Identification of Oral Strains of Lactobacillus species in Patients with Chronic Periodontitis and Healthy Individuals

Background and Aims:Recent microbiological researches have revealed the possible role of human cytomegalovirus (HCMV), Epstein barr virus (EBV), and herpes simplex virus (HSV-1 and HSV-2) in the etiopathogenesis of periodontal diseases. The present pilot study has been undertaken to detect the presence of these viruses in chronic periodontitis, aggressive periodontitis, and healthy individuals and to determine the relationship between these viruses and the clinical parameters.Materials and Methods:A total of 10 patients belonging to the age group of 18 to 55 years were included. The patients were randomly assigned into periodontally healthy (sulcus depth ≤ 3 mm), chronic periodontitis, and aggressive periodontitis with pockets measuring ≥6 mm. Seventy-five subgingival plaque samples (25 samples from each group) were collected and subjected to multiplex polymerase chain reaction for the detection of presence of HCMV, EBV, HSV-1, and HSV-2. The results were analyzed using one-way ANOVA for multiple group comparisons followed by Student's t-test for pair-wise comparisons. Categorical data was analyzed by Fisher's exact test.Results:HSV-1 was detected in 76% (P<0.001) of sites with chronic periodontitis and 80% (P<0.001) sites with aggressive periodontitis. EBV was detected in 32% (P<0.05) of sites with chronic periodontitis and aggressive periodontitis. The probing pocket depth and clinical attachment level was statistically significant in HSV-1 detected sites compared with undetected sites in aggressive periodontitis patientsConclusion:Among these viruses HSV-1 and EBV were found to be significantly associated with destructive periodontal disease, including chronic and aggressive periodontitis. Further, HSV-1 was found to be associated with severity and progression of destructive periodontal disease.

Introduction:Periodontitis is a chronic destructive inflammatory disease of the oral cavity. The main causative agent is presence of biofilm formed due to different micro-organisms. Among different micro- organisms “red complex” bacteria is known to be the main causative agent in progression of periodontitis. Porphyromonas gingivalis out of the red the complex organism plays a major role in progression of periodontitis. P. gingivalisis present in both in healthy and diseased individuals. The difference in the strains will determine the virulence factor of the organism and also progression of disease. Only few studies have been done showing variation in strains present between healthy and diseased.Aims:To check the difference in heterogeneity of P. gingivalis in chronic periodontitis and healthy individuals through Arbitrarily Primed-PCR (AP-PCR).Materials and Methods:A total of 400 subjects (200 each of chronic periodontitisandhealthy individuals) were included. Sub-gingival plaque was collected in the Reduced transport fluid (RTF) medium and processed at the institutional central research laboratory. Presence of P. gingivalis was, confirmed by culture andphenotypical analysis. Further confirmed cases were processed for PCR after DNA extraction using 16S rRNA. Positive cases of P. gingivalis were subjected for AP-PCR for clonal analysis using the specific 272 primer.Results:In 152(76%) and 98(49%) were confirmed for P. gingivalis in chronic periodontitis and healthy individual respectively by PCR. AP-PCR analysis showed 6 clusters with similarity index in CP and 3 clusters with similarity index in Healthy individuals.Conclusion:The present study showed difference in clusters between chronic periodontitis and healthy individual'sthussuggestive variantin genetic heterogeneity of P. gingivalis strain between healthy and chronic periodontitis. AP- PCR appears to be a promising tool for clonal analysis of P. gingivalis.

