A rapid, non-invasive tool for periodontitis screening in a medical care setting.

BACKGROUND: Since periodontitis is bi-directionally associated with several systemic diseases, such as diabetes mellitus and cardiovascular diseases, it is important for medical professionals in a non-dental setting to be able examine their patients for symptoms of periodontitis, and urge them to visit a dentist if necessary. However, they often lack the time, knowledge and resources to do so. We aim to develop and assess "quick and easy" screening tools for periodontitis, based on self-reported oral health (SROH), demographics and/or salivary biomarkers, intended for use by medical professionals in a non-dental setting. METHODS: Consecutive, new patients from our outpatient clinic were recruited. A SROH questionnaire (8 questions) was conducted, followed by a 30 s oral rinse sampling protocol. A complete clinical periodontal examination provided the golden standard periodontitis classification: no/mild, moderate or severe periodontitis. Total periodontitis was defined as having either moderate or severe. Albumin and matrix metalloproteinase-8 concentrations, and chitinase and protease activities were measured in the oral rinses. Binary logistic regression analyses with backward elimination were used to create prediction models for both total and severe periodontitis. Model 1 included SROH, demographics and biomarkers. The biomarkers were omitted in the analysis for model 2, while model 3 only included the SROH questionnaire. The area under the receiver operating characteristic curves (AUROCC) provided the accuracy of each model. The regression equations were used to create scoring algorithms, composed of the remaining predictors, each with its own weight. RESULTS: Of the 156 patients participating in this study, 67% were classified with total periodontitis and 33% had severe periodontitis. The models for total periodontitis achieved an AUROCC of 0.91 for model 1, 0.88 for model 2 and 0.81 for model 3. For severe periodontitis, this was 0.89 for model 1, 0.82 for model 2 and 0.78 for model 3. The algorithm for total periodontitis (model 2), which we consider valid for the Dutch population, was applied to create a freely accessible, web-based screening tool. CONCLUSIONS: The prediction models for total and severe periodontitis proved to be feasible and accurate, resulting in easily applicable screening tools, intended for a non-dental setting.

On site visual detection of Porphyromonas gingivalis related periodontitis by using a magnetic-nanobead based assay for gingipains protease biomarkers.

Porphyromonas gingivalis (P. gingivalis) is a pathogen causing periodontitis. A rapid assay is described for the diagnosis of periodontal infections related to P. gingivalis. The method is making use of gingipains, a group of P. gingivalis specific proteases as a detection biomarker. Magnetic-nanobeads were labeled with gingipain-specific peptide substrates and immobilized on a gold biosensing platform via gold-thiol linkage. As a result of this, the color of the gold layer turns black. Upon cleavage of the immobilized substrates by gingipains, the magnetic-nanobeads-peptide fragments were attracted by a magnet so that the golden surface color becomes visible again. This assay is highly sensitive and specific. It is capable of detecting as little as 49 CFU·mL-1 of P. gingivalis within 30 s. Examination of periodontitis patients and healthy control saliva samples showed the potential of the assay. The simplicity and rapidity of the assay makes it an effective point-of-care device. Graphical abstract Schematic of the assay for the detection of P. gingivalis proteases as one of the promising biomarkers associated with periodontal diseases.

Reduced platelet hyper-reactivity and platelet-leukocyte aggregation after periodontal therapy.

BACKGROUND: Platelets from untreated periodontitis patients are hyper-reactive and form more platelet-leukocyte complexes compared to cells from individuals without periodontitis. It is not known whether the improvement of the periodontal condition achievable by therapy has beneficial effects on the platelet function. We aimed to assess the effects of periodontal therapy on platelet reactivity. METHODS: Patients with periodontitis (n = 25) but unaffected by any other medical condition or medication were included and donated blood before and after periodontal therapy. Reactivity to ADP or oral bacteria was assessed by flow cytometric analysis of membrane markers (binding of PAC-1, P-selectin, CD63) and platelet-leukocyte complex formation. Reactivity values were expressed as ratio between the stimulated and unstimulated sample. Plasma levels of soluble (s) P-selectin were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Binding of PAC-1, the expression of P-selectin and CD63 in response to the oral bacterium P. gingivalis were lower at recall (1.4 ± 1.1, 1.5 ± 1.2, and 1.0 ± 0.1) than at baseline (2.7 ± 4.1, P = 0.026, 6.0 ± 12.5, P = 0.045, and 2.7 ± 6.7, P = 0.042, respectively). Formation of platelet-leukocyte complexes in response to P. gingivalis was also reduced at recall compared to baseline (1.2 ± 0.7 vs. 11.4 ± 50.5, P = 0.045). sP-selectin levels were significantly increased post-therapy. CONCLUSIONS: In periodontitis patients, the improvement of the periodontal condition is paralleled by a reduction in platelet hyper-reactivity. We suggest that periodontal therapy, as an intervention for improved oral health, can facilitate the management of thrombotic risk, and on the long term can contribute to the prevention of cardiovascular events in patients at risk. TRIAL REGISTRATION: Current Controlled Trials identifier ISRCTN36043780. Retrospectively registered 25 September 2013.

