Evaluation of early bacterial adhesion on CAD/CAM dental materials: an in situ study.

The aim of this research was to determine if there are differences in early bacterial adhesion among CAD/CAM dental materials after 24 h exposure in the oral environment. One hundred twenty specimens were prepared according to the manufacturer's recommendations and divided into six groups: RBC (resin-based composite), PMMA (polymethyl methacrylate), PEEK (polyether ether ketone), ZP (zirconia polished), ZG (zirconia glazed), and cobalt-chromium alloy (CoCr alloy). Twenty healthy participants were instructed to carry an intraoral device with six specimens, one per group, for 24 h. Thereafter, real-time polymerase chain reaction (qPCR) and scanning electron microscopy (SEM) analyses enabled quantification and 2D view of biofilm formed on the specimens' surfaces. Kruskal-Wallis test and Dunn's post hoc analysis were used for inter-group comparison and data were presented as median (minimum-maximum). RBC specimens accumulated less bacteria, in comparison with ZG (p = 0.017) and PEEK specimens (p = 0.030), that dominated with the highest amount of adhered bacterial biofilm. PMMA, CoCr, and ZP specimens adhered more bacteria than RBC (p > 0.05), and less than ZG (p > 0.05) and PEEK (p > 0.05). The bacterial number varied considerably among participants. The obtained results enable a closer view into the susceptibility of CAD/CAM materials to microorganisms during the presence in the oral environment, which can be beneficial for a proper selection of these materials for a variety of dental restorations.

Periodontal health status in systemic sclerosis patients: Systematic review and meta-analysis.

OBJECTIVE: The present systematic review and meta-analysis aimed to evaluate the periodontal health of systemic sclerosis patients compared with non-systemic sclerosis controls. MATERIALS AND METHODS: Online databases were searched for eligible studies on February 24, 2023. The primary outcomes of interest in systemic sclerosis patients and controls included the clinical attachment level, periodontal probing depth, recession depth, plaque index, bleeding on probing score, gingival index, number of teeth with periodontitis, prevalence of periodontitis and gingivitis, and extent and severity of periodontitis. RESULTS: Fourteen studies met inclusion criteria and were incorporated in the qualitative and quantitative analysis. In comparison with the controls, systemic sclerosis patients had a higher prevalence of periodontitis (OR = 7.63 (1.74-33.50), p = 0.04, I2 = 69%), including more severe forms of periodontitis (OR = 6.68 (3.39-13.15), p = 0.85, I2 = 0%), as well as higher periodontal probing depth ((0.88 (0.45-1.31), p = 0.02, I2 = 99%)), clinical attachment level (1.22 (0.8-1.64), p = 0.003, I2 = 98%), and plaque presence (0.83 (0.13-1.53), p = 0.03, I2 = 96%). There was no statistically significant difference in gingival inflammation (1.14 (0.07-2.21), p = 0.04, I2 = 98%). CONCLUSIONS: The systematic review and the meta-analysis showed that systemic sclerosis patients suffer from worse periodontal health than non-systemic sclerosis individuals.

Melatonin Action in Type 2 Diabetic Parotid Gland and Dental Pulp: In Vitro and Bioinformatic Findings.

Type 2 diabetes mellitus (T2DM) is associated with functional deterioration of the salivary gland and dental pulp, related to oxidative stress. The aim was to integrate experimental and bioinformatic findings to analyze the cellular mechanism of melatonin (MEL) action in the human parotid gland and dental pulp in diabetes. Human parotid gland tissue was obtained from 16 non-diabetic and 16 diabetic participants, as well as human dental pulp from 15 non-diabetic and 15 diabetic participants. In human non-diabetic and diabetic parotid gland cells (hPGCs) as well as in dental pulp cells (hDPCs), cultured in hyper- and normoglycemic conditions, glial cell line-derived neurotrophic factor (GDNF), MEL, inducible nitric oxide synthase (iNOS) protein expression, and superoxide dismutase (SOD) activity were measured by enzyme-linked immunosorbent assay (ELISA) and spectrophotometrically. Bioinformatic analysis was performed using ShinyGO (v.0.75) application. Diabetic participants had increased GDNF and decreased MEL in parotid (p < 0.01) and dental pulp (p < 0.05) tissues, associated with increased iNOS and SOD activity. Normoglycemic hDPCs and non-diabetic hPGCs treated with 0.1 mM MEL had increased GDNF (p < 0.05), while hyperglycemic hDPCs treated with 1 mM MEL showed a decrease in up-regulated GDNF (p < 0.05). Enrichment analyses showed interference with stress and ATF/CREB signaling. MEL induced the stress-protective mechanism in hyperglycemic hDPCs and diabetic hPGCs, suggesting MEL could be beneficial for diabetes-associated disturbances in oral tissues.