ABSTRACTBackground:Salivary biomarkers can be used to diagnose illnesses, track their course, and gauge a patient’s response to therapy. Inflammatory circumstances result in higher sCD44 levels. The most typical inflammatory disorder connected to periodontal tissue is called periodontitis. Therefore, the purpose “of the current study was to compare the levels of sCD44 in patients with chronic periodontitis before and after scaling and root planing.Objectives:1) Determining CD44 concentrations in healthy individuals’ spit. 2) To quantify CD44 levels in the saliva of individuals with persistent periodontitis. The third objective is to compare the levels of CD44 that can be dissolved in saliva between those with healthy gums and those with active periodontitis. 4) Assessing CD44 levels in saliva after “scaling and root planing in patients with chronic periodontitis.”Materials and Methods:Half of the study participants were classified as having chronic periodontitis, while the other half had healthy gums. Participants with healthy gums and those with chronic periodontitis had their plaque levels, gingival status, and testing depths measured. All the same, data were calculated in persons with chronic periodontitis one month after they had “scaling and root planing done. Clinical attachment levels were evaluated before scaling and root planing was performed on patients with chronic periodontitis and again one month afterwards. Patients with chronic periodontitis,” those whose condition had not improved following scaling and root planing (SRP), and healthy individuals all had spit tests conducted after a month. The concentration of sCD44 in the saliva was determined with a chemically coupled immunosorbent assay.Results:Salivary sCD44 levels were substantially greater in people with chronic periodontitis than in people without the condition. After one month of scaling and root planing in patients with chronic periodontitis, sCD44 levels drastically dropped (p < 0.0001).Conclusion:Chronic periodontitis is linked to elevated levels of salivary sCD44. sCD44 can be employed as a possible salivary biomarker for chronic Periodontitis.

Background: This is the first study showing the co-relationship of apelin among chronic periodontitis (CP) and type 2 diabetes mellitus (T2DM). CP has an anti-inflammatory and anti-glycaemic role in our system. In the future, a localised therapeutic dose of apelin can be used to lower the severity of periodontal disease by reducing the inflammation. The objective of the present study is to investigate the levels of serum apelin from patients with CP, CP + T2DM, and healthy individuals. Materials and Methods: This study included 180 individuals that were equally divided into three groups. Group A consisted of healthy individuals, Group B with CP, and Group C with CP + T2DM. Probing pocket depth (PPD) and serum Apelin (AP) were measured. Serum apelin expression was determined using enzyme-linked immunosorbent assay. Statistical analysis was performed using SPSS software. Results: The highest apelin was in CP + T2DM group (24.0 ± 3.78 ng/dl) followed by CP (15.4 ± 2.01 ng/dl) and healthy individuals (7.6 ± 1.18 ng/dl). There was a significant increase in serum apelin (P ≤ 0.05) in Group C individuals. Similarly, PPD was significantly increased in individuals with CP + T2DM. Conclusion: Expression of significant apelin in patients with CP and T2DM suggests a possible role for these adipokines in inflammation and glucose level regulation. In the future, it may be used as a therapeutic agent in curing periodontitis.

Background: Saliva has been reported as a valuable, non-invasive diagnostic tool in monitoring oral and systemic health. It contains biomarkers of inflammation which facilitates to diagnose periodontal disease. Salivary magnesium has a protective effect from periodontal disease due to its capacity to decrease the effect of bacterial toxins on periodontal tissue and reduce the inflammatory response to oral micro biota with some systemic disease like Type 2 diabetes mellitus (DM) and hypertension. Aim: To evaluate the salivary magnesium levels in Type 2 diabetes mellitus (DM) and hypertensive patients with or without chronic Periodontitis (CP). Methodology: The study population comprises of 75 individuals, aged between 30 to 65 years divided into 5 groups. Group I- Type 2 DM patients with CP, Group II- Hypertensive patients with CP, Group III- Patients with both Type 2 DM and hypertension with CP, Group IV- CP without any systemic diseases, Group V- Healthy individuals without CP and any other systemic disease. Unstimulated whole saliva is collected by means of standardized spitting technique. And the salivary magnesium levels are analysed using colorimetric method. Results: The salivary magnesium levels were increased in CP patients without associated systemic disease (Group 4). A significant decrease was reported in Hypertensive patients with CP (Group 2) and in patients with both Type 2 DM and hypertension with CP (Group 3)

