Periodontitis Subjects Research Articles

Association of the Unstimulated Whole Salivary Cytokine IL-1β Levels with Initial, Moderate and Severe Periodontitis. A Case Control Study.

Periodontitis (P) is a highly prevalent inflammatory disease of the oral cavity. The objective of the study was to evaluate the stages of pro-inflammatory cytokine IL-1β in initial, moderate and severe periodontitis. One hundred and twenty two patients were included in the study. Periodontitis subjects had at least 20 natural teeth and ≥8 sites with pocket depths of >4 mm and clinical attachment loss (CAL). A questionnaire was used with respect to the socio demographic parameters which included age, gender, ethnicity, education, marital, residence and occupation. To categorize the severity of the disease, teeth were assessed for, Plaque index (PI), Bleeding on probing (BOP), CAL, missing tooth, tooth mobility and bone loss. Unstimulated whole saliva (UWS) was collected and Interleukin-1β (IL-1β) cytokine levels were analyzed using enzyme linked immunosorbent assay with microplate reader at 450 nm. Clinical parameters and salivary cytokine concentrations were assessed using one-way analysis of variance, whereas a correlation of cases with gender and severity of periodontitis was evaluated using chi-square test. Fifty-nine patients were healthy controls and 63 were periodontitis patients Thirty two percent (n = 20) had initial periodontitis, 40% (n = 25) suffered from moderate and 29% (n = 18) had severe periodontitis. Periodontitis subgroups were significantly different with regards to age and gender (p < 0.001). The mean PPD and CAL among the periodontitis patients (PPD, 3.52 ± 1.25 mm; CAL, 4.04 ± 1.64 mm) were significantly compromised (p < 0.05) compared to healthy controls (PPD, 1.52 ± 0.73 mm; CAL, 0.08 ± 0.28 mm). Increased levels of IL-1β were associated with high CAL and PPD findings. UWS IL-1β levels were higher in periodontitis patients compared to healthy individuals. In addition, cases of severe periodontitis showed significantly higher UWS IL-1β levels compared to initial and moderate periodontitis patients. Comparative levels of salivary IL-1β can be potentially used as a diagnostic tool for periodontitis identification and disease progression along with clinical parameters.

Effect of tumor necrosis factor alpha (TNF-α) -308 and -1031 gene polymorphisms on periodontitis among Saudi subjects

ObjectivesPeriodontitis is an infectious disorder that leads to irreversible loss of the surrounding attachment and bone destruction. Genetic polymorphism of cytokines has been suggested to play a role in periodontitis. This case-control study aimed to investigate the relationship between periodontitis and two single nucleotide polymorphisms (SNPs): rs1800629 (-308 G/A) and rs1799964 (-1031 T/C), in the TNF-α gene promoter area. Materials and methodsPeripheral blood was used to prepare genomic DNA from 60 subjects with stage II to stage III periodontitis, as along with 65 control subjects. Polymerase chain reaction and restriction endonuclease digestion were used to genotype TNF-α SNPs. ResultsThe distribution of both genotypes and alleles of TNF-α (-308 G/A) polymorphism did not vary between periodontitis subjects and the controls (P > 0.05). However, the CT genotype and C allele of the TNF-α (-1031 T/C) polymorphism were observed more frequently in the periodontitis subjects than in the controls, while the TT genotype and the T allele were more predominant in the control subjects than in the periodontitis patients (OR: 3.149; 95% CI: 1.494–6.639; P = 0.002 and OR: 2.933; 95% CI: 1.413–6.090; P = 0.003, sequentially). ConclusionThe TNF-α (-308 G/A) polymorphism potentially has no correlation with periodontitis susceptibility, whereas the TNF-α (-1031) CT genotype and C allele might be related to periodontitis among Saudi subjects.