Patient discomfort and intensity of intraseptal anesthesia of computer-controlled articaine/epinephrine delivery for scaling and root planing.

OBJECTIVES: The primary aim was to assess the pain during intraseptal anesthesia (ISA) administration, as well as during and after scaling and root planing (SRP). The secondary aims pertained to comparing the pain in different jaw regions and evaluating the factors affecting pain during ISA administration, during and after SRP. MATERIAL AND METHODS: ISA was obtained with three different doses of 4% articaine with 1:100,000 epinephrine (4%Ar + Ep) in 360 patients. The pain levels were measured using the visual analogue scales (VAS) for pain intensity during ISA administration (VASa), during SRP (VASi), and after SRP (VASp). These findings were then correlated with periodontal parameters. Regression analysis was performed for pain during ISA, during and after SRP. RESULTS: Anesthesia administration was painful in 80.8% of cases. VASa negatively correlated with pocket depth (PPD). VASi showed no dose-dependency, except in mandibular premolars. VASi negatively correlated with the clinical attachment level (CAL). VASp positively correlated with PPD and CAL. Positive bleeding on probing reduced the chance of pain during ISA administration. Longer anesthesia duration and wider anesthetic field (orally) increased the prospects of painless SRP. CONCLUSIONS: No dose-dependent differences were found regarding patient discomfort and pain intensity of ISA delivery of 4%Ar + Ep for SRP. CLINICAL RELEVANCE: The pain during ISA administration was mild and well tolerated regardless of the anesthetic dose. A lower intensity of pain during SRP can be expected in patients with greater CAL. Post-treatment pain can be anticipated after SRP in the regions with greater PPD and CAL. CLINICAL TRIALS REGISTRATION NUMBER: NCT04392804 (May 9th, 2020).

Salivary Antioxidant Profile in Patients with Systemic Sclerosis and Periodontitis.

Objective: The aim of the present study was to compare periodontal status and antioxidant profile in unstimulated saliva of systemic sclerosis (SSc) patients with periodontitis and systemically healthy periodontitis patients. Design: Twenty patients with established diagnoses of systemic sclerosis and periodontitis (SSc group) and 20 systemically healthy individuals with periodontitis (P group) were enrolled in the study. Clinical periodontal parameters (clinical attachment level (CAL), gingival recession (GR), periodontal probing depth (PPD), and gingival index (GI)) and concentration of uric acid (UA), superoxide dismutase (SOD), and glutathione peroxidase (GPX) in unstimulated saliva samples were assessed. Results: There were significantly higher mean values of CAL (4.8 ± 0.21 mm versus 3.18 ± 0.17 mm; p ≤ 0.001) and GR (1.66 ± 0.90 mm versus 0.46 ± 0.54 mm; p ≤ 0.001) in the SSc group when compared to the P group. Significantly higher level of GPX (p ≤ 0.001) and SOD (p ≤ 0.001) in unstimulated saliva was detected in the SSc group in comparison with the P group. The specific activity of UA did not significantly differ between the two groups (p = 0.083). Conclusion: The results may indicate higher periodontal destruction and antioxidant perturbations in unstimulated saliva of SSc patients with periodontitis compared to systemically healthy periodontitis patients.

Biofilm Formation on the Surfaces of CAD/CAM Dental Polymers.