The role of genetic variations in genes related to innate response, as β-defensin-1 (DEFB1), in the context of chronic periodontitis (CP) and diabetes mellitus type 2 (DM2), is still not clear. The present study evaluates the distribution of DEFB1 single nucleotide polymorphisms (SNPs) 5'-untranslated (5'UTR) region and its relation with the CP in DM2 individuals in northeastern Brazilians. Two hundred and eighty individuals participated in the study, being 116 DM2+CP, 95 CP, and 69 healthy individuals. Three known DEFB1 functional SNPs [-52 G>A (rs1799946), -44 C>G (rs1800972), -20 G>A (rs11362)] were genotyped with allele-specific assays. Association was found for the DEFB1 -20 G>A SNP. The G allele, the GA and GG genotypes were significantly (P<0.05) more frequent in the DM2+CP (59.5%, 50%, and 34.5%, respectively) and CP (61%, 44.2%, and 38.9%, respectively) than in healthy individuals (26.8%, 36.2%, and 8.7%, respectively). The GCG and ACG combinations (-52, -44, -20) were significantly more frequent among DM2+CP and CP than in the healthy individuals. The results indicate that genetic variations of DEFB1 gene (SNP-20: G allele and GA and GG genotypes) and the DEFB1 5'UTR haplotypes (GCG and ACG) may be associated with a susceptibility to CP in DM2 individuals as well as CP individuals without DM2.

Background: Saliva has been reported as a valuable, non-invasive diagnostic tool in monitoring oral and systemic health. It contains biomarkers of inflammation which facilitates to diagnose periodontal disease. Salivary magnesium has a protective effect from periodontal disease due to its capacity to decrease the effect of bacterial toxins on periodontal tissue and reduce the inflammatory response to oral micro biota with some systemic disease like Type 2 diabetes mellitus (DM) and hypertension. Aim: To evaluate the salivary magnesium levels in Type 2 diabetes mellitus (DM) and hypertensive patients with or without chronic Periodontitis (CP). Methodology: The study population comprises of 75 individuals, aged between 30 to 65 years divided into 5 groups. Group I- Type 2 DM patients with CP, Group II- Hypertensive patients with CP, Group III- Patients with both Type 2 DM and hypertension with CP, Group IV- CP without any systemic diseases, Group V- Healthy individuals without CP and any other systemic disease. Unstimulated whole saliva is collected by means of standardized spitting technique. And the salivary magnesium levels are analysed using colorimetric method. Results: The salivary magnesium levels were increased in CP patients without associated systemic disease (Group 4). A significant decrease was reported in Hypertensive patients with CP (Group 2) and in patients with both Type 2 DM and hypertension with CP (Group 3)

Prevotella is a Gram-negative anaerobic bacilli. The phenotypic characteristics of the various species of Prevotella are similar, which often makes it difficult in routine differentiation and identification of all the species. The purpose of the study was to detect and compare presence of Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Prevotella loescheii in subgingival plaque samples of chronic periodontitis and healthy individuals. Two hundred and thirty-six subjects were considered consisting of chronic periodontitis (128) and healthy (108) individuals. Subgingival plaque sample was collected in reduced transport fluid and analyzed. DNA extraction and polymerase chain reaction (PCR) were performed for genus Prevotella followed by positive samples were considered for the detection of selected species through multiplex PCR using specific primers. Out of 236 samples, 94.1% were positive for genus Prevotella. Out of 222 cases P. nigrescens showed the highest number of cases positive (59.5%) followed by P. melaninogenica (57.2%), P. intermedia (55.4%), and P. loescheii (40.1%). Species were analyzed individually between chronic periodontitis and healthy, P. intermedia, P. nigrescens, and P. loescheii showed greater positivity in healthy compared to chronic periodontitis. Positivity for P. melaninogenica was high in chronic periodontitis compared to healthy. The number of positive cases for species, when correlated with clinical parameters showed an increase in mean score for all clinical parameters assessed, suggesting the presence of variation in the prevalence of Prevotella species and geographic variation do exist in oral microflora. Findings suggest that they can be normal commensals and opportunistic.