Candida albicans and Candida dubliniensis in Periodontitis in Adolescents and Young Adults

Aim This study aims to evaluate the association of Candida albicans and Candida dubliniensis with periodontitis in adolescents and young adults in a Moroccan population. Methods 426 subjects aged between 12 and 25 years were recruited for the study. A pool of plaque sample was taken. Samples were cultured on Sabouraud Chloramphenicol medium at 37°C for 24–48 hours and then identified by the Vitek 2 YST system. Clinical data and presence of Candida albicans and Candida dubliniensis were analyzed using Jamovi (Version 1.8). Results Candida albicans was observed in 25 subjects among 68 diseased patients (37%) and in 60 subjects among 358 healthy patients (17%). It can be reported that under normal yeast conditions, there is a statistically significant difference between these two groups (P < 0.001). Candida dubliniensis was more prevalent in periodontitis than in healthy subjects (P=0.026). Regarding clinical variables, subgroups of periodontitis subjects showed significant statistical differences for periodontal probing depth, clinical attachment loss, and number of decayed teeth in advanced periodontitis in comparison with initial or mild periodontitis. The results also indicate that the presence of the two species of Candida is not related to gender or age (P > 0.05) nor related to the severity of the periodontal disease in this population. Conclusion Within the limits of our study, Candida albicans is more frequently associated with periodontitis. The potential role of C. albicans in periodontitis pathogenesis is very complex. More studies on biofilm associated with different forms of periodontitis are necessary. It is also important to assess the coexistence of periodontitis and caries and the associated biofilms.

Effects of Saliva From Periodontally Healthy and Diseased Subjects on Barrier Function and the Inflammatory Response in in vitro Models of the Oral Epithelium.

Background: Periodontitis is a multifactorial, bacteria-mediated chronic inflammatory disease that results in the progressive destruction of the tooth-supporting tissues. It is well-known that saliva from subjects suffering from this disease generally contains higher levels of pro-inflammatory mediators, matrix metalloproteinases (MMP), and bacteria-derived toxic products. The aim of this study was to investigate and compare the effects of saliva from periodontally healthy and diseased subjects on the barrier function and inflammatory response in in vitro models of the oral epithelium.Methods: Unstimulated saliva samples from two groups of subjects, one with a healthy periodontium (n = 12) and one with severe generalized periodontitis (n = 11), were filter-sterilized. All the saliva samples were analyzed using an immunological multiplex assay to determine the levels of various cytokines and MMPs relevant to periodontitis. The impact of saliva on epithelial barrier integrity was assessed by monitoring transepithelial electrical resistance (TER) in an oral epithelium model using the B11 keratinocyte cell line. GMSM-K oral epithelial cells were treated with saliva from both groups to determine their ability to induce the secretion of interleukin-6 (IL-6) and interleukin-8 (IL-8), as determined by an enzyme-linked immunosorbent assay (ELISA).Results: Saliva from the periodontitis subjects contained significantly higher concentrations of matrix metalloproteinase-8 (MMP-8), matrix metalloproteinase-9 (MMP-9), IL-8, and C-X-C motif chemokine ligand 1 (CXCL1) compared to saliva from the healthy subjects. Saliva from the healthy and periodontitis subjects affected cytokine secretion and TER in a similar manner. More specifically, saliva from both groups increased TER and induced IL-6 and IL-8 secretion in the in vitro oral epithelium models used.Conclusion: Independently of the presence or absence of periodontitis, saliva can increase the relative TER and the secretion of IL-6 and IL-8 in in vitro models of the oral epithelium.

Serum Vitamin D level and its association with red blood cell indices in patients with periodontitis.

Research of late has brought to light a connect between Vitamin D and anemia. The level of 25hydroxyvitamin D (25(OH) D) is decreased in periodontitis subjects as against subjects without periodontitis and this reduced level could be related to more risk for anemia in periodontitis patients. This study aimed to evaluate the serum 25(OH) D levels and red blood cell indices in patients affected by generalized Stage III Grade B periodontitis and healthy controls and to assess the association between level of Vitamin D and red cell indices in periodontitis patients. The subjects were categorized into (i) case and (ii) control group of 30 each. Clinical parameters including oral hygiene index simplified, mean ratio of sites that bled on probing, gingival index, probing pocket depth, and the clinical attachment loss were assessed in both the groups. Subjects' blood samples (venous) were taken for the biochemical analysis. In contradiction to healthy subjects, periodontitis subjects had significantly diminished 25(OH) D levels, hemoglobin (Hb), hematocrit, and mean corpuscular hemoglobin concentration (MCHC). 25(OH) D was moderately correlated with MCHC (r = 0.53) and it was statistically significant (P = 0.002). Periodontitis impacts Vitamin D status which further causes anemia. It suggests that effective management of periodontitis can help maintain sufficient Vitamin D status and may be vital in preventing anemia.