Effect of periodontal therapy with systemic antimicrobials on parameters of metabolic syndrome: A randomized clinical trial.

AIM: To investigate the effect of basic periodontal therapy (BPT) with antimicrobials (AM) on the parameters of metabolic syndrome (MetS) (waist circumference, systolic/diastolic blood pressure [BP], HDL-cholesterol, triglycerides, glucose). METHODS: One hundred and ten periodontitis patients without known comorbidities and unaware of possible MetS were randomly assigned to BPT (n = 56) or BPT+AM (n = 54) and followed for 12 months post-therapy. Number of patients with undiagnosed MetS was also determined. RESULTS: In all patients, the periodontal condition improved; however, the BTP+AM group showed greater pocket depth reduction than the BPT group. Post-therapy, systolic BP (p < .05) and triglycerides (p < .05) reduced significantly during the follow-up period. No significant differences could be assessed between the BPT+AM and BPT group. Despite the absence of self-reported comorbidities, 27.2% (n = 30) periodontitis patients fulfilled the criteria of MetS at baseline. After therapy, this proportion changed to 14.5% at 3 months (p = .007), to 17.3% at 6 months (p = .017) and to 21.8% at 12-month follow-up (p = .383). CONCLUSION: Although a reduction in systolic BP and triglycerides and a temporarily improvement in the whole metabolic status were observed, the use of antimicrobials in conjunction with BTP does not yield any additional improvement in the parameters of MetS.

Local disinfection with sodium hypochlorite as adjunct to basic periodontal therapy: a randomized controlled trial.

AIM: To investigate the clinical and microbiological effects of local disinfection with 0.5% sodium hypochlorite (NaOCl) with or without systemic antimicrobials (amoxicillin and metronidazole, AM) during basic periodontal therapy (BPT). MATERIAL AND METHODS: In a randomized clinical trial (four groups), 110 chronic periodontitis patients received BPT plus local irrigation with saline (BPT + S), local disinfection with NaOCl (BPT + DIS), BPT + DIS + AM or BPT + S + AM. The outcome was analysed at baseline, 3, 6 and 12 months. RESULTS: There was no difference in clinical attachment level gain at 12 months between the four groups. BPT + DIS showed no additional improvement compared to BPT + S; BPT + DIS + AM showed fewer sites with probing pocket depth (PPD) ≥7 mm versus BPT + S only up to 6 months (p = 0.037). In factorial analyses, additional clinical reduction for PPD (p = 0.023) and number of sites with PPD ≥5 (p = 0.007), ≥6 (p = 0.002) and ≥7 mm (p < 0.001) were found when AM was added to BPT, but not when DIS was applied. In all groups, a comparable decrease in targeted bacteria was found. AM caused adverse events in 22% of the patients. CONCLUSION: Local disinfection with NaOCl, also in combination with AM, showed, after 1-year follow-up, no additional clinical and microbiological effects compared to BPT alone.

Colorimetric Assay for the Detection of Typical Biomarkers for Periodontitis Using a Magnetic Nanoparticle Biosensor.

Periodontitis is a chronic disease which affects at least 10% of the population. If untreated, periodontitis can lead to teeth loss. Unfortunately, current diagnostic tests are limited in their sensitivity and specificity. In this study, a novel multiplex hand-held colorimetric diagnostic biosensor, using two typical inflammatory salivary biomarkers, Human Neutrophil Elastase (HNE) and Cathepsin-G, was constructed as proof of concept to potentially detect periodontitis. The biosensing method was based on the measurement of proteolytic activity using specific proteases probes. These probes consist of specific proteases substrates covalently bound to a magnetic bead from one end and to the gold sensor surface by the other end. When intact, this renders the golden sensor black. Upon proteolysis, the cleaved magnetic beads will be attracted by an external magnet revealing the golden color of the sensor surface observable by the naked eye. The biosensor was capable of specific and quantitative detection of HNE and Cathepsin-G in solution and in spiked saliva samples with a lower detection limit of 1 pg/mL and 100 fg/mL for HNE and Cathepsin-G, respectively. Examination of periodontitis patients' sample and a healthy control showed the potential of the multiplex biosensor to detect the presence of HNE and Cathepsin-G activity in situ. This approach is anticipated to be a useful biochip array amenable to low-cost point-of-care devices.

Subgingival microbiome in smokers and non-smokers in periodontitis: an exploratory study using traditional targeted techniques and a next-generation sequencing.

AIM: To compare the results of two targeted techniques to an open-ended technique in periodontitis patients, differentiated on the basis of smoking habit. MATERIALS & METHODS: Thirty periodontitis patients (15 smokers and 15 non-smokers) provided subgingival plaque samples for 16S rRNA gene amplicon sequencing, culturing and quantitative polymerase chain reaction (qPCR). RESULTS: No differences were found in the composition of the subgingival microbiome between smokers and non-smokers with culture and qPCR. With pyrosequencing, operational taxonomic units (OTUs) classified to genera Fusobacterium, Prevotella and Selenomonas were more abundant in smokers, while OTUs belonging to the genera Peptococcus and Capnocytophaga were more abundant in non-smokers. Principal coordinate analysis identified two clusters; one was composed mainly of smokers (80%) and revealed significantly lower taxonomic diversity, higher attachment loss and higher proportion of the genera Fusobacterium, Paludibacter and Desulfobubus. CONCLUSION: In periodontitis, there is a difference in the composition of the subgingival microbiome between smokers and non-smokers, as revealed by pyrosequencing. This difference was not identified by the targeted techniques. Low taxonomic diversity was associated with higher disease severity, especially in smokers. This supports the hypothesis of the ecological microbial-host interaction in the severity of periodontal disease.

Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis.

Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluorogenic dipeptides that contain d-amino acids, led to the discovery of five P. gingivalis-specific substrates. Due to the presence of lysine and arginine residues in these substrates, it was hypothesized that the cleavage was mediated by the gingipains, a group of P. gingivalis-specific proteases. This hypothesis was confirmed by the observation that P. gingivalis gingipain knockout strains demonstrated clearly impaired substrate cleavage efficacy. Further, proteolytic activity on the substrates was increased by the addition of the gingipain stimulators dithiothreitol and l-cysteine and decreased by the inhibitors leupeptin and N-ethylmaleimide. Screening of saliva and gingival crevicular fluid of periodontitis patients and healthy controls showed the potential of the substrates to diagnose the presence of P. gingivalis proteases. By using paper points, a sensitivity of approximately 10(5) CFU/ml was achieved. P. gingivalis-reactive substrates fully composed of l-amino acids and Bz-l-Arg-NHPhNO(2) showed a relatively low specificity (44 to 85%). However, the five P. gingivalis-specific substrates that each contained a single d-amino acid showed high specificity (96 to 100%). This observation underlines the importance of the presence of d-amino acids in substrates used for the detection of bacterial proteases. We envisage that these substrates may improve the specificity of the current enzyme-based diagnosis of periodontitis associated with P. gingivalis.

Vitamin C in plasma and leucocytes in relation to periodontitis.

AIM: To test the hypothesis that vitamin C concentrations in plasma, polymorphonuclear neutrophilic leucocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) are lower in periodontitis patients compared with healthy controls. METHODS: Twenty-one untreated periodontal patients and 21 healthy controls matched for age, gender, race and smoking habits were selected. Dietary vitamin C intake was assessed by a self-administered dietary record. Fasting blood samples were obtained and analysed for vitamin C concentrations in plasma, PMNs and PBMCs by means of high-pressure liquid chromatography (HPLC). RESULTS: Plasma vitamin C was lower in periodontitis patients compared with controls (8.3 and 11.3 mg/l, respectively, p = 0.03). Only in the control group a positive correlation was present between vitamin C intake and plasma values. No differences could be assessed between patients and controls regarding vitamin C dietary intake and levels in PMNs and PBMCs. In the patient group, pocket depth appeared to be negatively associated with the vitamin C concentration in PMNs. CONCLUSION: Although the relationship between low plasma vitamin C levels and periodontitis is clear, the disease cannot be explained by insufficient vitamin C storage capacity of leucocytes; the question remains through which mechanism low plasma vitamin C levels are related to periodontitis.

Overview of the Effect of Different Regenerative Materials in Class II Furcation Defects in Periodontal Patients.

The aim of this review was to give an overview of the outcomes of the use of different regenerative materials to treat molars with class II furcation defects in patients with periodontitis in comparison with open flap debridement (OFD). A search of five databases (PubMed-Medline, Embase, Cochrane, Scopus and Web of Science) was conducted up to and including January 2022. According to the PICOS guidelines, only randomized control trials (S) considering periodontal patients with at least one molar with a class II furcation involvement (P) treated with regenerative materials (I) in comparison to OFD as control treatment (C) and a minimum follow-up period of 6 months were included. Vertical clinical attachment level (VCAL) was considered as the primary outcome (O), while horizontal clinical attachment level (HCAL), horizontal probing depth (HPD) and vertical probing depth (VPD) were considered as secondary outcomes. The search through the databases initially identified 1315 articles. Only 25 of them met the eligibility criteria and were included. The studies were grouped in four macro-categories according to the material used: absorbable and non-absorbable membranes, blood derivatives and a combination of different materials. The greater part of the included studies reported a statistically significant difference in using regenerative materials when compared to OFD. The blood derivatives groups reported a range of mean changes in VCAL of 0.86-4.6 mm, absorbable membrane groups reported -0.6-3.75 mm, non-absorbable membranes groups reported -2.47-4.1 mm, multiple materials groups reported -1.5-4.87 mm and enamel matrix derivatives reported a mean change in VCAL of 1.45 mm. OFD showed a range of mean VCAL changes of -1.86-2.81 mm. Although no statistical analysis was performed, the use of regenerative materials may be considered moderately beneficial in the treatment of molars with grade II furcation involvement. However, the substantial heterogeneity in the protocols' design does not allow us to draw definitive conclusions. In addition, low levels of evidence for morbidity and patient-centered outcomes were reported.

Association of serum immunoglobulin G (IgG) levels against two periodontal pathogens and prothrombotic state: a clinical pilot study.

OBJECTIVE: Periodontitis is associated with cardiovascular diseases (CVD). In our previous studies a prothrombotic state has been observed in periodontitis, which contributes to the risk of CVD. The aim of this study was to investigate whether serum IgG levels against Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) in periodontitis were associated with a prothrombotic state. MATERIALS AND METHODS: Patients with moderate (n = 38) and severe periodontitis (n = 30) and controls (n = 24) were recruited. We explored correlations between serum anti-Aa and anti-Pg IgG and plasma levels of markers of prothrombotic state (von Willebrand Factor [vWF], prothrombin fragment 1+2 [F1+2], plasminogen activator inhibitor-1 [PAI-1] and D-dimer). Multivariate analyses were performed considering several major potential contributing factors. RESULTS: Periodontitis patients showed higher anti-Aa IgG (p = 0.015) than controls but not for Pg (p = 0.320). In periodontitis patients, body mass index and anti-Aa IgG showed a positive correlation with vWF (ß = 0.297, p = 0.010 and ß = 0.248, p = 0.033 respectively). CONCLUSIONS: In periodontitis, infection with Aa together with other well accepted risk factors for CVD, may play a role in increasing the risk for prothrombotic state.

Use of Platelet-Rich Fibrin in the Treatment of Grade 2 Furcation Defects: Systematic Review and Meta-Analysis.

In periodontitis patients, furcation defects are crucial sites to regenerate due to their complex anatomy. Various modern surgical techniques and use of biomaterials have been suggested in the literature. Among all, platelet-rich fibrin (PRF) has potential in tissue regeneration thanks to its role in the release of growth factors. Therefore, the purpose of this study was to evaluate the beneficial effect of the addition of PRF to open flap debridement (OFD) or as an adjuvant to other biomaterials such as bone grafts in the treatment of grade 2 mandibular furcation defects. Systematic research was carried out on the databases Medline, Scopus, Embase, and Cochrane Library and registered on PROSPERO (CRD42020167662). According to the PICO guidelines by Cochrane, randomized trials and prospective non-randomized trials were evaluated, with a minimum follow-up period of 6 months. The inclusion criteria were the absence of systemic diseases, non-smoking patients, and a population aged from 18 to 65 years. Vertical pocket probing depth (PPD), vertical clinical attachment level (VCAL), and gingival recession (REC) were the primary outcomes. Vertical furcation depth (VFD), and the percentage of bone defect fill (%v-BDF) were considered as secondary outcomes. A meta-analysis of the primary and secondary outcomes was performed. Publication bias was assessed through a funnel plot. Eighty-four articles were initially extracted. Eight randomized clinical trials were analyzed according to the exclusion and inclusion criteria. The Quality assessment instrument (QAI) revealed four articles at low risk of bias, one at moderate, and three at high risk of bias. The metanalysis showed significant data regarding PPD, VCAL, VFD and %v-BDF in the comparison between PRF + OFD vs. OFD alone. The adjunct of PRF to a bone graft showed a significant difference for VCAL and a not statistically significant result for the other involved parameters. In conclusion, the adjunctive use of PRF to OFD seems to enhance the periodontal regeneration in the treatment of grade 2 furcation defects. The combination of PRF and bone graft did not show better clinical results, except for VCAL, although the amount of literature with low risk of bias is scarce. Further well-designed studies to evaluate the combination of these two materials are therefore needed.

Periodontal therapy increases neutrophil extracellular trap degradation.

In periodontitis, polymorphonuclear leucocytes (PMNs) are activated. They entrap and eliminate pathogens by releasing neutrophil extracellular traps (NETs). Abnormal NET degradation is part of a pro-inflammatory status, affecting co-morbidities such as cardiovascular disease. We aimed to investigate the ex vivo NET degradation capacity of plasma from periodontitis patients compared to controls (part 1) and to quantify NET degradation before and after periodontal therapy (part 2). Fresh NETs were obtained by stimulating blood-derived PMNs with phorbol 12-myristate 13-acetate. Plasma samples from untreated periodontitis patients and controls were incubated for 3 h onto freshly generated NETs (part 1). Similarly, for part 2, NET degradation was studied for 91 patients before and 3, 6 and 12 mo after non-surgical periodontal therapy with and without adjunctive systemic antibiotics. Finally, NET degradation was fluorospectrometrically quantified. NET degradation levels did not differ between periodontitis patients and controls, irrespective of subject-related background characteristics. NET degradation significantly increased from 65.6 ± 1.7% before periodontal treatment to 75.7 ± 1.2% at 3 mo post periodontal therapy, and this improvement was maintained at 6 and 12 mo, irrespective of systemic usage of antibiotics. Improved NET degradation after periodontitis treatment is another systemic biomarker reflecting a decreased pro-inflammatory status, which also contributes to an improved cardiovascular condition.

The link between periodontitis and erectile dysfunction: a review.

Introduction Consistent evidence has shown that periodontitis can be considered a risk factor for the development of vascular complications such as myocardial infarction and cerebrovascular accident. The majority of cases of erectile dysfunction (ED) are considered to result from the complication of vascular impairments. Therefore, it is conceivable to hypothesise that periodontitis can also be associated with ED.Aims To determine whether a possible link between periodontitis and ED exists by reviewing and presenting the current available evidence.Methods Current, up to June 2018, case-control studies, randomised controlled trials (RCT) and meta-analyses were reviewed.Results Nine case-control studies and three meta-analyses found significant positive associations between these two conditions, with odds ratios ranging from 1.53 to 5.94. Furthermore, one RCT found a significant improvement in subjective measurements of ED in patients treated for periodontitis.Conclusions The current associations must be interpreted with caution because of the considerable heterogeneity of the cross-sectional investigations and the short-term character of the only RCT included. Nevertheless, the preliminary results can be taken into consideration for the general physician or the specialist in the motivation of the male patient to visit dental professionals and, if indicated, treated for periodontitis, which may help in managing the vasculogenic form of impotence.

Comment on "Aggregatibacter actinomycetemcomitans-induced hypercitrullination links periodontal infection to autoimmunity in rheumatoid arthritis".

The link between rheumatoid arthritis and exposure to a bacterial toxin was not found in a population of rheumatoid arthritis patients from Netherlands.

A pilot study into measurements of markers of atherosclerosis in periodontitis.

BACKGROUND: Periodontitis may be a possible risk factor for atherosclerosis. The current pilot study explored arterial wall thickness and other variables associated with atherosclerosis in healthy subjects with and without periodontitis. METHODS: Patients with moderate (N = 34) and severe periodontitis (N = 15) and controls (N = 14) were recruited. Intima media thickness (IMT) of the common carotid arteries (CCA), internal carotid arteries (ICA), and bifurcations of carotid arteries (BCA) was estimated bilaterally using B-mode ultrasound. An overall IMT was calculated as the mean of these six measurements. C reactive protein (CRP), fibrinogen, and von Willebrand factor (vWf) were measured in plasma as indicators of systemic inflammation and atherosclerotic disease. Microalbuminuria was determined as a marker of endothelial cell dysfunction. RESULTS: IMT for CCA were 0.64, 0.68, and 0.69 mm for control, moderate, and severe periodontitis, respectively (not significant). IMT for BCA did not vary among groups. IMT of ICA was largest for severe periodontitis (0.81 mm); corresponding values for controls and moderate periodontitis were 0.58 and 0.55 mm, respectively (P= 0.023). Severe periodontitis patients had an overall IMT of 0.76 mm, while moderate periodontitis patients and controls had lower values (0.64 and 0.65 mm, respectively; P= 0.153). After adjusting for potential confounding factors, the increased IMT for ICA in severe periodontitis was also significant (Padj = 0.040). CRP (P= 0.020, Padj = 0.050) and vWf (P= 0.019, Padj = 0.013) were higher in periodontitis than controls; microalbuminuria was not different between groups. Power calculations suggest that a 4-fold expansion of the severe patient and control groups will result in a high chance (power level 80%) that a clinically significant association between the overall IMT and periodontitis will be observed. CONCLUSION: The present pilot study indicates that a full study investigating the relationship between periodontitis and atherosclerosis is warranted.

A Lead ANRIL Polymorphism Is Associated with Elevated CRP Levels in Periodontitis: A Pilot Case-Control Study.

Elevated high sensitive C-reactive protein (hsCRP) is a marker for systemic inflammation and a risk marker for atherosclerotic cardiovascular disease (ACVD), and has also been associated with periodontitis. Inter-individual variation for hsCRP in periodontitis has been shown. ANRIL is the strongest genetic susceptibility locus for both periodontitis and ACVD, and it is speculated that genetic variation in ANRIL may modulate inflammatory processes. Therefore, we explored the possible association between hsCRP plasma levels and a leading ANRIL single nucleotide polymorphism (SNP) in periodontitis patients and controls. 171 healthy subjects with North European descent (115 periodontitis and 56 controls) were included in this case-control study. hsCRP levels were determined and subjects were genotyped for the leading ANRIL SNP rs1333048. In a multivariate analysis, periodontitis, female gender, increasing BMI and homozygosity for the major allele (AA-genotype) of rs1333048 were significantly associated with elevated hsCRP plasma levels (p = 0.012, p = 0.004, p = 0.007 and p = 0.003, respectively). Periodontitis patients with rs1333048 AA-genotype showed higher levels of hsCRP than those carrying the minor C allele (median: 4.5 mg/L vs. 1.6 mg/L, padjusted = 0.007). This study is the first to show that, in addition to gender and BMI, also a leading SNP in ANRIL is explanatory for inter-individual variation in hsCRP levels in periodontitis patients of North European descent.

Periodontitis is associated with platelet activation.

There is an epidemiological association between periodontitis and cardiovascular disease (CVD). In periodontitis, low grade systemic inflammation and bacteremia occur regularly. Such events may contribute to platelet activation and subsequent pro-coagulant state. This study aimed to investigate platelet activation in periodontitis patients. The study is composed of two parts. In the first part, plasma levels of soluble(s) P-selectin and sCD40 ligand were measured as general markers of platelet activation in periodontitis patients (n=85) and in healthy controls (n=35). In the second part, surface-exposed P-selectin and the ligand-binding conformation of the glycoprotein IIb-IIIa complex (binding of PAC-1 antibody) were determined on individual platelets in whole blood of periodontitis patients (n=18) and controls (n=16). Patients had significantly elevated plasma levels of sP-selectin (P<0.001) and increased binding of PAC-1 on isolated platelets (P=0.033). Platelet activation was more pronounced in the patients with more severe periodontal disease, showing a severity-dependence. The levels of sCD40 ligand and of platelet-bound P-selectin were not increased. Periodontitis is associated with increased platelet activation. Since platelet activation contributes to a pro-coagulant state and constitutes a risk for atherothrombosis, platelet activation in periodontitis may partly explain the epidemiological association between periodontitis and CVD.