Introduction: This study is the first to examine the connection between apelin levels in individuals suffering from chronic periodontitis (CP) and those with co-existing type II diabetes mellitus (T2DM). Apelin is recognised for its anti-inflammatory and anti-glycaemic properties. Looking ahead, a targeted therapeutic dose of apelin might lower the severity of periodontal disease by reducing inflammation. Our current study plans to measure serum apelin levels across three groups: Patients with CP, those with both CP and T2DM, and healthy individuals. Materials and Methods: A total of 180 subjects took part in this study, divided into three equal groups. Group A included healthy individuals, Group B consisted of patients with CP and Group C comprised those diagnosed with both CP and T2DM. We evaluated probing pocket depth (PPD) and serum apelin (Advanced placement) levels. Serum apelin was precisely measured using an enzyme-linked immunosorbent assay. All statistical analyses were performed with SPSS software. Results: Our findings show a significantly higher serum apelin concentration ( P ≤ 0.05) in the CP + T2DM group (24.0 ± 3.78 ng/dl). Apelin levels decreased progressively in the CP group (15.4 ± 2.01 ng/dl) and were lowest in healthy individuals (7.6 ± 1.18 ng/dl). At the same time, PPD was also significantly increased in individuals with CP + T2DM. Conclusion: Our findings show significant apelin expression in individuals with CP and T2DM, indicating these adipokines play an important role in both inflammation and glucose regulation. This suggests the potential of using apelin as a treatment for periodontitis future.

Because of the potential association between periodontal disease and inflammation, the purpose of the present study is to examine the level of Toll-like receptor 4 (TLR-4), interleukin-18 (IL-18), and uric acid as markers of the inflammatory host response in the plasma and saliva of healthy individuals and patients with periodontitis. In addition, routine biochemical parameters such as fasting glucose, insulin, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, alanine transaminase (ALT), and aspartate transaminase (AST) were measured. The authors also wanted to check whether patients with chronic periodontitis (CP) exhibit different modulations in salivary and/or plasma concentrations of these parameters compared with clinically healthy individuals. Saliva and plasma samples were collected from 40 patients with CP and 20 healthy individuals. TLR-4 and IL-18 measurements were done using commercially available enzyme-linked immunosorbent assay kits. Total, HDL, and LDL cholesterol; triglycerides; fasting glucose; AST; and ALT levels were analyzed on a biochemistry analysis system using specific kits. Non-parametric tests were used for certain parameters in the statistical analyses because the data did not follow Gaussian distribution. Significant differences were observed in plasma and salivary TLR-4 and IL-18 levels, along with clinical measurements such as plaque index and probing depth, in patients with CP (P < 0.001). The plasma level of TLR-4 was found to be increased from 0.99 to 3.28 ng/mL in patients with CP. Salivary TLR-4 levels also showed a slightly higher increase in the diseased state (12.44 to 29.97 ng/mL). A significant increase of ≈ 46% was recorded in the plasma IL-18 level. However, salivary IL-18 levels rose up to > 5-fold in the patients with CP compared with healthy individuals. The level of plasma uric acid was found to be highly significantly increased compared with control individuals. HDL cholesterol and triglyceride also showed significant differences (P < 0.02 and P < 0.03, respectively). Plasma glucose, total cholesterol, LDL cholesterol, and insulin levels did not show any significant difference. There was only a slight increase in plasma AST and ALT levels between diseased and healthy states (22.55 versus 25.50 IU/L and 12.35 versus 15.95 IU/L, respectively). However, salivary AST and ALT levels showed a ≈ 6-fold rise in the patients with CP compared with the healthy individuals. Cross-correlation analysis in the periodontitis disease group showed a significant association of plasma AST, salivary AST, and salivary ALT with uric acid level. Based on this study, the authors believe that TLR-4, IL-18, and uric acid could have a role in the inflammatory pathology of periodontitis. These parameters are suggested to be useful in the prognosis and diagnosis of CP. However, the mechanistic association of these parameters with inflammatory pathology of patients with periodontitis needs to be further elucidated in a higher number of samples.