Assessment of correlation of COVID-19 infection and periodontitis- A comparative study.

ABSTRACTBackground:Periodontal disease constitutes a group of diseases involving inflammatory aspects of the host caused by several microbial agents that affect periodontal tissues and could have systemic implications.Objective:The present study was conducted to assess the correlation of COVID-19 infection and severity of periodontitis in subjects who has mild form of the disease as compared to subjects having moderate form of the disease.Materials and Methods:The study included 116 subjects suffering from COVID-19 that were equally divided into two groups, each based on a convenient sampling methodology. Group I had a moderate form of COVID that required hospitalization and Group II had a mild form of COVID and were treated at home. The stage of periodontal disease was assessed in both groups. Also, laboratory parameters such as level of C-reactive protein (CRP), white blood cell (WBC), D-dimer, vitamin D, and lymphocytes were also assessed. Statistical analysis was done using Chi-square and multiple logistic regression analysis.Results:More than 75% of subjects in both groups were non-smokers. Subjects having more than one comorbid condition were more in number in Group I (51.7%) as compared to Group II (24%). Severe periodontitis (stages 2–4) was found in 81% of subjects in Group I and 46.2% of subjects in Group II [Figure 1]. The odds of getting severe periodontal disease were 6.32 times more in subjects belonging to Group I as compared to Group II. Subjects having more than one comorbid condition were 4.43 times at risk of severe periodontitis as compared to subjects with no co-morbidityConclusion:Severe form of periodontal disease was associated with moderate-to-severe COVID-19 infection and levels of lymphocytes, WBCs, and CRP were elevated in subjects belonging to Group I.

Comparative evaluation of the levels of nod-like receptor family pyrin domain-containing protein (NLRP) 3 in saliva of subjects with chronic periodontitis and healthy controls.

Context:Periodontitis is an inflammatory disease which is ubiquitous. When there is an onset of infection, the innate immunity gets activated followed by the adaptive immune system. Inflammasomes identify the pathogen-associated molecular patterns or danger-associated molecular patterns and initiate inflammation. Nod- like receptor family pyrin domain-containing protein 3 (NLRP 3) is a protein belonging to the intracellular innate immune sensors that act against bacteria. The inflammasome acts along with the toll-like receptor pathways to initiate an action against pathogens. NLRP3 (also known as PYPAF-1 or cryopyrin) acts via apoptosis-associated speck-like protein (ASC).Aims:The study aimed at finding out the relation between levels of NLRP3 in chronic periodontitis and healthy subjects via the enzyme-linked immunosorbent assay (ELISA).Settings and Design:This was a Cross-sectional study.Materials and Methods:Clinical examination and saliva sampling of the study population was done. Reagents were prepared and NLRP3 levels were estimated using ELISA analysis.Statistical Analysis:Intergroup comparison was initiated using the unpaired t-test and for within the group (intragroup), the two-way analysis of variance was used. The Pearson correlation coefficient helped to determine the strength of linear association.Results:Increased levels of NLRP3 were seen in subjects suffering from chronic periodontitis. NLRP3 was also seen to be positively correlated to probing pocket depth, clinical attachment loss, gingival index, and plaque index.Conclusions:A positive correlation exists between NLRP3 and chronic periodontitis, and hence, NLRP3 can be a potential biomarker.

Evaluation of serum and gingival crevicular fluid levels of Vitamin D binding protein in subjects with clinically healthy periodontium and chronic periodontitis: A clinico bio-chemical study.

Advances in oral and periodontal disease diagnostic research are moving towards methods wherein periodontal risk can be identified and quantified by objective measures such as bio-markers. Given the roles of vitamin D binding protein (DBP) in modulating the immune response and in the transport of vitamin D, it is hypothesised that quantitative changes of vitamin DBP are associated with periodontal disease. The aim of the current study is to measure DBP levels in serum and gingival crevicular fluid (GCF) of patients with generalised chronic periodontitis, in comparison to healthy controls. The present cross-sectional clinico-bio-chemical study includes 30 systemically healthy subjects with 15 periodontally healthy and 15 chronic periodontitis subjects who were recruited from the out-patient Department of Periodontics. GCF and blood samples were collected from all the patients. DBP estimation was performed in both the samples using a commercially available ELISA kit. Serum and GCF DBP levels in chronic periodontitis subjects were significantly higher when compared to the periodontally healthy group. There were no significant correlations found among serum and GCF DBP levels with gender and increasing age in both the groups. An increase in disease severity measured by the increase in probing pocket depth and clinical attachment loss did not show correlation with the GCF and serum DBP levels in the chronic periodontitis group. Based on the findings of the present study, increased serum and GCF DBP levels in chronic periodontitis seem to be a probable marker for identifying ongoing periodontal destruction.

