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Oral Diseases (2010) 16, 278–285 Aim: To evaluate the salivary carriage of Treponema denticola and its association with demographic variables in the etiopathogenesis of chronic periodontitis. Subjects and methods: Ninety-seven chronic periodontitis (CP) patients and a control group of 51 healthy subjects (HC) were selected. Periodontal status was assessed by criteria based on probing depth, attachment loss, extent, and severity of periodontal breakdown. A polymerase chain reaction method was used to determine the occurrence of T. denticola in saliva samples. Risk indicators for CP were assessed individually and adjusted for confounding and/or interaction using a logistic regression model. Results: Although univariate analysis revealed a positive association of age ≥30 years, smoking, and salivary carriage of T. denticola with CP, after logistic regression analysis, the association between age ≥30 years/smoking and CP persisted, whereas salivary carriage of T. denticola failed to achieve statistical significance. An interaction effect was significantly detected between these three variables. Conclusion: Although salivary carriage of T. denticola may be a risk indicator for CP, its pathogenicity should not be exclusively endorsed to its detection in saliva, but it might be associated with the synergistic biological interaction of the bacterium with some demographic characteristics of the susceptible host. [ABSTRACT FROM AUTHOR]

Aim: To assess the concentration of soluble CD14 receptor in saliva of people with periodontal disease and healthy patients and its relationship with periodontal status. Subjects and Methods: Unstimulated whole saliva samples from patients with chronic periodontitis ( n = 34), aggressive periodontitis ( n = 19) and healthy controls ( n = 17) were obtained for the study. The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss and the extent of periodontal breakdown. The levels of sCD14 were measured in saliva samples with an enzyme-linked immunosorbent assay (ELISA). Results: Although no significant difference ( P > 0.05) was found for salivary sCD14 levels between periodontitis groups, they were significantly greater ( P < 0.05) than those detected for healthy controls. Furthermore, Spearman’s correlation analysis showed statistically significant correlations ( P < 0.01) between data from salivary sCD14 levels and clinical measurements. Conclusion: The findings of the present study re-emphasize the importance of whole saliva as sampling method in terms of immunological purposes in periodontal disease and suggest that the elevated sCD14 concentration may be one of the host-response components associated with the clinical manifestations of periodontal disease. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the in vitro antimicrobial and antibiofilm properties and the immune modulatory activity of cannabidiol (CBD) and cannabigerol (CBG) on oral bacteria and periodontal ligament fibroblasts (PLF)., Methods: Cytotoxicity was assessed by propidium iodide flow cytometry on fibroblasts derived from the periodontal ligament. The minimum inhibitory concentration (MIC) of CBD and CBG for S. mutans and C. albicans and the metabolic activity of a subgingival 33-species biofilm under CBD and CBG treatments were determined. The Quantification of cytokines was performed using the LEGENDplex kit (BioLegend, Ref 740930, San Diego, CA, USA)., Results: CBD-treated cell viability was greater than 95%, and for CBG, it was higher than 88%. MIC for S. mutans with CBD was 20 µM, and 10 µM for CBG. For C. albicans , no inhibitory effect was observed. Multispecies biofilm metabolic activity was reduced by 50.38% with CBD at 125 µg/mL ( p = 0.03) and 39.9% with CBG at 62 µg/mL ( p = 0.023). CBD exposure at 500 µg/mL reduced the metabolic activity of the formed biofilm by 15.41%, but CBG did not have an effect. CBG at 10 µM caused considerable production of anti-inflammatory mediators such as TGF-β and IL-4 at 12 h. CBD at 10 µM to 20 µM produced the highest amount of IFN-γ., Conclusion: Both CBG and CBD inhibit S. mutans ; they also moderately lower the metabolic activity of multispecies biofilms that form; however, CBD had an effect on biofilms that had already developed. This, together with the production of anti-inflammatory mediators and the maintenance of the viability of mammalian cells from the oral cavity, make these substances promising for clinical use and should be taken into account for future studies.