Dental polymers are now available as monolithic materials which can be readily used in computer-aided design and computer-aided manufacturing (CAD/CAM) systems. Despite possessing numerous advantages over conventionally produced polymers, the polymers produced by either of these systems fail to exhibit immunity to surface microbial adhesion when introduced into the oral environment, leading to the development of oral diseases. The aim of this study was to analyze the biofilm formation of six microorganisms from the oral cavity and its correlation to the surface characteristics of CAD/CAM dental polymers. A total of ninety specimens were divided into three groups: resin-based composite, polymethyl methacrylate, and polyether ether ketone. The experimental procedure included surface roughness and water contact angle measurements, colony forming unit counting, and scanning electron microscopy analysis of biofilm formed on the surface of the tested materials. The data were analyzed using the Kruskal-Wallis test, with a Dunn's post hoc analysis, and one way analysis of variance, with a Tukey's post hoc test; the correlation between the measurements was tested using Spearman's correlation coefficient, and descriptive statistics were used to present the data. Despite using the same manufacturing procedure, as well as the identical manufacturer's finishing and polishing protocols, CAD/CAM dental polymers revealed significant differences in surface roughness and water contact angle, and the increased values of both parameters led to an increase in biofilm formation on the surface of the materials. The CAD/CAM resin-based composite showed the lowest number of adhered microorganisms compared to CAD/CAM polymethyl methacrylate and CAD/CAM polyether ether ketone.

Efficacy of intraseptal anesthesia obtained by computer-controlled articaine with epinephrine delivery in scaling and root planing.

OBJECTIVES: The primary aim of this study was to compare the anesthetic efficacy of the intraseptal anesthesia (ISA) obtained with three doses of 4% articaine with 1:100,000 epinephrine (4%Ar + Ep) for scaling and root planing (SRP), using a computer-controlled local anesthetic delivery system (CCLADS). The secondary aims were to compare the clinical anesthetic parameters in relation to different jaw regions and examine the possible influence of sex and smoking habits on them. MATERIALS AND METHODS: SRP under ISA obtained with different doses (0.1 ml, 0.2 ml, and 0.3 ml) of 4%Ar + Ep was performed in 360 patients. The success rate, onset, duration of soft tissue anesthesia, and the anesthetic field widths were recorded by pinprick testing. RESULTS: The anesthesia success was high (90-95%). The onset was immediate. The duration and anesthetic field widths showed a dose-related significance, however, without a consistent sex-related or smoking-related significance. The multiple logistic regression analysis revealed a twofold higher chance of anesthesia success by increasing the dose and increased bleeding on probing-related and female sex-reduced probability of anesthesia success. CONCLUSIONS: ISA obtained with 0.3 ml of 4%Ar + Ep delivered by a computer-controlled local anesthetic delivery system provided a high anesthetic success and the adequate clinical anesthetic parameters for SRP in all regions of both jaws. CLINICAL RELEVANCE: ISA obtained with 4%Ar + Ep provides an effective anesthesia for SRP. The anesthetic success rate may be reduced in the presence of gingival inflammation and in females as well. The study was registered in a Clinical Trials database (NCT04392804, registration date May 9, 2020).

Effect of section thickness on cone beam computed tomography-based measurements of intrabony defects compared with clinical measurements.

BACKGROUND: It is unknown whether cone beam computed tomography (CBCT) image reconstruction characteristics, including section thickness, may affect linear bone measurements of periodontal intrabony defects. The aim of this study was to compare intrasurgical and CBCT-based linear measurements of intrabony defects focusing on CBCT section thickness. METHODS: Sixty-six intrabony defects were assessed in 21 patients with chronic generalized severe periodontitis. Linear measurements of alveolar bone (radiographic bone level [rBL]), assessed in CBCT images at diverse section thicknesses: 0.25 mm (voxel size), 1 mm, and 3 mm, were compared with clinical bone level (cBL) measurements obtained intrasurgically. To provide identical reference points for rBL and cBL measurements, individually adjusted grooves on the reference stent were prepared for each periodontal defect site. CBCT measurements were performed in two rounds by two trained observers. Observer agreement was assessed by intraclass correlation coefficients (ICC). ANOVA assessed the difference among cBL and rBL at different section thicknesses. RESULTS: Intra- and inter-observer agreement was excellent (ICC >0.99) and highly significant independent of the observer, evaluation round, and CBCT section thickness. Mean rBL in the diverse CBCT section thicknesses was very close to that measured clinically (cBL). There was no statistically significant difference between cBL and rBL for any section thickness, neither for the overall evaluated sites, nor the maxilla or mandible separately. CONCLUSIONS: No statistically significant difference between clinical and radiographic bone level for 0.25-, 1-, and 3-mm CBCT section thicknesses were observed when assessing intrabony periodontal defects.