Smoking effect on the interleukin-1β expression and periodontal status of periodontitis patients

Background: The smoking habit is a risk factor for periodontitis. Periodontitis is a multifactorial inflammatory disease associated with biofilm plaque dysbiosis and is characterized by the progressive deterioration of the periodontal tissue. Pro-inflammatory cytokines, such as interleukin-1β (IL-1β), can trigger inflammation of the tissue and become a stimulator to destroy it. Moreover, IL-1β will increase in response to inflammation. Objective: The aim of this study was to evaluate the differences in periodontal status and IL-1β gene expression between smoking and nonsmoking periodontitis subjects. Methods: This research was an analytic observational study with a cross-sectional design. The anthropometric demographic data included name, address, gender, age, plaque index examination, papillary bleeding index (PBI), calculus index, smoking status, and clinical photos to diagnose periodontitis. The collection of gingival crevicular fluid and the analysis of the IL-1β gene expression were conducted using real-time polymerase chain reaction. Results: There were significant differences in periodontal status for plaque index, calculus index, debris index, oral hygiene index, and PBI between smoking and nonsmoking subjects. The IL-1β gene expression was significantly higher (P = 0.047) in smoking subjects than in nonsmoking subjects, with mean ± standard deviation (11.34 ± 9.11) and (0.24 ± 0.42), respectively. Conclusion: This study revealed differences in periodontal status and the IL-1β gene expression between smok ing and nonsmoking periodontitis subjects. The periodontal rate was higher in periodontitis smokers compared to nonsmokers periodontitis. In this study, it was also found that the expression of the IL-1β gene was significantly higher in subjects with smoking periodontitis.

Oxidative stress in human gingival fibroblasts from periodontitis versus healthy counterparts.

Elevated p53 promotes oxidative stress and production of pro-inflammatory cytokines in liposaccharide (LPS)-treated healthy human gingival fibroblasts (HGFs). This study compared oxidative stress, production of inflammatory cytokines, and p53 expression in HGFs from patients with chronic periodontitis (CP) and healthy subjects in vitro upon LPS from Porphyromonas gingivalis challenge. Human gingival fibroblasts were isolated from 6 biopsies-3 from healthy donors and 3 from diseased area in CP (Grade B, Stage III). HGFs were cultured with or without 1μg/ml 24h LPS. Oxidative stress was assessed by analyzing the level of reactive oxygen species (ROS). Mitochondrial membrane potential and respiration were determined by immunofluorescence and respirometry, respectively. Tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β were determined by enzyme-linked immunosorbent assay. P53 expression was monitored by Western blot and immunofluorescence. Human gingival fibroblasts from CP exhibited increased levels of mitochondrial p53, enhanced ROS production, decreased mitochondrial membrane potential, increased mitochondrial oxygen consumption, and increased secretion of TNF-α, IL-6, and IL-1β, as compared to HGFs from healthy donors. Moreover, LPS exacerbated these changes. Human gingival fibroblasts from CP exhibited stronger basal and LPS-inducible oxidative stress and inflammatory response as compared to HGFs from healthy subjects by increased p53 in mitochondria.