Objective: This work evaluated the Lippia origanoides derivatives in vitro effect on polymicrobial biofilms of Streptococcus mutans, Lactobacillus rhamnosus and Candida albicans. Additionally, the cytotoxic effect of the oils on human skin keratinocytes (HaCaT) and fibroblasts of the periodontal ligament (FLP) cell lines was evaluated., Design: The minimum inhibitory concentration, the inhibitory activity on monomicrobial (S. mutans) and polymicrobial biofilm (S. mutans, L. rhamnosus and C. albicans) of L. origanoides four essential oils and terpenes (thymol and carvacrol) were evaluated. The cytotoxic effect of each one of the compounds was measured, and all the tests were compared against chlorhexidine., Results: All the evaluated compounds reached an inhibition percentage of S. mutans monomicrobial biofilms formation of 100 % at 600 µg/mL (p < 0.0001). The highest concentration (2 MIC) eradicated 100 % of S. mutans-preformed biofilms after 5 min L. origanoides carvacrol + thymol and thymol chemotypes showed marked reductions in topography, the number of microbial cells and extracellular matrix on polymicrobial biofilm. The cytotoxic effect of the compounds was very similar to chlorhexidine., Conclusions: L. origanoides essential oils have an inhibitory effect on mono and polymicrobial biofilms. The oils present a similar cytotoxic effect to chlorhexidine on HaCaT and FLP cell lines. However, including these compounds in formulations for clinical use is an exciting proposal yet to be investigated., Competing Interests: Declaration of Competing Interest The authors present no conflicts of interest to declare. The funding source for this work had no intervention in the design and execution of this research., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Candida albicans infections are related to biofilm formation. The increase in antifungal resistance and their adverse effects have led to the search for therapeutic options as plant derivatives. This scoping review aims to identify the current status of in vitro research on the cytotoxicity and inhibitory effects of plant derivatives on C. albicans biofilms. In this study, PRISMA items were followed. After recognition of the inclusion criteria, full texts were read and disagreements were resolved with a third party. A risk of bias assessment was performed, and information was summarized using Microsoft Office Excel. Thirty-nine papers fulfilling the selection criteria were included. The risk of bias analysis identified most of the studies as low risk. Studies evaluated plant derivatives such as extracts, essential oils, terpenes, alkaloids, flavonoids and polyphenols. Some studies evaluated the inhibition of C. albicans biofilm formation, inhibition on preformed biofilms or both. The derivatives at concentrations greater than or equal to those that have an inhibitory effect on C. albicans biofilms, without showing cytotoxicity, include magnoflorin, ellagic acid, myricetin and eucarobustol from Eucalyptus robusta and, as the works in which these derivatives were studied are of good quality, it is desirable to carry out study in other experimental phases, with methodologies that generate comparable information.

Multi-drug resistant species such as Candida auris are a global health threat. This scenario has highlighted the need to search for antifungal alternatives. Essential oils (EOs), or some of their major compounds, could be a source of new antifungal molecules. The aim of this study was to evaluate the in vitro activity of EOs and some terpenes against C. auris and other Candida spp. The eleven EOs evaluated were obtained by hydro-distillation from different Colombian plants and the terpenes were purchased. EO chemical compositions were obtained by gas chromatography/mass spectrometry (GC/MS). Antifungal activity was evaluated following the CLSI standard M27, 4th Edition. Cytotoxicity was tested on the HaCaT cell line and fungal growth kinetics were tested by time-kill assays. Candida spp. showed different susceptibility to antifungals and the activity of EOs and terpenes was strain-dependent. The Lippia origanoides (thymol + p -cymene) chemotype EO, thymol, carvacrol, and limonene were the most active, mainly against drug-resistant strains. The most active EOs and terpenes were also slightly cytotoxic on the HaCaT cells. The findings of this study suggest that some EOs and commercial terpenes can be a source for the development of new anti- Candida products and aid the identification of new antifungal targets or action mechanisms.