The use of collagen porcine dermal matrix and connective tissue graft with modified coronally advanced tunnel technique in the treatment of multiple adjacent type I gingival recessions: A randomized, controlled clinical trial.

OBJECTIVE: To assess the clinical efficacy and esthetic outcome of porcine-derived dermal collagen matrix in comparison with connective tissue graft in the treatment of multiple adjacent gingival recessions (MAGR), 6 and 12 months after the surgery. MATERIALS AND METHODS: Twenty patients with bilateral type I MAGR were treated randomly with porcine-derived dermal collagen matrix (test site) or connective tissue graft (control site) in combination with a modified coronally advanced tunnel technique. The primary objectives were to evaluate the mean and complete root coverage. The secondary objectives were to assess keratinized tissue width, gingival thickness gain, and root coverage esthetic score. RESULTS: Six and 12 months postoperatively, both groups achieved significant improvements in all clinical parameters compared to baseline, with no statistically significant differences between the groups. Mean root coverage change (Δ12m - 6 m) was statistically significant between the groups in favor of connective tissue graft, and twice as many patients exhibited a complete coverage of all recessions in the control group than the test group. CONCLUSION: The porcine-derived dermal collagen matrix combined with a modified coronally advanced tunnel technique resulted in satisfactory clinical and esthetic outcomes, which were similar to connective tissue graft. CLINICAL SIGNIFICANCE: Porcine-derived dermal collagen matrix (XDM) may be proposed as a substitute for connective tissue graft in multiple adjacent recession treatment due to successful root coverage, a significant increase of gingival thickness, and high esthetic outcomes. The clinical benefits for the use of XDM could be: (a) second surgical wound avoidance, (b) patient discomfort decrease, and (c) lower complications' rate.

MicroRNA-146a and microRNA-155 as novel crevicular fluid biomarkers for periodontitis in non-diabetic and type 2 diabetic patients.

AIM: Recent studies point at the crucial role of epigenetic mechanisms in the development of multifactorial diseases such as periodontitis and diabetes mellitus (DM) type 2. In addition, circulatory microRNAs (miRs) have emerged as novel biomarkers for various diseases. Aim of this study was to investigate the levels of miR-146a and miR-155 and superoxide dismutase (SOD) activity in gingival crevicular fluid (GCF) of periodontitis patients with (CPDM) and without (CP) DM type 2 as well as in periodontally healthy, control groups (PHDM and PH, respectively). MATERIAL AND METHODS: miR modulation was analysed using quantitative real-time PCR while SOD activity was measured spectrophotometrically. RESULTS: The upregulation of miR-146a and miR-155 was observed in CP and CPDM patients' baseline, while the levels decreased after 6 weeks of the non-surgical therapy to the levels comparable to PH and PHDM, respectively. Expression levels of miRs positively correlated with SOD activity. Levels of miR-146a were higher in PHDM compared to PH patients. Multivariate analysis revealed that levels of miR-146a and miR-155 were significantly associated with periodontitis when adjusting for age and gender. CONCLUSIONS: miR-146a and miR-155 may be considered as possible novel biomarkers for periodontitis in non-diabetic and type 2 diabetic patients.

Cone-Beam Computed Tomography for Detection of Intrabony and Furcation Defects: A Systematic Review Based on a Hierarchical Model for Diagnostic Efficacy.