Comparison of Some Indicators in Serum and Saliva on Periodontitis

Objectives: To assess the serum indicators level in healthy and periodontitis subjects and correlate the levels in the terms of changes of serum and salivary parameters in periodontal disease. Methods: The assay comprised 85 subjects, enrolled of both genders in the age range of 33-56 years old. Volunteers were categorized into two groups: 40 healthy individual, other groups (periodontal disease) embraced 45 patients. The salivary and blood samples were collected and sent for biochemical analysis to measure the nitric oxide (NO), superoxide dismutase (SOD), glutathione, catalase, Interleukin 6 (IL-6), Interleukin 4 (IL-4), and interferon-γ (IFN-γ) levels in periodontitis patients. Results: Based on the statistical evaluation of bio-indicator parameters, the following conclusion can be shown from this revision: Serum and saliva SOD (P&nbsp;&lt;0.001), glutathione(P&nbsp;&lt;0.001), and catalase(P&nbsp;&lt;0.001) activates were significantly reduced in periodontitis as compared with control groups, the remaining biochemical assessed in periodontitis patients [NO (P&nbsp;&lt;0.001), IL-4 (P&nbsp;&lt;0.0001), IL-6 (P&nbsp;&lt;0.0001), and IFN-γ (P&nbsp;&lt;0.0001)] indicated significantly high levels&nbsp; in both serum and saliva samples. Conclusions: The NO, SOD, catalase, glutathione and cytokines may be used as bio-indicator for periodontitis exposure, medical conduct and severity. This recognition offers early diagnosis of disease and progression. Also there was a direct positive correlation between the salivary and the serum bio-markers levels, which it was proved that the salivary non-invasive examination had a significant association with the serum investigation.

The Varied Proportion of Filifactor alocis in Periodontal Health and Disease in the South Indian Subpopulation.

Background and Aim:The periodontal microbiome being complex, this study was aimed to detect and quantify the prevalence of Filifactor alocis in various stages of periodontitis and to evaluate its prospect as a diagnostic marker for periodontal disease.Settings and Design:Sixty subjects were selected (20 healthy controls, 20 with chronic periodontitis, and 20 with aggressive periodontitis) for the study.Materials and Methods:Clinical parameters probing depth and the level of clinical attachment was recorded, subgingival plaque samples were collected. The F. alocis 16srDNA was cloned, sequenced, and used as the standard for real-time quantification of bacterial load using SYBR green chemistry.Statistical Analysis:Clinical, microbiological, and quantitative polymerase chain reaction (PCR) data were analyzed using ANOVA and Pearson's coefficient correlation.Results:(a) Real-time PCR analysis showed the highest average F. alocis count in chronic periodontitis subjects (32,409.85), which was followed by count in healthy controls (3046.15) and the least count in aggressive periodontitis subjects (939.84). The bacterial count was statistically significant at P = 0.005. (b) An intra-group comparison reveals that there was a statistically significant increase in the bacterial count with age and mean probing pocket depth at P = 0.0005.Conclusion:F. alocis population in aggressive periodontitis was lower compared to chronic periodontitis and healthy controls. The F. alocis population surge in healthy controls may be due to geographical variations and the ethnicity of the subjects. A higher population of F. alocis in chronic periodontitis proves its high pathogenic potential to invade the host tissues to aid in further periodontal destruction.

Antibacterial activity of aquatic extract of Myrtus communis leaves against Periodontitis isolated bacteria

Myrtus communis is an evergreen plant that can survive stressful environments and high-temperature seasons. Treatment using green plants was the most trended in recent years. The present study aimed to evaluate the antibacterial effects of Myrtus communis leaves against bacteria isolated from periodontitis. Fifty samples were collected from periodontitis subjects in both genders (female 32 % and male 68 %). The isolates were diagnosed by morphological characterization and biochemical tests. M. communis leaves were identified, collected, and prepared for extraction. The plant leaves were extracted using distilled water. The antibacterial susceptibility testing was performed by the well diffusion method. Antibiotics susceptibility patterns were executed using the disc diffusion method. All the isolates belonged to gram-positive bacteria. Among the isolated bacteria, 20, 18, and 12 were Lactobacillus spp., Streptococcus spp., and Staphylococcus aureus, respectively. The antibacterial susceptibility testing of M. communis extract showed a potential zone of inhibition against all the tested bacteria. Of the different concentrations, 30 mg/ml showed the highest zone of inhibition, 18.2 mm, 19.50 mm, and 30.66 mm against Streptococcus spp., Staphylococcus spp., and Lactobacillus spp. Among the tested antibiotics, ciprofloxacin and chloramphenicol exhibited the highest zone of inhibition against the tested bacteria. The aquatic extract of M. communis leaves was found to be effective against gram-positive bacteria. Further studies are warranted to investigate the active bio-compounds.

Alterations in subgingival microbiota during full-fixed appliance orthodontic treatment-A prospective study.

