were taken from the deepest periodontal pocket of each quadrant for cultivation, identification and quantification. The prevalence, proportion and counts of nine selected periodontal pathogens were determined, and differences between periodontitis stages III and IV and grades B and C were assessed. Results: All nine cultivable periodontal bacteria were detected, of which the most prevalent was P. intermedia (91.0%) and the least prevalent were E. corrodens (9.0%) and C. ochracea (9.0%). The frequency of detection of the two main target pathogens, A. actinomycetemcomitans and P. gingivalis, was 41.8% and 76.1%, respectively. The prevalence (grade B: 80.6%, grade C: 55.6%, p = 0.035) and total counts (grade B: 19.8 colony forming units - CFU/ml-4 (1.9-52.8); grade C: 4.0 CFU/ml-4 (0.0-26.4); p = 0.022) of F. nucleatum were statistically significantly higher in grade B than in grade C periodontitis patients, whereas the counts of P. gingivalis and A. actinomycetemcomitans were similar between grades and stages. Conclusion: Our study suggests that relevant differences between the various grades of periodontitis exist only in the numbers of F. nucleatum. Prevalence and quantities of other cultivable species between different stages and grades of periodontitis seem to be similar.

Background Our aim was to determine if azithromycin therapy, as an adjunct to scaling and root planing (SRP), decreases the number of pathobiontic subgingival plaque species and sites demonstrating pocket depth (PD) ≥ 5 mm and bleeding on probing (BOP) 6 months post-treatment. Methods In a double-blind randomized parallel-arm placebo-controlled trial, 40 patients received nonsurgical periodontal treatment in two sessions within 7 days. Patients then received systemic antibiotic therapy (n = 20, azithromycin 500 mg/day for 3 days) or placebo (n = 20). Pooled microbiologic samples were taken before and 6 months after therapy and analysed by established culture methods. The primary outcome variable was the number of sites with PD ≥ 5 mm and BOP at the 6-month re-evaluation. Using multivariate multilevel logistic regression, the effects of gender, age, antibiotic therapy, presence of P. gingivalis or A. actinomycetemcomitans, smoking, tooth being a molar and interdental location were evaluated. Results The number of sites with PD ≥ 5 mm and BOP after 6 months was similar in the test (Me = 4, IQR = 0–11) and control (Me = 5, IQR = 1–22) group. Adjunctive azithromycin treatment, compared to SRP alone, resulted in more frequent eradication of A. actinomycetemcomitans (p = 0.013) and C. rectus (p = 0.029), decreased proportion (p = 0.006) and total counts (p = 0.003) of P. gingivalis, and decreased proportion of C. rectus (p = 0.012). Both groups showed substantial but equivalent improvements in periodontal parameters, with no intergroups differences at initially shallow or deep sites. The logistic regression showed a lower odds ratio for healing of diseased sites on molars (OR = 0.51; p Conclusion Despite significant changes in numbers of A. actinomycetemcomitans, P. gingivalis and C. rectus, patients with periodontitis do not benefit from adjunctive systemic azithromycin in terms of number of persisting sites with PD ≥ 5 mm and BOP. Trial registration EUDRA-CT: 2015–004306-42; https://www.clinicaltrialsregister.eu/ctr-search/trial/2015-004306-42/SI, registered 17. 12. 2015.

Objectives The aim of the present review was to find, compare, and critically discuss digital methods for quantitative evaluation of gingival recessions dimensions. Data Collection of articles and classification related to digital evaluation of gingival recessions. Sources A search of PubMed, Web of Science, Scopus, and reference lists of articles was conducted up to April 2021 Study selection Twenty-two articles used digital evaluation of gingival recessions dimensions. The methods in the included articles were extracted, compared, and categorized. Results Digital measurements were performed on 2D intraoral photographs, 3D models, or cross-sections obtained from 3D models. Baseline measurement were performed for diagnostic and treatment planning and categorised into distance and area measurements. Follow-up evaluation of treatment was based either on repeating the “baseline” measurements and calculating differences or measuring differences directly on composite images, composed from two superimposed images obtained at two time-points. Direct measurements were categorised into distance, area, and volume measurements. Conclusions Digital evaluation predominantly means just digitalization of the established evaluation methods; therefore, increasing measurements accuracy and maintaining comparability with past studies. At present, a large variability of digital evaluation workflow among the included studies renders the comparison among different studies difficult if not impossible. The potential of digital evaluation seems not to have been fully exploited as only a few novel measurements and parameters introduced, i.e., volumetric evaluation of soft tissue dynamics. For reproducible and comparable studies in the future, the research should be aimed at evaluation, optimization and standardization of all phases of the digital evaluation. Clinical Significance Digital evaluation, based on 3D image superimposition is a promising approach as it increases measurements accuracy, maintains compatibility with past studies and simultaneously introduces novel evaluation possibilities.

Collagen matrices (CMs) could be used instead of connective tissue grafts (CTGs) for treatment of gingival recession (GR). The authors aimed to compare clinical outcomes after treatment of isolated GR affecting both maxillary canines of 10 patients (n = 10) with CM (OsteoBiol Derma) or CTG in conjunction with coronally advanced flap. After 12 months, CM and CTG showed complete correction in 7/10 and 10/10 of sites, respectfully, and percentage of root coverage was 85% ± 24% and 100%, respectively. For treatment of isolated GR, CM is a viable alternative to CTG.

Tongue piercing (TP) has been gaining in popularity, mainly among adolescents and young adults, and there has been a corresponding increase in reports of dental injury and adverse effects on periodontal tissue due to TP jewellery. The purpose of this study was to assess the type and prevalence of dental injury and periodontal complications in a group of Slovenian adolescents and young adults with TP in comparison with a control group.This case-control study included 17 subjects with TP (study group) and 28 subjects without TP (control group). Subjects were clinically examined for dental injuries, as classified according to Andreasen. Periodontal complications were evaluated using an estimation of clinical attachment loss (CAL) and gingival recession (GR). Differences in categorical variables were analyzed using the chi-squared or Fisher's exact test as appropriate. Differences between the groups in numerical variables were analyzed with the Mann-Whitney U-test. A multiple logistic regression model was built to test the association between dental injuries and TP when controlling for cofounders, namely age and gender of study participants. The significance level was set at α = 0.05 (two-tailed).Subjects with TP had 12.2 higher odds of dental injury (CI = 2.2-67.7, p = 0.004) compared with subjects without TP. Enamel fractures were more prevalent in the study group (p = 0.002). Frequencies of other dental injuries in the two groups were similar. GR was significantly associated with TP (p = 0.008), while differences in CAL between the groups were not statistically significant.Subjects with tongue piercing had more dental injuries and gingival recession.

The periodontopathogen Aggregatibacter actinomycetemcomitans synthesizes several virulence factors, including cytolethal distending toxin (CDT). The active CDT holoenzyme is an AB-type tripartite genotoxin that affects eukaryotic cells. Subunits CdtA and CdtC (B-components) allow binding and intracellular translocation of the active CdtB (A-component), which elicits nuclear DNA damage. Different strains of A. actinomycetemcomitans have diverse virulence genotypes, which results in varied pathogenic potential and disease progression. Here, we identified an A. actinomycetemcomitans strain isolated from two patients with advance chronic periodontitis that has a regular cdtABC operon, which, however, codes for a unique, shorter, variant of the CdtB subunit. We describe the characteristics of this CdtBΔ116–188, which lacks the intact nuclear localisation signal and the catalytic histidine 160. We show that the A. actinomycetemcomitans DO15 isolate secretes CdtBΔ116–188, and that this subunit cannot form a holotoxin and is also not genotoxic if expressed ectopically in HeLa cells. Furthermore, the A. actinomycetemcomitans DO15 isolate is not toxic, nor does it induce cellular distention upon infection of co-cultivated HeLa cells. Biological significance of this deletion in the cdtB remains to be explained.

Aim The aim was to assess the potential trends in Periodontology and Periodontics in Europe that might be anticipated by the year 2025, using the Delphi method. Material and Methods The expert opinion of 120 experts was sought through the use of an open-ended questionnaire, developed by an advisory group, containing 40 questions concerning the various trends in periodontology. Results The experts (113 responders) expect a stabilization of the prevalence of periodontitis, both for the chronic as well as the aggressive cases, but an increase in implant-related diseases up to the year 2025. Concurrently, the importance of implants is seen to be increasing. They foresee an increased demand for postgraduate periodontology and implantology training. This is mirrored in an increase in publications for implant dentistry and increase in demand and need for training. Concerning the patients, better-informed individuals seeking more routine check-ups are expected. Conclusion A continued need for specialized periodontists, but also well trained dental practitioners is foreseen for next decade in Europe. Apart from periodontology they will be increasingly exposed to and trained in implant dentistry.