BACKGROUND: The aim of this review is to assess the diagnostic efficacy of cone-beam computed tomography (CBCT) for the diagnosis of and/or treatment planning for intrabony and furcation defects, using a well-known six-tiered hierarchical model for diagnostic efficacy. METHODS: The MEDLINE, EMBASE, and Cochrane Library bibliographic databases were searched until August 2015 for studies evaluating CBCT imaging for the diagnosis of and/or treatment planning for intrabony and/or furcation defects. The search strategy was restricted to English language publications using the combination of MeSH terms, free terms, and key words. RESULTS: The search strategy yielded 16 publications that qualitatively or quantitatively evaluated the use of CBCT for the detection of intrabony and/or furcation defects and how CBCT influenced the diagnosis and/or treatment plan. According to Quality Assessment of Studies of Diagnostic Accuracy-2, all included studies were medium to low risk of bias. The review identified only one study that investigated the societal efficacy, and none evaluated the patient outcome efficacy or therapeutic efficacy. One study investigated the diagnostic thinking efficacy. All other included studies investigated the diagnostic accuracy of CBCT. CONCLUSIONS: From the assessed studies, it can be concluded that there is not sufficient scientific evidence to justify the use of CBCT for the diagnosis of and/or treatment planning for intrabony and furcation defects. Furthermore, the effectiveness of CBCT for such diagnostic tasks has been assessed only at low diagnostic efficacy levels.

Multiple Major and Minor Anomalies Associated With Klippel-Feil Syndrome: A Case Report.

Klippel-Feil syndrome is defined as congenital fusion of two or more cervical vertebrae. In this article, we report a 55-year-old male patient with one-year history of neck pain, headaches, and one episode of syncope after a severe trauma. X-rays and magnetic resonance imaging of cervical spine revealed fused vertebral bodies of C2-C5. The major anomalies associated with Klippel-Feil syndrome (small stature, thoracic kyphoscoliosis, lumbar scoliosis, restricted opening mouth, and bilateral sensorineural hearing loss) as well as multiple minor anomalies (mild face asymmetry, high arched palate, rhinoscoliosis, high nasal bridge, inclined septi nasi, and thin upper lip) were detected. This is a rare case describing the anomalies of the nose in Klippel-Feil syndrome patients. Our patient had no central cord impairment following a severe trauma.

Clinical and CBCT-based diagnosis of furcation involvement in patients with severe periodontitis.

OBJECTIVE: The aim of this study was to compare the use of periodontal probing and cone beam computed tomography (CBCT) images in the diagnosis of furcation involvement (FI) in patients with chronic generalized severe periodontitis. METHOD AND MATERIALS: Fifteen patients with chronic generalized severe periodontitis were included in this study. In total, 174 furcation sites (all in molar teeth) were analyzed. FI was assessed at three sites (buccal, mesiopalatal, and distopalatal) of maxillary molars, and at two sites (buccal and oral) of mandibular molars. FI was assessed both clinically (periodontal probing) and on CBCT images, using a dichotomous scale (present/absent). The agreement between clinical and CBCT-based findings was calculated. RESULTS: FI were more often detected by means of CBCT than by means of clinical examination. Agreement between the evaluation methods was present in 46.9% of cases (63.3% in maxilla, 45.0% in mandible). FI detected clinically was confirmed by means of CBCT in 24% of the evaluated sites. The largest agreement (73.7%) in FI detection was found in the distopalatal maxillary sites between CBCT and clinical probing. The smallest agreement (36.6%) was found in the buccal sites of the mandibular molars, in which 63.3% of FI were detected using CBCT only, but not clinically. CONCLUSION: The number of FI detected by means of CBCT was larger than by means of periodontal probing. In those cases in which chronic generalized severe periodontitis is clinically diagnosed, and surgical treatment is necessary, CBCT may be suggested as an adjunct tool for FI assessment.

Receptor activator of nuclear factor kappa B (RANK) as a determinant of peri-implantitis.