Full-fixed appliance orthodontic treatment (commonly called braces) increases plaque accumulation and the risk of gingivitis and periodontitis. However, little consensus exists on changes to subgingival microbiota and specific periodontopathogens during treatment with braces. Prior studies have been hampered by selection biases due to dependence on culture conditions, candidate-based PCR and shallow sequencing methods. The objective was to provide the first longitudinal, culture-free and deep-sequence profiling of subgingival bacteria in subjects during early stages of full-fixed orthodontic treatment. We performed 16S rRNA next-generation sequencing (NGS) on 168 subgingival samples collected at 4 distinct mandibular tooth sites per subject before (0 weeks) and during (6 and 12 weeks) orthodontic intervention in 9 experimental and 5 control subjects not undergoing treatment. Overall, we noted that orthodontic intervention led to increased microbial richness, accompanied by an increased incidence of localized gingivitis/mild periodontitis in subjects requiring orthodontic treatment compared to controls, as well as significant baseline variations in subgingival microbiomes in all subjects. Moreover, we confirmed individual- and site-dependent microbiome variability (in particular, the lingual site harboured higher microbiome diversity than buccal sites) that orthodontic bands may lead to more prolonged shifts in microbial changes compared to brackets, and evidence of adaptive enrichment of consensus bacteria with orthodontic intervention (12 novel, consensus bacterial species were identified). Our study, along with evolving global profiling methods and data analyses, builds a strong foundation for further analyses of subgingival microbiomes during full-fixed orthodontic treatment.

Periodontal disease progression in the second half of life and following a single episode of scaling and root planing: A clinical study in the Sri Lankan tea plantation cohort with documented clinical parameters for more than 40 years.

To evaluate periodontal disease progression (PDP) and potentially detectable effects of a single episode of scaling and root planing (se-SRP) in subjects lacking professional dental care and oral hygiene practices for >40 years. In 2013, se-SRP was offered to all available subjects from the original cohort of 480 males initially established in 1970. From a total of 75 attending the previous examination in 2010 (baseline), 27 consented to receive the intervention while 18 declined and served as controls. Clinical data were recorded again in 2014 (follow-up) similarly to the previous surveys (1970 to 2010). Subjects' mean age in 2010 was 62.5 (± 3.6, test) and 61.9 (± 3.8, control) years. At follow-up, both groups presented with elevated tooth loss of 1.2 (from 15.5 ± 9.0, test) and 1.5 (from 17.9 ± 6.6, control) resulting in 1,392 (test) and 1,061 (control) sites available for further analysis. In both groups, clinical attachment level (CAL) loss and probing depths (PD) deteriorated. PD increase of 0.22mm (± 1.70) in the test group was significantly higher compared with the control group (0.08mm ± 1.30) (P<0.0001) demonstrating unaffected PDP. Computed estimates of further PDP revealed CAL and PD reductions in subjects aged ≥40 years. Specifically, the latter was positively correlated with tooth loss in subjects aged ≥40 years (P=0.69, P=0.0012) and ≥50 years (r=0.62, P<0.0001). se-SRP in previously untreated periodontitis subjects aged ≥50 years may be ineffective in reducing PDP thus demanding advanced preventive measures, treatment in the first half of life, and sustained access to supportive care.

The Expression of Allele Changes in NLRP3 (rs35829419) and IL-1β (+3954) Gene Polymorphisms in Periodontitis and Coronary Artery Disease.

Background: Inflammasomes have been shown to play a pivotal role in periodontal disease pathogenesis. However, their role in periodontitis subjects with coronary heart disease remains unclear. This study aimed to obtain the expression of NLRP3 (rs35829419) and IL-1β (+3954) gene polymorphisms in the subgingival plaque and blood samples of generalized periodontitis (GP) subjects with and without coronary heart disease (CHD). Methods: A total of 70 subjects were grouped into two; GP and GP with CHD. Demographic variables and periodontal and cardiac parameters were recorded from both the groups. Subgingival plaque and blood samples were obtained from both the groups and were further subjected to the identification of NLRP3 (rs35829419) and IL-1β (+3954) expression and allele change using a conventional polymerase chain reaction (PCR) and gene sequencing (Sanger’s method). Results: Amongst the demographic variables, age and monthly income were statistically significant between the two groups. Plaque index (PI), clinical attachment level (CAL), high-density lipoprotein (HDL), and low density-lipoprotein (LDL) exhibited statistically significant levels between the two groups. The NLRP3 (rs35829419) and IL-1β (+3954) genes showed a statistically significant association with allele change (frequency) among the groups. The general comparison of all the parameters with the allele change of NLRP3 (rs35829419) and IL-1β (+3954) in the subgingival plaque and blood samples showed statistically significant associations among the two groups. Conclusion: The present study highlighted an allele change in IL-1β (+3954) gene polymorphisms which may play an important role in the pathogenesis of periodontitis and coronary heart disease.