Background: There is a continuing search for compuounds to improve the chemical plaque inhibitory action of oral hygiene products. Although the antibacterial effects of chlorhexidine (CHX) and essential oils components, citral/myrcene, have been described, there is contradictory information regarding their cytotoxic effects in host tissues. This study aimed to evaluate the cytotoxic activity of the major components of the oil C. citratus , citral and myrcene on human periodontal ligament fibroblast (HPLF) cultures and their antimicrobial effect on different bacterial species present in supragingival biofilm., Material and Methods: Cytotoxicity of the compounds to HPLF was determined by MTT assay. Antimicrobial activity was tested against reference strains of Enterococcus faecalis , Streptococcus mutans and Lactobacillus rhamnosus and for S. mutans clinical strains by broth microdilution assay. One-way analysis of variance (ANOVA) with Games-Howell post-hoc multiple comparison or unpaired t tests were used for inter- and intragroup comparisons., Results: Overall, all of the compounds under study showed a cytotoxic effect to HPLF which varied in a dose-dependant manner. Whilst myrcene did not show bacteriostatic activity at tested concentrations, both citral and CHX exhibited bacteriostatic/bactericidal effects to all strains at specific concentrations, being CHX most effective to inhibit bacterial growth at lower concentrations than what observed for citral., Conclusions: Based on these findings, it would possible to conclude that whereas myrcene might be ineffective to control bacterial growth, citral could have a promising antimicrobial activity against dental colonizers with low cytotoxicity, and may be useful for preventing the onset and progression of oral diseases. Key words: Antimicrobial activity, citral, cytotoxicity, chlorhexidine, myrcene., Competing Interests: Conflicts of interest None declared., (Copyright: © 2020 Medicina Oral S.L.)

This study identified potential inhibitory compounds of the phosphoenolpyruvate-sugar. Phosphotransferase system of S. mutans, specifically enzyme II mannose transporter (EII Man ) in its subunits IIA, IIB and IIC by means of a selection protocol and in silico molecular analysis. Intervening the phosphotransferase system would compromise the physiological behavior and the pathogenic expression of S. mutans, and possibly other acidogenic bacteria that use phosphotransferases in their metabolism-making the phosphotransferase system a therapeutic target for the selective control of acidogenic microorganisms in caries control. Several computational techniques were used to evaluate molecular, physicochemical, and toxicological aspects of various compounds. Molecular docking was used to calculate the binding potential (ΔG) between receptor protein subunits and more than 836,000 different chemical compounds from the ZINC database. Physicochemical parameters related to the compounds' pharmacokinetic and pharmacodynamic indicators were evaluated, including absorption, distribution, metabolism, excretion, and toxicity (ADMET), and chemical analysis characterized the compounds structures. Thirteen compounds with EII binding potential of the phosphotransferase system of S. mutans and favorable ADMET properties were identified. Six spirooxindoles and three pyrrolidones stand out from the found compounds; unique structural characteristics of spirooxindoles and pyrrolidones associated with various reported biological activities like anti-microbial, antiinflammatory, anticancer, nootropic, neuroprotective and antiepileptic effects, among other pharmacological effects with surprising differences in terms of mechanisms of action. Following studies will provide more evidence of the action of these compounds on the phosphotransferase system of S. mutans, and its possible applications., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Biofilm is an extremely complex microbial community arranged in a matrix of polysaccharides and attached to a substrate. Its development is crucial in the pathophysiology of oral infections like dental caries, as well as in periodontal, pulp, and periapical diseases. Streptococcus mutans is one of the most effective microorganisms in lactic acid production of the dental biofilm. Identifying essential Streptococcus mutans proteins using bioinformatics methods helps to search for alternative therapies. To this end, the bacterial genomes of several Streptococcus mutans strains and representative strains of other cariogenic and non-cariogenic bacteria were analysed by identifying pathogenicity islands and alignments with other bacteria, and by detecting the exclusive genes of cariogenic species in comparison to the non-pathogenic ones. This study used tools for orthology prediction such as BLAST and OrthoMCL, as well as the server IslandViewer for the detection of pathogenicity islands. In addition, the potential interactome of Streptococcus mutans was rebuilt by comparing it to interologues of other species phylogenetically close to or associated with cariogenicity. This protocol yielded a final list of 20 proteins related to potentially virulent factors that can be used as therapeutic targets in future analyses. The EIIA and EIIC enzymatic subunits of the phosphotransferase system (PTS) were prioritized, as well as the pyruvate kinase enzyme, which are directly involved in the metabolism of carbohydrates and in obtaining the necessary energy for the microorganism's survival. These results will guide a subsequent experimental trial to develop new, safe, and effective molecules in the treatment of dental caries., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Caries is a public health problem, given that it prevails in 