Estimated economic costs of dental implants among older South Korean adults, 2015-2018.

OBJECTIVE: Dental implants are a considerable financial burden for elderly people and their caregivers. This study aimed to calculate the estimated economic costs of dental implants on the Korean older population. MATERIALS AND METHODS: The economic costs of dental implants for adults aged 75 years and older were estimated from a societal perspective, considering both direct and indirect expenditures. We used data from the Korean National Health Insurance Service for the period 2015-2018 to estimate the medical costs. Noninsured medical costs were estimated from research on medical expenses by the National Health Insurance Review and Assessment Service. Indirect costs related to transportation fees and time loss were obtained. The Cochran Armitage trend test was performed to examine the trend of the economic burden of dental implants. RESULTS: The estimated economic costs showed from 2015 to 2018 (total costs: $26.54-55.66 million, total costs after discount: $17.11-39.56 million). Direct costs, including insured and noninsured medical costs, represented from $25.81 to $47.03 million. Indirect costs, including transportation and time costs, ranged from $0.73 to $1.63 million. The impact of the total dental implant costs was 0.0018%-0.0034% of the Korean annual gross domestic product and 1.03%-1.59% of the annual total costs of dental care benefits. CONCLUSIONS: The estimated economic burden of dental implants significantly increased from 2015 to 2018 in older South Korean adults. These results will provide a foundation and guidance for further health economic studies on the burden of dental implants in the elderly population.

Periodontal tissue susceptibility to glycaemic control in type 2 diabetes.

AIM: To assess the direct effect of intensive glycaemic control on periodontal tissues in patients with diabetes mellitus. MATERIALS AND METHODS: Twenty-nine patients with type 2 diabetes were enrolled and hospitalized to receive a 2-week intensive glycaemic control regimen. We observed and analysed the systemic and oral disease indicators before and after treatment and clarified the indicators related to periodontal inflammation. RESULTS: A significant reduction in glycaemic and periodontal parameters, including glycated albumin levels and periodontal inflamed surface area (PISA), was observed after treatment. The changes in PISA per tooth, indicative of periodontal healing, exhibited a bimodal distribution; the patients were divided into two groups on this basis. Correlations were observed between the changes in PISA per tooth and fasting plasma glucose, acetoacetic acid, and beta-hydroxybutyrate levels in the PISA-improved group. Significantly lower levels of C-peptide, coefficient of variation of R-R interval, and ankle-brachial pressure index were observed before treatment in the PISA non-improved group. CONCLUSIONS: Glycaemic control treatment can effectively improve periodontitis in patients with type 2 diabetes, even in the absence of specific periodontal treatments. However, the periodontal responsiveness to glycaemic control treatment depends on the systemic condition of the patient.

Curcumin inhibits growth of Porphyromonas gingivalis by arrest of bacterial dipeptidyl peptidase activity.

Background: Curcumin is a multi-functional polyphenol with anti-bacterial and anti-inflammatory effects and may have potential for treatment of periodontal diseases. The present study was conducted to examine the molecular basis of the anti-bacterial effect of curcumin against Porphyromonas gingivalis using metabolome analysis. Materials and Methods: P. gingivalis were incubated with 10 µg/mL curcumin, and then metabolites were analyzed with CE-TOF/MS. Expression levels of sigma factors were also evaluated using RT-PCR assays. The activities of dipeptidyl peptidases (DPPs) were assessed by examining the degradation reactions of MCA-labeled peptides. Results: The relative amounts of various glycogenic amino acids were significantly decreased when P. gingivalis was incubated with curcumin. Furthermore, the metabolites on the amino acid degradation pathway, including high-energy compounds such as ATP, various intermediate metabolites of RNA/DNA synthesis, nucleoside sugars and amino sugars were also decreased. Additionally, the expression levels of sigma-54 and sigma-70 were significantly decreased, and the same results as noted following nutrient starvation. Curcumin also significantly suppressed the activities of some DPPs, while the human DPP-4 inhibitors markedly inhibited the growth of P. gingivalis and activities of the DPPs. Conclusions: Curcumin suppresses the growth of P. gingivalis by inhibiting DPPs and also interferes with nucleic acid synthesis and central metabolic pathways, beginning with amino acid metabolism.