An Explorative Study on Monocyte Reprogramming in the Context of Periodontitis In Vitro and In Vivo.

AimsPeriodontitis is an independent risk factor for cardiovascular disease, but the mechanistic link is not fully understood. In atherosclerotic cardiovascular disease, monocytes can adopt a persistent hyperresponsive phenotype, termed trained immunity. We hypothesized that periodontitis-associated bacteria can induce trained immunity in monocytes, which subsequently accelerate atherosclerosis development.Materials and MethodsWe combined in vitro experiments on human primary monocytes and in vivo techniques in patients with periodontitis to test this hypothesis. Adherent peripheral blood mononuclear cells (PBMCs) were transiently exposed in vitro to Porphyromonas gingivalis for 24 hours, and restimulated with lipopolysaccharide (LPS) or Pam3CysK4 (P3C) six days later, to measure interleukin-6 (IL-6) and tumor necrosis factor α (TNFα) production. In an exploratory observational study, patients with severe periodontitis (63 ± 6 years, n=14) and control subjects with no-to-mild periodontitis (54 ± 10 years, n=14) underwent venipuncture and 2’-deoxy-2’-[18F]fluoro-D-glucose positron-emission-tomography ([18F]FDG PET/CT) scanning.ResultsWhen adherent peripheral blood mononuclear cells (PBMCs) were transiently exposed in vitro to Porphyromonas gingivalis for 24 hours, and restimulated with LPS or P3C six days later, IL-6 and TNFα production was significantly increased (TNFα/P3C, p<0.01). Circulating leukocytes, IL-6 and interleukin-1 receptor antagonist (IL-1Ra) concentrations were generally higher in patients compared to controls (leukocytes: p<0.01; IL-6: p=0.08; IL-1Ra: p=0.10). Cytokine production capacity in PBMCs after 24h stimulation revealed no differences between groups. [18F]FDG PET/CT imaging showed a trend for increased [18F]FDG-uptake in the periodontium [mean standard uptake value (SUVmean), p=0.11] and in femur bone marrow (SUVmean, p=0.06), but no differences were observed for vascular inflammation. Positive correlations between severity of periodontitis, measured by The Dutch Periodontal Screening Index and pocket depth, with circulating inflammatory markers and tissue inflammation were found.ConclusionsP. gingivalis induces long-term activation of human monocytes in vitro (trained immunity). Patients with severe periodontitis did have signs of increased systemic inflammation and hematopoietic tissue activation. However, their circulating monocytes did not show a hyperresponsive phenotype. Together we suggest that trained immunity might contribute to local periodontal inflammation which warrants further investigation.

Impaired function of epithelial plakophilin-2 is associated with periodontal disease.

Plakophilin-2 (PKP2) is an intracellular desmosomal anchoring protein that has been implicated in a genome-wide association study, in which genetic variants of PKP2 are associated with Porphyromonas gingivalis (P.gingivalis) -dominant periodontal dysbiosis. In this study, we compared the ex vivo PKP2 expression in periodontitis gingival biopsies to periodontitis-free subjects and assessed the in vitro role of PKP2 in gingival epithelial barrier function and the mechanism by which P.gingivalis modulates PKP2 expression. Using reverse transcription quantitative real-time PCR (RT-qPCR), we determined PKP2mRNA expression levels in gingival biopsies collected from 11 periodontally healthy, 10 experimental gingivitis, and 10 chronic periodontitis subjects. PKP2 protein expression in gingival biopsies was detected by immunohistochemistry. We then challenged primary gingival epithelial cells with bacteria including P.gingivalis, Campylobacter rectus, and various Toll-like receptor agonists. Western blot and immunofluorescence staining were used to detect protein expression. Inhibitors blocking proteases pathways were tested for P.gingivalis-mediated PKP2 protein degradations. We also knocked down endogenous epithelial PKP2 using lentiviral short-hairpin RNA (shRNA) and evaluated cell proliferation, spreading, and barrier function. Periodontitis gingival biopsies had approximately twofold less PKP2 mRNA than did healthy controls (p<.05). PKP2 protein was predominantly expressed in gingival epithelium. In primary gingival epithelial cells, P.gingivalis challenge increased PKP2 mRNA levels, while protein expression decreased, which suggests that P.gingivalis has a protein degradation mechanism. Cysteine proteases inhibitors greatly attenuated P.gingivalis-mediated PKP2 protein degradation. Epithelial cells with deficient PKP2 exhibited inhibited cell proliferation and spreading and failed to form monolayers. Finally, P.gingivalis impaired gingival epithelial barrier function. PKP2 appears to be critical in maintaining gingival epithelial barrier function and is susceptible to degradation by cysteine proteases produced by P.gingivalis. Our findings have identified a mechanism by which P.gingivalis impairs epithelial barrier function by promoting PKP2 degradation.