60 to 90% of the school-age global population. Multiple factors interact in its etiology, among them dental plaque is necessary to have lactic acid producing microorganisms like Streptococcus from he Mutans group. Existing prevention and treatment measures are not totally effective and generate adverse effects, which is why it is necessary to search for complementary strategies for their management., Aim: The study sought to evaluate the eradication capacity of Streptococcus mutans biofilms and the toxicity on eukaryotic cells of Lippia alba and Cymbopogon citratus essential oils., Methodology: Essential oils were extracted from plant material through steam distillation and then its chemical composition was determined. The MBEC-high-throughput (MBEC-HTP) (Innovotech, Edmonton, Alberta, Canada) assay used to determine the eradication concentration of S. mutans ATCC 35668 strain biofilms. Cytotoxicity was evaluated on CHO cells through the MTT cell proliferation assay., Results: The major components in both oils were Geraniol and Citral; in L. alba 18.9% and 15.9%, respectively, and in C. citratus 31.3% and 26.7%. The L. alba essential oils presented eradication activity against S. mutans biofilms of 95.8% in 0.01mg/dL concentration and C. citratus essential oils showed said eradication activity of 95.4% at 0.1, 0.01mg/dL concentrations and of 93.1% in the 0.001mg/dL concentration; none of the concentrations of both essential oils showed toxicity on CHO cells during 24h., Conclusion: The L. alba and C. citratus essential oils showed eradication activity against S. mutans biofilms and null cytotoxicity, evidencing the need to conduct further studies that can identify their active components and in order to guide a safe use in treating and preventing dental caries., (Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)

Objective: To investigate if the salivary levels of IL-17, IL-21, IL-22, and its ratio regarding salivary IFN-γ may be linked with the periodontal clinical status., Design: One hundred and five chronic periodontitis (CP) subjects and 44 healthy controls (HC) were recruited. Periodontal status was assessed based on full-mouth clinical periodontal measurements. Cytokine salivary levels were analyzed by ELISA. The association between the analytes with CP was analyzed using a binary logistic regression model., Results: A statistically significant increase in salivary levels of IFN-γ and IFN-γ/IL-22 ratio in CP group could be detected, but there was no significant domination of any Th17 cytokine that could be of predictive value for health/disease status. Univariate and binary logistic regression analyses revealed a strong and independent association of IFN-γ salivary levels and IFN-γ/IL-22 ratio with disease status. An interaction effect of ageing on IFN-γ levels also could be noted., Conclusion: While salivary levels of IFN-γ and IFN-γ/IL-22 ratio may act as strong/independent indicators of the amount and extent of periodontal breakdown, the low detection frequency of Th17 cytokines in saliva samples make these determinations useless for the detection of disease presence and/or its severity., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objective: The aim of this study was to compare the effect of two sugar-substituted chewing gums besides toothbrushing on different clinical, microbiological, and biochemical caries- and gingivitis-related variables., Materials and Methods: The study was designed as a double-blind, randomized, controlled trial with three parallel arms. A total of 130 dental students, who volunteered after signing an informed consent, were randomly allocated to receive one of the following interventions: hexitol-sweetened gum containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), pentitol-sweetened gum containing no CPP-ACP, and control group with no gum. Subjects within the experimental groups chewed two gum pellets for 20 min three times a day after meals. The daily consumption level of both polyols was 6.0 g. Clinical examinations and salivary samplings were conducted at baseline and after 30 days of gum use. Pre- and post-intervention stimulated whole saliva samples were quantified for calcium/phosphate ionic concentration, total facultative bacterial load, Streptococcus mutans/Lactobacillus spp. counts, and Gram-negative percentage., Results: A statistically significant reduction in visible plaque score was displayed in the hexitol/CPP-ACP gum group after the intervention when compared with baseline, but the order of the effect was in the same order as the differences between the groups at baseline. A similar tendency was seen in both the pentitol/non-CPP-ACP gum and control groups regarding total salivary facultative bacterial load and S. mutans count, but median values of these parameters were more significantly reduced in the pentitol/non-CPP-ACP gum group in comparison with those of the control group. Alterations of salivary Lactobacillus spp. were demonstrated only in the pentitol/non-CPP-ACP gum group., Conclusion: Although these findings might indicate that a 30-day protocol of daily chewing of pentitol-sweetened gum containing no CPP-ACP might have some a reducing effect on the salivary levels of facultative bacteria, S. mutans and Lactobacillus spp., there was only a marginal, if any, benefit from the chewing gums under study on some microbiological caries- and gingivitis-related variables., Clinical Relevance: Taking into account that for transferring results into clinically relevant conclusions the findings need to be strong and consistent, adhering to single significant differences appears not appropriate. Hence, the clinical significance of chewing gums as an adjunctive tool for daily oral care remained questionable.