Trend in the Incidence of Severe Partial Edentulism among Adults Using the Korean National Health Insurance Service Claim Data, 2014-2018.

PURPOSE: Missing teeth is one of the most important indicators of oral health behavior and the result of dental caries, periodontal disease, and injuries. This study examined a trend in the incidence of severe partial edentulism (SPE) using the Korean National Health Insurance Service (KNHIS) data. MATERIALS AND METHODS: Data of adults aged ≥20 years were obtained from the KNHIS for the 2014-2018 period. SPE was defined in dental information within a population with a treatment history of dental scaling as having 1 to 8 natural teeth. Crude incidence rates (CIRs) and age-standardized incidence rates (AIRs) with 95% confidence interval were calculated per 100000 persons. The Cochran Armitage trend (CAT) test and average annual percentage change were used to analyze SPE trends. RESULTS: The CIRs among Korean adults were from 346.29 to 391.11 in 2014-2016 and from 391.11 to 354.09 in 2016-2018. The AIRs trend statistically increased by 4.31% from 346.29 to 376.80 and decreased by 4.72% from 376.80 to 342.10. The AIRs in men increased by 4.00% and decreased by 3.01%. The AIRs in women decreased by 2.18% and increased by 2.11% (CAT; p<0.01). The AIRs by region and income also showed trends of increase and decrease. CONCLUSION: The study showed that the incidence trend of SPE increased and decreased from 2014 to 2018. This result would be able to aid in the planning of public oral health, and may also serve as fundamental data for verifying the impact of the public oral health policies implemented.

Interspecies metabolite transfer fuels the methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production.

The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from the metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic noncontact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing a new insight into the mechanism of CH3SH generation in oral microbial communities. A better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to the development of more appropriate prophylactic approaches to treat halitosis and periodontitis. An intervention approach like selectively disrupting this interspecies network could also offer a powerful therapeutic strategy.IMPORTANCEHalitosis can have a significant impact on the social life of affected individuals. Among oral odor compounds, CH3SH has a low olfactory threshold and halitosis is a result of its production. Recently, there has been a growing interest in the collective properties of oral polymicrobial communities, regarded as important for the development of oral diseases, which are shaped by physical and metabolic interactions among community participants. However, it has yet to be investigated whether interspecies interactions have an impact on the production of volatile compounds, leading to the development of halitosis. The present findings provide mechanistic insights indicating that ornithine, a metabolite excreted by Streptococcus gordonii, promotes polyamine synthesis by Fusobacterium nucleatum, resulting in a compensatory increase in demand for methionine, which results in elevated methionine pathway activity and CH3SH production. Elucidation of the mechanisms related to CH3SH production is expected to lead to the development of new strategies for managing halitosis.

Cigarette smoke extract impairs gingival epithelial barrier function.

We previously showed that junctional adhesion molecule 1 (JAM1) and coxsackievirus and adenovirus receptor (CXADR), tight junction-associated proteins, have important roles to maintain epithelial barrier function in gingival tissues. Smoking is considered to be a significant risk factor for periodontal disease. The present study was conducted to examine the effects of cigarette smoke extract (CSE) on JAM1 and CXADR in human gingival epithelial cells. CSE was found to cause translocation of JAM1 from the cellular surface to EGFR-positive endosomes, whereas CXADR did not. Using a three-dimensional multilayered gingival epithelial tissue model, CSE administration was found to increase permeability to lipopolysaccharide and peptidoglycan, whereas overexpression of JAM1 in the tissue model prevented penetration by those substrates. Furthermore, vitamin C increased JAM1 expression, and inhibited penetration of LPS and PGN induced by CSE. These findings strongly suggest that CSE disrupts gingival barrier function via dislocation of JAM1, thus allowing bacterial virulence factors to penetrate into subepithelial tissues. Furthermore, they indicate that vitamin C increases JAM1 expression and prevents disruption of gingival barrier function by CSE.

Association of Adiposity with Periodontitis and Metabolic Syndrome: From the Third National Health and Nutrition Examination Survey of United States.