A pilot study of red complex and three genera subgingival microbiome in periodontitis subjects with and without diabetes, evaluated by MinION platform.

Background: Subgingival niche is one biofilm habitat containing rich microbiota, which plays an active role in maintaining the health of periodontal tissue and determining host response. As such, a study of changing subgingival biofilms is important for understanding the effect of a systemic condition. In this study, we compared the occurrence of six bacteria cohabiting in the subgingival area of periodontitis subjects, with (DP, n = 8) and without (NDP, n = 4) diabetes. Methods: The six genus and species of targeted bacteria were confirmed by 16S rRNA amplicon sequencing on MinION nanopore platform. Descriptive statistic was used to describe the obtained data. Results: We found that the six genus and species of targeted bacteria were detected but in different quantities in either group's periodontal pocket. Our data showed that Tannerella forsythia was the most abundant species in subgingival biofilms of the DP group of the red complex bacteria. In contrast, Aggregatibacter sp., which belongs to the phylum of proteobacteria, was present at a relatively lower level. In contrast, Fusobacterium sp., which belongs to orange complex bacteria, showed relative similarities in subgingival biofilms of both groups tested, while Veillonella sp., were abundant in the DP groups. Conclusions: Our data show that the diversity of classic periodontopathogens increased in the subgingival niche of periodontitis subjects with diabetes. It is the first study in Indonesia to apply MinION-based, full-length 16S rRNA amplicon sequencing in periodontitis patients with and without diabetes.

Evaluation of Serum C-Reactive Protein, Plasma Fibrinogen, and Blood Leukocytes in Patients with Chronic Periodontitis

Introduction: Periodontal health is crucial and indispensable element of general health. Epidemiologically, it has been linked with cardiovascular diseases with elevated acute phase reactant C-reactive protein, fibrinogen and rheological variables such as total leukocyte count and differential leukocyte counts, which are potential predictors of cardiovascular diseases.&#x0D; Objective: To evaluate and compare cardiovascular disease-related biochemical markers in periodontally healthy subjects, and patients with moderate and severe chronic periodontitis.&#x0D; Methods: An analytical cross-sectional study was conducted in patients attending Department of Periodontology and Oral Implantology, Chitwan Medical College and Teaching hospital, Bharatpur, Chitwan, Nepal from September 2019 to January 2021. A total of 81 individuals of both gender of which 27 periodontally healthy individuals, 27 diagnosed with moderate and 27 diagnosed with severe periodontitis based on gingival index, probing pocket depth and clinical attachment level were enrolled in a study. After which, peripheral blood samples were drawn and serum C-reactive protein, plasma fibrinogen, total leukocyte count and differential leukocyte counts were quantified using the turbidimetric immunoassay. Convenience sampling technique was done. Data was analysed in SPSS v.26.0.&#x0D; Results: The mean serum levels of C-reactive protein, plasma fibrinogen, total leukocyte and differential leukocytes were found to be statistically (P value &lt;0.001) higher in severe and moderate periodontitis subjects compared to periodontally healthy subjects.&#x0D; Conclusion: The increased levels of serum C-reactive protein, plasma fibrinogen, total leukocyte count, and differential leukocyte counts in chronic periodontitis contribute to the inflammatory burden of the individual potentially striking toward an increasing risk for cardiovascular events.