Aim: To determine the variations in salivary concentrations of sRANKL, osteoprotegerin (OPG) and its ratio, regarding the periodontal status., Materials and Methods: Ninety-seven chronic periodontitis (CP) subjects and 43 healthy controls were selected. Periodontal status was assessed based on full-mouth clinical periodontal measurements. sRANKL and OPG salivary levels were analysed by ELISA. The association between these analytes and its ratio with CP was analysed individually and adjusted for confounding using a binary logistic regression model., Results: sRANKL and sRANKL/OPG ratio were increased, whereas OPG was decreased in CP compared with healthy controls subjects. Although univariate analysis revealed a positive association of sRANKL salivary levels ≥6 pg/ml, OPG salivary levels ≤131 pg/ml and sRANKL/OPG ratio ≥0.062 with CP, after logistic regression analysis only the latter parameter was strongly and independently associated with disease status. Confounding and interaction effects of ageing and smoking habit on sRANKL and OPG levels could be noted., Conclusion: Although salivary concentrations of sRANKL, OPG and its ratio may act as indicators of the amount/extent of periodontal breakdown, the mutual confounding and synergistic biological interactive effects related to ageing and smoking habit of the susceptible host may also promote the tissue destruction in CP., (© 2012 John Wiley & Sons A/S.)

Objective: Chronic periodontitis (CP) has been linked with an imbalance in the MMP-9/TIMP-1 ratio. A reasonable biologic explanation for this link is that the MMP-9 transcriptional activity can be modulated by MMP-9(-1562C/T) gene promoter polymorphism contributing to periodontal breakdown. This study aimed to assess the relationship between salivary MMP-9/TIMP-1 balance, MMP-9(-1562C/T) genotype and periodontal clinical status., Design: Sixty-nine CP subjects and 54 healthy controls (HC) were selected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, extent, and severity of periodontal breakdown. Salivary levels of MMP-9 and TIMP-1 were analysed using ELISA and MMP-9(-1562C/T) genotype using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The association between salivary levels of MMP-9, TIMP-1 and MMP-9/TIMP-1 ratio with CP was assessed individually and adjusted for confounding using a binary logistic regression model., Results: Significantly higher levels of both markers and their ratios were detected in the CP group in comparison to healthy controls. Synchronously, weak-to-moderate positive significant correlations between salivary biomarkers and clinical parameters were observed. After binary logistic regression analysis, salivary levels of MMP-9>20ngmL(-1), TIMP-1>64ngmL(-1) as well as MMP-9/TIMP-1 ratio >1 were independently associated with CP. Nevertheless, the MMP-9(-1562C/T) gene promoter polymorphism was not associated with the different degrees of chronic periodontitis and did not have influence on the salivary levels of biomarkers., Conclusion: The findings when considered within the limitations of this study may indicate that although a dominant expression of MMP-9 over TIMP-1 in saliva might reflect the periodontal clinical status, the functional polymorphisms in the promoter of the MMP-9(-1562C/T) gene from the Colombian population are not linked neither with significant salivary MMP-9 variations in these individuals nor periodontal clinical status., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Objective: This study aimed to determine whether the variations in the occurrence of gram-negative enteric rods as detected through a longitudinal follow up with three consecutive visits, may be associated with the periodontal clinical status., Design: Clinical and demographic parameters from 63 untreated chronic periodontitis (CP) subjects and 45 healthy controls (HC) were analyzed. Subgingival plaque samples were obtained from 6 sites in each subject at baseline, 1-week, and 1-month visits and processed using culture and biochemical tests. Culture findings were categorized taking into account the detection frequency of gram-negative enteric rods as persistent presence, transient presence, or absence of enteric rods in any sampling time., Results: Although transient presence of gram-negative enteric rods was more prevalent in CP subjects (16.7%) than HC subjects (9.3%), the difference was not significant (P>0.05). The majority of subjects showed a transient presence of gram-negative enteric rods at concentrations <2 x 10(2)CFU/mL. Persistent presence of gram-negative enteric rods was not observed in any subject through the entire study period. Moreover, differences in both the detection frequencies of individual species and the total number of gram-negative enteric rods were not statistically significant neither inter- nor intragroups., Conclusion: The findings of the present study could indicate that gram-negative enteric rods are merely transient microorganisms within the subgingival environment and suggest that the periodontal clinical status appeared not to be influenced by the presence of these species., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Dental caries is a multifactorial disease, in which saliva has an important role. Saliva properties affect the growth of cariogenic microorganisms, so variations below threshold levels are considered risk factors for the development of dental caries. Salivary tests and microbiological counts of cariogenic bacteria are often used for clinical purposes in order to reveal problems that could explain caries development and have been evaluated in multiple studies, but these studies have mainly been carried out on adults. Thus, the aim of this study was to identify physicochemical salivary properties, mutans streptococci and Lactobacillus counts among preschool children, and their relationship with ECC in a cross-sectional study in Medellín, Colombia. Caries was determined using ICDAS criteria for a sample of 201 preschool children. Stimulated saliva samples were processed for bacterial detection, mean flow rate, pH and buffer capacity calculation. Risk variables for ECC were assessed individually and adjusted using a binary logistic regression model. The results showed that 71.14% of study subjects had ECC. Flow rate, final pH and Buffer capacity increased with age, but none of them were related to the presence of ECC. Although OR analysis detected correlations between ECC, gender age, dental eruption stage and Lactobacillus presence, binary logistic regression analysis only showed gender and Lactobacillus as strongly associated with ECC. It can be concluded that gender and Lactobacillus presence are key variables for the presence of ECC in these children; whilst age and dental eruption stage play a minor role. Further studies are required to clarify the role of mutans streptococci counts in this population and to establish normal salivary parameters as well as threshold levels for hyprosalivation in preschool children, as results showed differences with normal parameters used for adults.

The aim of the study was to determine the prevalence of Treponema denticola in saliva of periodontally diseased and healthy patients and its relationship with the periodontal status. A 16S rRNA-based polymerase chain reaction detection method was used to determine the prevalence of T. denticola in whole saliva samples from patients with chronic periodontitis (CP, n = 37), aggressive periodontitis (AgP, n = 24), and healthy subjects (n = 28). The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss, and extent of periodontal breakdown. Risk factors were assessed individually and adjusted for confounding using a binary logistic regression model. The results showed that the prevalence of T. denticola in CP patients was significantly higher than those in healthy and AgP subjects (P < 0.05). Odds ratio analysis revealed a positive association for CP group/T. denticola-positive and smoking/T. denticola-positive subjects. Furthermore, all clinical measurements were significantly greater (P < 0.05) for T. denticola-positive subjects compared to T. denticola-negative subjects. After binary logistic regression analysis, both T. denticola and smoking were independently and strongly associated with development of CP. It was concluded that when used in conjunction with an optimized clinical examination protocol, this assay may offer a rapid, useful, and cost-effective tool for monitoring the presence of T. denticola in noninvasive clinical samples from both healthy and diseased patients and correlating it with the amount and extent of periodontal breakdown.