Objectives Expression of neuronal neuropeptides in inflammatory conditions is altered. The changes in expression of substance P (SP) and calcitonin gene-related peptide (CGRP) in ipsilateral and contralateral trigeminal ganglion (TG) neurons were investigated by immunohistochemistry one week after unilateral ligature-induced periodontitis in rats. Design A retrograde nerve tracer Fluorogold (FG) was applied into the gingival sulcus of the second maxillary molar to identify the neurons in TG that specifically innervate the inflamed gingivomucosa. In addition, neurons from the corresponding maxillary and the adjacent mandibular–ophthalmic regions in TG were analysed. Results Statistically significantly higher frequencies of CGRP-positive neurons, regardless of their size, were found in TG ipsilateral to the periodontitis (83% and 73% in FG-labelled and maxillary regions, respectively) than in the control group without periodontitis (52% and 42% in FG-labelled and maxillary regions, respectively). The frequency of small FG-labelled SP-positive neurons in the ipsilateral TG (60%) was significantly higher than in the control TG (25%). In the contralateral TG the frequency of CGRP-positive neurons in maxillary region (66%) was significantly higher than in the control group. Surprisingly, the number of SP-positive neurons in all regions of contralateral TG decreased when compared to control and ipsilateral TGs. Conclusions Taken together, these results implicate a role of neurogenic component in the pathogenesis of periodontitis. The contralateral response in the TG could be mediated through the transmedian neurological pathways crossing in the trigeminal nuclear complex or through the systemic inflammatory reaction and the activation of the so called “neuro-immune axis”.

Proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) was found in inflamed periodontal tissues and many studies pointed to its significant role in development of periodontal disease. In this study, the influence of subcutaneously administered recombinant human TNF-alpha (rhTNF-alpha) on inflammatory reaction and periodontal breakdown in rats was analyzed during experimental periodontitis, induced by placing silk ligatures around the maxillary right second molar tooth. The rats were divided into two groups with five animals in each; the first group was infused subcutaneously with rhTNF-alpha via osmotic pumps for 2 weeks and the second group was infused with phosphate-buffered saline (PBS) in the same manner. Inflammatory reaction and periodontal breakdown was evaluated morphometrically on hematoxylin and eosin stained sections. Serum ionized calcium and inorganic phosphates were monitored colorimetrically. Serum calcium and phosphate levels were similar in rats receiving rhTNF-alpha and PBS. Ligation resulted in accelerated periodontal breakdown, while subcutaneous rhTNF-alpha administration by itself had no significant effect. Combined effect of subcutaneous rhTNF-alpha administration and ligation resulted in a significantly greater inflammatory reaction and periodontal breakdown then either treatment alone. We concluded that the subcutaneous administration of rhTNF-alpha accelerates the progression of experimental periodontitis in rats.

New findings regarding acetylcholinesterase (AChE) in the neuromuscular junction (NMJ), obtained in the last decade, are briefly reviewed. AChE is highly concentrated in the NMJs of vertebrates. Its location remains stable after denervation in mature rat muscles but not in early postnatal muscles. Agrin in the synaptic basal lamina is able to induce sarcolemmal differentiations accumulating AChE even in the absence of a nerve ending. Asymmetric A12 AChE form is the major molecular form of AChE in vertebrate NMJs. Extrajunctional suppression of this form is a developmental phenomenon. Motor nerve is able to reinduce expression of the A12 AChE form in the ectopic NMJs even in muscles with complete extrajunctional suppression of this form. The 'tail' of the A12 AChE form is made of collagen Q. It contains domains for binding AChE to basal lamina with ionic and covalent interactions. Muscle activity is required for normal AChE expression in muscles and its accumulation in the NMJs. In addition, the pattern of muscle activation also regulates AChE activity in the NMJs, demonstrating that the pattern of synaptic transmission is able to modulate one of the key synaptic components.

Background: In vitro studies demonstrated that proinflammatory cytokine tumor necrosis factor alpha (TNF-α) modulates bone metabolism. Objective: The aim of our study was to confirm the ability of TNF-α to induce osteoclastogenesis and bone resorption in an in vivo experiment, with the use of calvarial model in mice. Materials and methods: Twenty C57-Black mice were divided into four groups with five animals in each. The first group was infused subcutaneously on their back with recombinant mouse (rm) TNF-α via osmotic minipumps for 3 d, the second group was similarly infused with phosphate-buffered saline (PBS), the third group was infused with rmTNF-α to the region above the parietal bone and the fourth group with PBS in the same manner. Number of osteoclasts on parietal bone was determined morphometrically. Serum calcium and phosphates were monitored colorimetrically. Results: Serum calcium level and number of osteoclasts on parietal bone were significantly greater after infusion of rmTNF-α above the parietal bone, whereas after subcutaneous delivery these parameters were similar to the control group. Conclusion: We are concluding that TNF-α has the ability to change the bone metabolism in a paracrine manner only.