Visfatin is considered an inflammatory biomarker in periodontal disease (PD). In this meta-analysis, we aimed to evaluate the relationship between Visfatin biomarker level with PD. In this study, Medline, Scopus, Web of Science, and Google Scholar were searched. We included studies that examined visfatin levels in samples from healthy people and periodontal disease until March 2023. The quality of the selected articles was evaluated using the Newcastle-Ottawa assessment scale. Depending on heterogeneity of studies, random-effects or fixed-effect models were used to pool results and report the standardized mean difference (SMD). After screening the retrieved papers, the related data were extracted. A total of 159 studies were identified, and 16 studies were included in the meta-analysis. In 9 studies, the SMD of visfatin level of gingival crevicular fluid (GCF) in patients with chronic periodontitis (CP) and healthy individuals was 4.32 (p<0.001). In 6 studies, the SMD of salivary visfatin level in patients with CP and healthy individuals was 2.95 (p = 0.004). In addition, in five studies, the SMD of serum visfatin level in patients with CP and healthy individuals was 7.87 (p<0.001). Therefore, Visfatin levels in serum, saliva, and GCF of patients with CP were increased in comparison to healthy individuals. Comparison of visfatin levels in saliva of gingivitis patients and healthy individuals showed a significant increase of visfatin in gingivitis patients (SMD:0.57, P = 0.018), but no significant difference was observed in the mean GCF visfatin level of gingivitis patients and healthy individuals (SMD:2.60, P = 0.090). In addition, the results suggested that there is no difference between gingivitis cases compared to CP patients (SMD:3.59, P = 0.217). Visfatin levels in GCF, serum, and saliva have the potential to be used as a diagnostic biomarker of periodontitis.

ABSTRACTIntroduction: In recent decades, biomarkers have been used to predict the progression of chronic periodontitis. One of these biomarkers is alkaline phosphatase (ALP). Due to limitations of the performed studies, this study was performed to determine the amount of salivary ALP and gingival crevicular fluid in patients with chronic periodontitis and healthy individuals.Materials and Methods:Twenty-three patients with severe chronic periodontitis and 23 healthy individuals referred to the Periodontology Department of Ahvaz Jundishapur School of Dentistry were evaluated in this analytical epidemiological study. Salivary ALP and gingival crevicular fluid (GCF) were measured using ALP measuring kit and Hitachi device.Results:Mean (standard deviation) of ALP enzyme was 19.43 (12.5) in GCF of patients with chronic periodontitis and 12 (1.48) in the healthy group, and it was 80.17 (23.9) in the saliva of patients with periodontitis and 24.78 (4.37) units per litre in the healthy group. There was a significant difference in the mean of this enzyme in GCF and saliva of patients with chronic periodontitis and healthy individuals (P < 0.001).Conclusion:The results showed that mean of ALP enzyme is significantly higher in GCV and saliva of patients with chronic periodontitis than in healthy individuals. Therefore, it seems that this parameter can be used as a useful biochemical parameter for the diagnosis of periodontal disease.