BACKGROUND/AIM: Peri-implantitis presents inflammatory process that affects soft and hard supporting tissues of osseointegrated implant based on inflammatory osteoclastogenesis. The aim of this study was to investigate whether receptor activator of nuclear factor kappa B (RANK) concentrations in peri-implant crevicular fluid could be associated with clinical parameters that reflect inflammatory nature of peri-implantitis. METHODS: The study included 67 patients, 22 with diagnosed peri-implantitis, 22 persons with healthy peri-implant tissues and 23 patients with periodontitis. Clinical parameters from each patient were recorded and samples of peri-implant/gingival crevicular fluid were collected for the enzyme-linked immunosorbent assay (ELISA) analysis. RESULTS: RANK concentration was significantly increased in samples from the patients with peri-implantitis when compared to healthy implants (p < 0.0001), where the average levels were 9 times higher. At the same time RANK concentration was significantly higher in peri-implantitis than in periodontitis sites (p < 0.0001). In implant patients pocket depths and bleeding on probing values were positively associated with high RANK concentrations (p < 0.0001). CONCLUSION: These results revealed association of increased RANK concentration in samples of peri-implant/gingival crevicular fluid with peri-implant inflammation and suggests that RANK could be a pathologic determinant of peri-implantitis, thereby a potential parameter in assessment of peri-implant tissue inflammation and a potential target in designing treatment strategies.

Bone loss biomarkers associated with peri-implantitis. A cross-sectional study.

AIM: To investigate the levels of biomarkers associated with osteoclastogenesis in patients suffering peri-implantitis and to compare them with levels in healthy peri-implant sites and severe chronic periodontitis. MATERIAL AND METHODS: Peri-implant/gingival crevicular fluid samples and clinical parameters including: bleeding on probing, modified Plaque Index (PlI), pocket depth and clinical attachment level were collected from 70 patients (23 with peri-implantitis, 25 with healthy peri-implant tissues and 22 with severe chronic periodontitis). The concentrations of sRANKL, RANK and OPG were evaluated using enzyme-linked immunosorbent assays; they were compared between the groups and correlated with the clinical findings. RESULTS: sRANKL (P = 0.01), RANK (P = 0.01) and OPG (P = 0.03) concentrations were significantly higher in peri-implantitis sites when compared to those in healthy implant sites, although differences in the sRANKL/OPG ratio were not statistically significant. In these sites all three markers were significantly correlated with the clinical parameters, with exception of OPG/PI correlation that remained insignificant (P = 0.121). When comparing peri-implantitis and periodontitis findings, RANK was significantly higher in peri-implantitis sites whereas, sRANKL (P = 0.03) and sRANKL/OPG ratio (P = 0.004) were significantly higher in periodontitis sites. Among periodontitis and healthy implant sites the same differences have been observed for both sRANKL (P = 0.000) and sRANKL/OPG ratio (P = 0.000), furthermore RANK was higher in periodontitis sites as well (P = 0.010). CONCLUSION: The findings of this preliminary study on a relatively small sample size suggest that the PICF levels of biomarkers sRANKL, RANK, and OPG are associated with peri-implant tissue destruction and the pattern of these biomarkers differed when compared to periodontitis.

Association of fibronectin with hypogalactosylated immunoglobulin G in gingival crevicular fluid in periodontitis.

BACKGROUND: Fibronectin (FN) can bind to immunoglobulins (Ig), influencing both the normal clearance and abnormal deposition of circulating immune complexes. This study focuses on the possible interaction between FN and IgG present in gingival crevicular fluid (GCF) of periodontitis patients and periodontally healthy controls, with the aim to determine whether such interaction may be connected with the glycosylation profile of IgG and, thus, consequentional in accumulation of proinflammatory IgG in periodontal pockets. METHODS: The study included 30 patients with initial or advanced periodontitis, and 13 periodontally healthy subjects. GCF IgG was purified and analyzed for the presence of FN and its fragments and for galactose expression. RESULTS: IgG isolated from GCF contained FN, which was bound to the IgG heavy chains. IgG from GCF of advanced periodontitis patients contained high levels of hypogalactosylated IgG, and bound more FN than IgG from GCF of the other two groups, which contained low levels of this glycoform. FN was in a degraded form in GCF from all studied patients, and a fragment of 48- to 53-kDa molecular mass seemed to be the sole one linked to IgG. CONCLUSIONS: IgG and the FN fragment of 48 to 53 kDa in GCF of periodontitis patients and periodontally healthy subjects are physically connected. This fragment was linked to the heavy chains of IgG and the reaction seemed to be particularly intensive with IgG from advanced periodontitis, which contained significantly less galactose in its sugar chains.

[Identification of periodontopathogen microorganisms by PCR technique].

INTRODUCTION: Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE: The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD: Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS: In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION: The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria.