ICP-Mass-Spectrometry Ionic Profile of Whole Saliva in Patients with Untreated and Treated Periodontitis.
ABSTRACT: Over the past decade, there has been growing interest in the association between macro and trace minerals in body fluids and systemic diseases related to chronic inflammation and oxidative stress. Due to the paucity of data in the literature on periodontitis, the aim of this cross-sectional study was to assess the relationship between mineral elements in saliva and periodontal status in patients with untreated and treated periodontitis compared to periodontally healthy controls. Salivary samples from 66 nonsmoker healthy patients (20 periodontally healthy, 24 untreated severe periodontitis and 22 treated severe periodontitis) were analyzed by using inductively coupled plasma mass-spectrometry (ICP-MS). Significant increases in copper (Cu), sodium (Na), iron (Fe) and manganese (Mn) concentrations occurred in saliva of severe periodontitis subjects compared to periodontally healthy controls. No differences were detected between healthy controls and treated periodontitis patients apart from levels of zinc (Zn) and lithium (Li) that were found to be increased and reduced, respectively, in periodontitis group. Most subjects were correctly separated by cluster analysis into active periodontitis and periodontally healthy individuals. Treated periodontitis individuals were classified as healthy subjects. Based on these preliminary results, the assessment of salivary concentration of mineral elements might be useful in discriminating periodontal health and disease.
Project description:<h4>Background and objective</h4>The complement system is engaged in inflammatory reactions both in the periodontal pockets and in the periodontium itself, where it can mediate tissue destruction. The aim of this study was, first, to compare salivary levels of the total complement system protein C3 and its split product, fluid-phase C3c in patients with periodontitis and periodontally healthy controls. Next, to determine if C3 and C3c levels had biomarker potential in diagnosing and monitoring periodontitis and its treatment. We hypothesized that salivary levels of total C3 and the split product C3c associated with the severity of periodontitis and reflected decreased inflammatory activity after periodontal treatment.<h4>Methods</h4>At baseline, stimulated saliva samples were collected from patients with periodontitis (n = 18) and periodontally healthy controls (n = 15). Subsequently, non-surgical periodontal treatment was performed in the patients, and saliva sampling from patients was repeated two-, six-, and twelve weeks post-treatment (NCT02913248 at clinicaltrials.gov). The patients were grouped as good and poor responders to treatment according to the achieved reduction in bleeding on probing (BOP). Salivary levels of C3 and C3c were quantified using sandwich ELISA.<h4>Results</h4>Patients with periodontitis had higher baseline levels of both total C3 and the split product C3c in saliva than did periodontally healthy controls (P < .0001). Receiver operating curve (ROC) analyses discriminated patients with periodontitis from controls based on both C3 (AUC (area under curve) = 0.91, P < .001) and C3c levels (AUC = 0.84, P < .001) in saliva. Periodontal treatment improved all clinical parameters (P < .01). Good responders (n = 10) had lower baseline levels of C3c than poor responders (n = 8), (P < .05), and baseline levels of C3c discriminated between good and poor responders (AUC = 0.80, P < .05).<h4>Conclusion</h4>In conclusion, patients with periodontitis had higher salivary levels of C3c, and the C3c levels were predictive of reductions in BOP, that is, the poor responders. This suggests that salivary C3c levels possess potential to serve as a biomarker predicting the clinical response to non-surgical periodontal treatment.
Project description:Background:Periodontitis is associated with increased concentration of inflammatory markers and saliva has been proposed as a non-invasive diagnostic fluid in oral and systemic diseases. The levels of salivary biomarkers, such as cytokines, could potentially be used to distinguish periodontal healthy individuals from subjects with periodontal disease. The purpose of this study was to characterize the salivary levels of two inflammatory biomarkers associated with periodontitis, interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-?), in order to assess whether these cytokines salivary levels could potentially be used to complement periodontitis pregnant women diagnose. Methods:Forty-four pregnant women were distributed into three groups, according to their periodontal status: healthy, mild/moderate periodontitis and severe periodontitis. Unstimulated saliva was collected and analysis of TNF-? and IL-6 salivary levels were performed with Immulite®. Results:Women with periodontitis exhibited significantly higher levels (p = 0.001) of salivary IL-6 and TNF-? compared with the healthy group: 25.1 (±11.2) pg/mL vs. 16.3 (±5.0) pg/mL and 29.7 (±17.2) pg/mL vs. 16.2 (±7.6) pg/mL, approximately 1.5 and 1.8 times more, respectively. Additionally, cytokines were significantly increased (p < 0.05) in severe periodontitis compared to periodontal healthy pregnant women. Conclusions:These results revealed that IL-6 and TNF-? salivary biomarkers provide high discriminatory capacity for distinguishing periodontal disease from periodontal health in pregnant women.
Project description:Purpose:The aims of this study were to examine the salivary microbiota in conditions of periodontal health and disease and to explore microbial changes following nonsurgical periodontal treatment. Methods:Non-stimulated saliva samples were collected from 4 periodontally healthy participants at baseline and from 8 patients with chronic periodontitis at baseline and 3 months following nonsurgical periodontal therapy. The V3 and V4 regions of the 16S rRNA gene from the DNA of saliva samples were amplified and sequenced. The salivary microbial compositions of the healthy participants and patients with periodontitis prior to and following nonsurgical treatment of periodontitis were compared based on the relative abundance of various taxa. Results:On average, 299 operational taxonomic units were identified in each sample. The phylogenetic diversity in patients with periodontitis was higher than that in healthy participants and decreased following treatment. The abundance of the phylum Spirochaetes and the genus Treponema in patients with periodontitis was 143- and 134-fold higher than in the healthy control group, respectively, but decreased significantly following treatment. The species that were overabundant in the saliva of patients with periodontitis included the Peptostreptococcus stomatis group, Porphyromonas gingivalis, the Fusobacterium nucleatum group, Parvimonas micra, Porphyromonas endodontalis, Filifactor alocis, and Tannerella forsythia. The phylum Actinobacteria, the genus Streptococcaceae_uc, and the species Streptococcus salivarius group were more abundant in healthy participants than in those with periodontitis. There was a trend toward a decrease in disease-associated taxa and an increase in health-associated taxa following treatment. Conclusions:Our results revealed differences in the taxa of salivary microbiota between conditions of periodontal health and disease. The taxa found to be associated with health or disease have potential for use as salivary biomarkers for periodontal health or disease.
Project description:The human oral cavity is home to a large and diverse community of viruses that have yet to be characterized in patients with periodontal disease. We recruited and sampled saliva and oral biofilm from a cohort of humans either periodontally healthy or with mild or significant periodontal disease to discern whether there are differences in viral communities that reflect their oral health status. We found communities of viruses inhabiting saliva and the subgingival and supragingival biofilms of each subject that were composed largely of bacteriophage. While there were homologous viruses common to different subjects and biogeographic sites, for most of the subjects, virome compositions were significantly associated with the oral sites from which they were derived. The largest distinctions between virome compositions were found when comparing the subgingival and supragingival biofilms to those of planktonic saliva. Differences in virome composition were significantly associated with oral health status for both subgingival and supragingival biofilm viruses but not for salivary viruses. Among the differences identified in virome compositions was a significant expansion of myoviruses in subgingival biofilm, suggesting that periodontal disease favors lytic phage. We also characterized the bacterial communities in each subject at each biogeographic site by using the V3 hypervariable segment of the 16S rRNA and did not identify distinctions between oral health and disease similar to those found in viral communities. The significantly altered ecology of viruses of oral biofilm in subjects with periodontal disease compared to that of relatively periodontally healthy ones suggests that viruses may serve as useful indicators of oral health status.Little is known about the role or the constituents of viruses as members of the human microbiome. We investigated the composition of human oral viral communities in a group of relatively periodontally healthy subjects or significant periodontitis to determine whether health status may be associated with differences in viruses. We found that most of the viruses present were predators of bacteria. The viruses inhabiting dental plaque were significantly different on the basis of oral health status, while those present in saliva were not. Dental plaque viruses in periodontitis were predicted to be significantly more likely to kill their bacterial hosts than those found in healthy mouths. Because oral diseases such as periodontitis have been shown to have altered bacterial communities, we believe that viruses and their role as drivers of ecosystem diversity are important contributors to the human oral microbiome in health and disease states.
Project description:The objective of this study was to investigate the salivary proteins that are associated with periodontitis in patients with Type 2 diabetes mellitus (T2DM). Volunteers for the study were patients from the Diabetic Unit, University of Malaya Medical Centre, whose periodontal status was determined. The diabetic volunteers were divided into two groups, i.e., patients with periodontitis and those who were periodontally healthy. Saliva samples were collected and treated with 10% TCA/acetone/20 mM DTT to precipitate the proteins, which were then separated using two-dimensional polyacrylamide gel electrophoresis. Gel images were scanned using the GS-800(TM) Calibrated Densitometer. The protein spots were analyzed and expressed in percentage volumes. The percentage volume of each protein spot was subjected to Mann-Whitney statistical analysis using SPSS software and false discovery rate correction. When the expression of the salivary proteins was compared between the T2DM patients with periodontitis with those who were periodontally healthy, seven proteins, including polymeric immunoglobulin receptor, plastin-2, actin related protein 3, leukocyte elastase inhibitor, carbonic anhydrases 6, immunoglobulin J and interleukin-1 receptor antagonist, were found to be differentially expressed (p < 0.01304). This implies that the proteins may have the potential to be used as biomarkers for the prediction of T2DM patients who may be prone to periodontitis.
Project description:This study was undertaken to investigate the OPG profiles in gingival crevicular fluid (GCF), saliva, and gingival tissues of chronic periodontitis (CP) patients in response to open flap debridement (OFD).The study included 30 subjects divided into 2 groups: 20 CP patients and 10 periodontally healthy subjects. Plaque index, gingival index, pocket depth, and clinical attachment level measurements were recorded for all subjects. GCF, salivary, and gingival samples were collected from all 30 subjects at baseline and 3 and 6 month after OFD from the 20 CP patients. GCF and salivary OPG levels were assessed by ELISA assay, while OPG expression in gingival tissues was examined by immunohistochemistry.GCF, salivary and gingival OPG profiles were significantly higher in control subjects compared to CP patients at baseline (P < 0.001). Within CP group, OPG levels in GCF, saliva, and gingival samples showed a significant increase at 3 and 6 months after OFD (P < 0.001) compared to baseline. Although OPG values increased significantly in gingival samples and insignificantly in saliva after 3 months compared to 6 months, yet GCF levels were significantly decreased.OPG might be considered as a diagnostic and prognostic biomarker of periodontal bone destruction. This trial is registered with NCT02160613.
Project description:The estimation of oral microbiome (OM) taxonomic composition in periodontally healthy individuals can often be biased because the clinically periodontally healthy subjects for evaluation can already experience dysbiosis. Usually, they are included just based on the absence of clinical signs of periodontitis. Additionally, the age of subjects is used to be higher to correspond well with tested groups of patients with chronic periodontitis, a disorder typically associated with aging. However, the dysbiosis of the OM precedes the clinical signs of the disease by many months or even years. The absence of periodontal pockets thus does not necessarily mean also good periodontal health and the obtained image of "healthy OM" can be distorted.To overcome this bias, we taxonomically characterized the OM in almost a hundred young students of dentistry with precise oral hygiene and no signs of periodontal disease. We compared the results with the OM composition of older periodontally healthy individuals and also a group of patients with severe periodontitis (aggressive periodontitis according to former classification system). The clustering analysis revealed not only two compact clearly separated clusters corresponding to each state of health, but also a group of samples forming an overlap between both well-pronounced states. Additionally, in the cluster of periodontally healthy samples, few outliers with atypical OM and two major stomatotypes could be distinguished, differing in the prevalence and relative abundance of two main bacterial genera: <i>Streptococcus</i> and <i>Veillonella</i>. We hypothesize that the two stomatotypes could represent the microbial succession from periodontal health to starting dysbiosis. The old and young periodontally healthy subjects do not cluster separately but a trend of the OM in older subjects to periodontitis is visible. Several bacterial genera were identified to be typically more abundant in older periodontally healthy subjects.
Project description:<h4>Objectives</h4>The present systematic review examined the available evidence on distinctive salivary ion profile in periodontitis compared to periodontal health and provided a qualitative assessment of the literature.<h4>Background</h4>Macro and trace elements are essential for cellular physiology, and their changes in biological fluids can be revelatory of an underlying pathological status.<h4>Methods</h4>Data from relevant studies identified from PubMed, Embase, and Scopus databases were retrieved to answer the following PECO question: "In systemically healthy individuals, are there any differences in any salivary macro or trace element concentration between periodontally healthy subjects (H) and patients with periodontitis (P)?" Quality of included studies was rated using a modified version of the QUADOMICS tool. A consistency analysis was performed to identify significantly discriminant chemical elements.<h4>Results</h4>After the screening of 873 titles, 13 studies were included reporting data on 22 different elements. Among them, levels of sodium and potassium were consistently and significantly higher in P compared to H. Conflicting results were found for all the other elements, despite concentration of calcium, copper, and manganese mostly increased in saliva of P. Levels of magnesium were found higher in P than in H in 2 studies but lower in 3. Zinc resulted significantly increased in saliva from H compared to P individuals in 2 studies, but one study reported opposite results. Four studies were considered as high quality, while reporting of operative protocols and statistical analysis was a major limitation for the others. Due to high methodologic heterogeneity, meta-analysis was not performed.<h4>Conclusions</h4>Levels of macro or trace elements were differentially identified in saliva across diverse periodontal conditions, having a major potential for investigation of oral homeostasis and for high-resolution periodontal diagnosis. Products of inflammatory physiologic cellular impairment, such as sodium and potassium, were the most consistently associated with periodontitis (PROSPERO CRD42021235744).
Project description:Objective: To profile the salivary microbiomes of a Hong Kong Chinese cohort at a species-level resolution and determine species that discriminated clinically resolved periodontitis from periodontally healthy cases. Methods: Salivary microbiomes of 35 Hong Kong Chinese subjects' under routine supportive dental care were analyzed. All subjects had been treated for any dental caries or periodontal disease with all restorative treatment completed at least 1 year ago and had ?3 residual pockets. They were categorized based on a past diagnosis of chronic periodontitis into "healthy" (H) or "periodontitis" (P) categories. Unstimulated whole saliva was collected, genomic DNA was isolated, and high throughput Illumina MiSeq sequencing of 16S rRNA (V3-V4) gene amplicons was performed. The sequences were assigned taxonomy at the species level by using a BLASTN based algorithm that used a combined reference database of HOMD RefSeqV14.51, HOMD RefSeqExtended V1.1 and GreenGeneGold. Species-level OTUs were subjected to downstream analysis in QIIME and R. For P and H group comparisons, community diversity measures were compared, differentially abundant species were determined using DESeq2, and disease indicator species were determined using multi-level pattern analysis within the R package "indicspecies." Results: P subjects were significantly older than H subjects (p = 0.003) but not significantly different in their BOP scores (p = 0.82). No significant differences were noted in alpha diversity measures after adjusting for age, gender, and BOP or in the beta diversity estimates. Four species; Treponema sp. oral taxon 237, TM7 sp. Oral Taxon A56, Prevotella sp. oral taxon 314, Prevotella sp. oral taxon 304, and Capnocytophaga leadbetteri were significantly more abundant in P than in the H group. Indicator species analysis showed 7 significant indicators species of P group. Fusobacterium sp oral taxon 370 was the sole positive indicator of P group (positive predictive value = 0.9, p = 0.04). Significant indicators of the H category were Leptotrichia buccalis, Corynebacterium matruchotii, Leptotrichia hofstadii, and Streptococcus intermedius. Conclusion: This exploratory study showed salivary microbial species could discriminate treated, well-maintained chronic periodontitis from healthy controls with similar gingival inflammation levels. The findings suggest that certain salivary microbiome features may identify periodontitis-susceptible individuals despite clinical disease resolution.
Project description:It has been shown that bacteria in periodontally diseased patients can be recognized by the detection of volatile metabolites in the headspace of saliva by real-time ambient mass spectrometry. The aim of this study was to use this detection method to analyze the oral metabolome in diseased periodontitis patients before and after therapy to monitor disease evolution and healing events. Twelve patients with advanced chronic periodontal disease and 12 periodontally healthy controls served as test and control groups, respectively. Clinical data, subgingival plaque samples and saliva samples were collected at baseline (BL) and 3 months after treatment. The test group received non-surgical scaling and root planing using systemic antibiotics and the control group received one session of supragingival cleaning. Saliva samples from all subjects were analyzed with ambient mass spectrometry. Significant metabolic alterations were found in the headspace of saliva of periodontitis patients 3 months after the non-surgical periodontal treatment. Furthermore, the diseased group showed metabolic features after the treatment that were similar to the healthy control group. In addition, 29 metabolic features correlated with A. actinomycetemcomitans, 17 features correlated with P. gingivalis and one feature correlated with T. denticola. It was shown that headspace secondary electrospray ionization - mass spectrometry allows the detection of different volatile metabolites in healthy and diseased patients. It can be concluded that this rapid and minimally invasive method could have the potential to routinely diagnose and monitor periodontal diseases in the headspace of saliva samples and, eventually, in exhaled breath.