ObjectivesThe aim of this cross-sectional observational study was to compare the prevalence of different oral Candida spp. in patients with Type 2 Diabetes and chronic periodontitis in two oral sites: dorsal surface of the tongue and subgingival area. In order to determine subgingival areas as potential reservoirs of yeasts, this study aimed to find differences in the yeasts’ detection between the dorsum of the tongue, as the oral site most commonly inhabited with microorganisms, and subgingival samples. Additionally, potential predictors for the yeasts prevalence were determined.Material and methodsSubjects (N = 146) were divided into four groups: group A- healthy individuals without periodontitis, group B- healthy individuals with chronic periodontitis, group C- Type 2 Diabetes patients with good glycoregulation and Chronic periodontitis and group D- Type 2 Diabetes patients with poor glycoregulation and Chronic periodontitis. Samples were obtained from the tongue by swabbing. Subgingival plaque samples were taken by paper points and periodontal curette. Isolation and identification of different Candida spp. was done using ChromAgar medium. In addition, germ-tube production and carbohydrate assimilation tests were performed.ResultsThe prevalence of Candida spp. was higher in diabetics with poor glycoregulation. The most frequently isolated species was Candida albicans followed by Candida glabrata and Candida tropicalis. In 15.6% of cases, Candida spp. was present in the subgingival area while absent on the tongue. Multivariate regression model showed that HbA1c was Candida spp. predictor for both locations.ConclusionsOur results confirmed that there are Candida spp. carriers among subjects with clinically healthy oral mucosa. Also, this study identified subgingival areas as potential reservoirs of these pathogenic species. Glycoregulation has been recognized as a positive predictor factor of Candida spp.

Chronic periodontitis (CP) is a disease caused by an impaired immune response to oral bacteria and is often found in individuals with type 2 diabetes mellitus (DM2). Dendritic cells are involved in CP and genetic polymorphisms in the DC-SIGN receptor may modulate susceptibility to the disease. The aim of the study was to investigate the distribution of a single nucleotide polymorphism in the DC-SIGN in individuals with DM2 and CP, non-DM2 individuals with CP and healthy controls and its association with CP in a sample of population. 280 individuals (116 with DM2+CP, 95 with CP and 69 healthy controls) were genotyped using real-time PCR with allelespecific probes. Significant differences (p<0.05) were found among the groups with regard to socioepidemiological variables, as well as clinicalepidemiological variables. With regard to allelic and genotypic distribution, the GG genotype was significantly more frequent among the healthy individuals compared to those with DM2+CP, suggesting less susceptibility to DM2+CP (p=0.030). The AG genotype was also associated with a lower bleeding index compared to the AA genotype in healthy individuals (p=0.016). This is the first record of an association between a variant in DC-SIGN and susceptibility to DM2 and CP.

Chronic periodontitis (CP) is the common form of inflammatory oral disease. Matrix metalloproteinases (MMPs) play a pivotal role in the progression of CP by degrading gingival tissue and its remodelling. Here, we conducted a case-control study to investigate a possible association of single-nucleotide polymorphism of MMP genes and their interaction with CP in the Indian population. A total of 357 DNA samples of venous blood was isolated, of which 157 were identified as CP patients and 200 were healthy individuals. Genotyping of six MMP genes (MMP1, MMP3, MMP7, MMP8, MMP12 and MMP13) was done using polymerase chain reaction following Sanger's method of sequencing. Statistical analyses were performed by SPSS v16.0, R package (SNPassoc). Gene-gene interactions were evaluated by MDR 3.0.2. The frequency of 6A allele of MMP3 -11715A-6A gene polymorphisms (36%) and G allele of MMP8 +17G-C gene polymorphisms (34%) were higher in the CP population compared with the healthy population (19% and 24%, respectively). A significant association of T allele of MMP8 -799C-T gene promoter polymorphism was found with CP (OR = 2.95, 95%CI = 2.16 - 4.04, P < 0.0001). Genotypic frequency of MMP12 -82A-G polymorphism is associated with CP risk while its allelic distribution is not (OR = 1.32, 95%CI = 0.93 - 1.88, P = 0.129). Gene-gene interactions show the best cross validation consistency model, i.e. MMP1 -519A-G X MMP7 -181A-G X MMP8 -799C-T polymorphismswith a value of 9/10. This gene-gene interaction shows that the significant association of MMP8 -799C-T polymorphism with CP increased susceptibility. Allelic distribution of MMP8+17G-C and MMP3-11715A-6A polymorphisms revealed their protective role towards decreased risk of CP. MMP1 -519A-G and MMP7 -181A-G polymorphisms show combinatorial synergistic effect on CP risk.