Periodontitis is characterized by periodontal tissue destruction, including the alveolar bone. One of its critical components is the release of pro-inflammatory neuropeptides from sensory nerve endings innervating the periodontium. Since nerve growth factor (NGF) has been reported to up-regulate neuropeptides in sensory neurons, we hypothesized that it would be increased in ligature-induced periodontitis in rats, and that systemic NGF neutralization would reduce the periodontitis-associated alveolar bone resorption. Real-time PCR analysis disclosed a statistically significant time-dependent up-regulation of NGF mRNA in gingiva during 2 weeks of periodontitis. Interestingly, NGF up-regulation was also detected in the contralateral gingiva. In addition, immunohistochemistry of trigeminal ganglion neurons innervating the gingivomucosa demonstrated increased expression of TrkA receptor for NGF. Systemic anti-NGF treatment during periodontitis significantly reduced interleukin-1β expression in gingiva and bilateral alveolar bone resorption. This suggests that NGF promotes periodontal inflammation and implicates a possible use of anti-NGF treatment in periodontitis.

The purpose of this study was to characterize and evaluate the expression of TrkA receptor in trigeminal ganglion (TG) neurons that innervate the rat gingivomucosal tissue. A retrograde nerve tracer Fluorogold (FG) was injected into the gingiva (group 1) or applied into the gingival sulcus (group 2) of the first right maxillary molar to identify the neurons in TG that innervate the gingivomucosa. After 10 days TG were dissected and FG fluorescence in neurons was observed under UV light microscope. To draw a comparison, approximately 1000 neurons per ganglion from the entire TG (group 3) and approximately 350 neurons per ganglion from the maxillary region in TG (group 4), were analyzed. Expression of TrkA receptor in TG neurons was investigated by immunohistochemistry. About 70% of neurons in groups 1 and 2 contained TrkA receptor, which was statistically significantly more than in groups 3 (41%) and 4 (38%). FG-labeled TrkA-immunopositive neurons were predominantly small or medium-sized (less than 1200microm(2)). However, the neurons innervating the rat gingivomucosa were on average larger than the neurons in the entire TG or in the maxillary region. In conclusion, the majority of neurons in TG that innervate the rat gingivomucosa are small or medium-sized, contain TrkA receptor and are most probably nociceptive.

Summary Objectives The purpose of this study was to identify and characterise the neuropeptide content and the size of trigeminal ganglion (TG) neurons innervating the rat gingivomucosal tissue. Design Retrograde nerve tracer Fluorogold (FG) was injected into the gingiva (group 1, n = 5) or applied into the gingival sulcus (group 2, n = 5) of the first right maxillary molar. After 10 days, the ganglia were dissected and FG fluorescence was observed under UV light microscope. Expression of calcitonin gene-related peptide (CGRP) and substance P (SP) in FG-labelled neurons was investigated by immunohistochemistry. Cross-sectional areas of neuron cell bodies containing FG were determined. As a control group, approximately 1000 neuron cell bodies representing the entire TG neuron population was evaluated in five trigeminal ganglia. Results In group 1, the percentages of neurons containing CGRP (median 63%, range 48–72%) and SP (median 64%, range 54–64%) were significantly greater than in the control group (CGRP: median 43%, range 42–47% and SP: median 23%, range 21–27%). In group 2, only the percentage of neurons containing SP (median 50%, range 40–56%) was significantly greater than in the control group. FG-labelled neurons were predominantly small or medium sized (less than 1200 μm 2 ). The neurons in the group 1 were significantly smaller than in group 2. In both experimental groups, immunopositive neurons were significantly smaller than immunonegative neurons. Conclusions The majority of neurons in TG that innervate the rat gingivomucosa are small or medium sized and contain CGRP and SP.

Expression of acetylcholine receptors (AChRs) in the extrajunctional muscle regions, but not in the neuromuscular junctions, is repressed by propagated electric activity in muscle fibers. During regeneration, subsynaptic-like specializations accumulating AChRs are induced in new myotubes by agrin attached to the synaptic basal lamina at the places of former motor endplates even in the absence of innervation. We examined whether AChRs still accumulated at these places when the regenerating muscles were ectopically innervated and the former synaptic places became extrajunctional. Rat soleus muscles were injured by bupivacaine and ischemia to produce complete myofiber degeneration. The soleus muscle nerve was permanently severed and the muscle was ectopically innervated by the peroneal nerve a few millimeters away from the former junctional region. After 4 weeks of regeneration, the muscles contracted upon nerve stimulation, showed little atrophy and the cross-section areas of their fibers were completely above the range in non-innervated regenerating muscles, indicating successful innervation. Subsynaptic-like specializations in the former junctional region still accumulated AChRs (and acetylcholinesterase) although no motor nerve endings were observed in their vicinity and the cross-section area of their fibers clearly demonstrated that they were ectopically innervated. We conclude that the expression of AChRs at the places of the former neuromuscular junctions in the ectopically innervated regenerated soleus muscles is activity-independent.