This study explored the epidemiological role of central adiposity and body mass index (BMI) in terms of clinical attachment loss (CAL)/pocket depth (PD) and metabolic syndrome components. This study included data from the National Health and Nutrition Examination Survey III of America on 12,254 adults aged 20 years of age or older with a blood sample, anthropometric measurements, and a periodontal examination. Clinical periodontitis measurements, including CAL and PD, were classified into quintiles or quartiles and compared. CAL was positively associated with central adiposity, hypertension, and hyperglycemia; the relationship between CAL and diabetes was stronger when central adiposity was absent (odds ratio [OR] and 95% confidence interval: 6.33, 2.14-18.72 vs. 3.14, 1.78-5.56). The relationship between CAL and impaired fasting glucose (IFG) differed slightly with BMI. The IFG ORs for normal, overweight, and obese patients were 1.63 (1.08-2.45), 1.76 (1.05-2.97), and 1.43 (0.88-2.30), respectively. CAL was positively correlated with all metabolic syndrome components except hypertriglyceridemia. Associations between CAL, diabetes, and IFG significantly varied with BMI. Periodontitis in individuals without central obesity or with normal bodyweight may independently indicate diabetes and IFG. Therefore, preventive measures against periodontitis without obesity are necessary to improve general and oral health.

Incidence of edentulism among older adults using the Korean National Health Insurance Service database, 2013-2018.

OBJECTIVES: Population aging is rapidly accelerating worldwide. Oral diseases related to aging are also on the rise. This study examined trends in the incidence of edentulism among the older Korean population using data from the Korean National Health Insurance Service (KNHIS). METHODS: Data on older adults, aged ≥75 years of age, were obtained from the KNHIS for the period 2013-2018. Edentulism was defined as a treatment history of complete dentures in the KNHIS database. The exclusion criteria consisted of both disease codes and treatment codes related to conservative dental treatment, including periodontal and extraction treatment afterward. Crude incidence rates (CIRs) and age-standardized incidence rates (AIRs) with 95% confidence intervals were calculated and reported per 100,000 person-years by the direct method. Trends were tested by Cochrane Armitage models. RESULTS: Statistically significant increasing trends in both CIRs and AIRs were found among the older Korean population registered in the KNHIS (CIRs, 707.92 to 895.92; AIRs, 705.11 to 889.68; p<0.01). The incidence tended to increase in both genders (p<0.01). Both CIRs and AIRs in specific regions also showed slight but significant annual increases except for Jeju Island (p<0.01 or <0.05). The incidence showed increasing trends (p<0.01) in all income quintiles apart from the highest quintile. The edentulism incidence was highest in the lowest income group (the first quintile). CONCLUSIONS: Our data showed that the incidence of edentulism among the elderly showed an increasing trend from 2013 to 2018. This result provides a basis for future epidemiological studies on the incidence of edentulism in the older Korean population.

Validity of a combination of periodontal pathogens and salivary biomarkers as predictors of periodontitis.

OBJECTIVE: Chronic periodontitis is caused by multiple risk factors. To predict chronic periodontitis in older people, we evaluated the association between a combination of major periodontal pathogens and salivary biomarkers and the presence of periodontitis. METHODS: Stimulated saliva samples were collected to analyze the prevalence of Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Prevotella intermedia, as well as four biomarkers: interleukin (IL)-1ß, IL-6, tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2). A total of 201 Japanese patients were recruited. Oral examinations ware performed to determine chronic periodontitis as measured by Community Periodontal Index. The sociodemographic and behavioral characteristics were also obtained, and the parameters were adjusted as potential confounders to employ statistical models. RESULTS: The odds ratio (OR) for the presence of P. gingivalis and the third tertile level of IL-1ß as compared with the absence of P. gingivalis and the lowest tertile of IL-1ß was highest in individuals with periodontitis (OR = 13.98; 95% confidence interval [CI] 3.87-50.52) with the best level (0.79) of area under the curve (AUC) based on the receiver operating characteristic curve. The OR for the presence of P. gingivalis and the third tertile of PGE2 was 7.76 (CI 1.89-31.91) with an AUC of 0.78. The coexistence of more than two periodontal bacteria and the third tertile of PGE2 was also strongly associated with chronic periodontitis (OR = 9.23, 95% CI 2.38-35.79) with an AUC of 0.76. CONCLUSIONS: The combined information of the presence of P. gingivalis in stimulated saliva, and higher levels of salivary IL-1ß may play a vital role in the detection and prediction of chronic periodontitis in older adults.

Fusobacterium nucleatum Metabolically Integrates Commensals and Pathogens in Oral Biofilms.

Fusobacterium nucleatum is a common constituent of the oral microbiota in both periodontal health and disease. Previously, we discovered ornithine cross-feeding between F. nucleatum and Streptococcus gordonii, where S. gordonii secretes ornithine via an arginine-ornithine antiporter (ArcD), which in turn supports the growth and biofilm development of F. nucleatum; however, broader metabolic aspects of F. nucleatum within polymicrobial communities and their impact on periodontal pathogenesis have not been addressed. Here, we show that when cocultured with S. gordonii, F. nucleatum increased amino acid availability to enhance the production of butyrate and putrescine, a polyamine produced by ornithine decarboxylation. Coculture with Veillonella parvula, another common inhabitant of the oral microbiota, also increased lysine availability, promoting cadaverine production by F. nucleatum. We confirmed that ArcD-dependent S. gordonii-excreted ornithine induces synergistic putrescine production, and mass spectrometry imaging revealed that this metabolic capability creates a putrescine-rich microenvironment on the surface of F. nucleatum biofilms. We further demonstrated that polyamines caused significant changes in the biofilm phenotype of a periodontal pathogen, Porphyromonas gingivalis, with putrescine accelerating the biofilm life cycle of maturation and dispersal. This phenomenon was also observed with putrescine derived from S. gordonii-F. nucleatum coculture. Lastly, analysis of plaque samples revealed cooccurrence of P. gingivalis with genetic modules for putrescine production by S. gordonii and F. nucleatum. Overall, our results highlight the ability of F. nucleatum to induce synergistic polyamine production within multispecies consortia and provide insight into how the trophic web in oral biofilm ecosystems can eventually shape disease-associated communities. IMPORTANCE Periodontitis is caused by a pathogenic shift in subgingival biofilm ecosystems, which is accompanied by alterations in microbiome composition and function, including changes in the metabolic activity of the biofilm, which comprises multiple commensals and pathogens. While Fusobacterium nucleatum is a common constituent of the supra- and subgingival biofilms, its metabolic integration within polymicrobial communities and the impact on periodontal pathogenesis are poorly understood. Here, we report that amino acids supplied by other commensal bacteria induce polyamine production by F. nucleatum, creating polyamine-rich microenvironments. Polyamines reportedly have diverse functions in bacterial physiology and possible involvement in periodontal pathogenesis. We show that the F. nucleatum-integrated trophic network yielding putrescine from arginine through ornithine accelerates the biofilm life cycle of Porphyromonas gingivalis, a periodontal pathogen, from the planktonic state through biofilm formation to dispersal. This work provides insight into how cooperative metabolism within oral biofilms can tip the balance toward periodontitis.

Surface pre-reacted glass-ionomer eluate protects gingival epithelium from penetration by lipopolysaccharides and peptidoglycans via transcription factor EB pathway.

Surface pre-reacted glass-ionomer (S-PRG) filler, produced by PRG technology for use with various dental materials, is bioactive and known to release ions from a glass-ionomer phase. We previously reported that coxsackievirus and adenovirus receptor (CXADR), a tight junction associated protein, was located in the epithelial barrier of gingival epithelium. In the present study, the tissue protective effects of an S-PRG eluate prepared with S-PRG filler were investigated using a three-dimensional human gingival epithelial tissue model. The results showed that the S-PRG eluate specifically induced CXADR expression at the transcriptional level of messenger RNA as well as the protein level, and also nuclear translocation of transcription factor EB (TFEB) in gingival epithelial cells. Furthermore, shigyakusan, a TFEB inhibitor, canceled induction of the CXADR protein by the S-PRG eluate. Additionally, gingival epithelial permeation by 40-kDa dextran, lipopolysaccharide, and peptidoglycan in the 3D-tissue models was prevented by the eluate, with those effects abrogated by knockdown of CXADR. These findings suggest that S-PRG eluate increases CXADR expression via the TFEB pathway, thus inhibiting penetration of bacterial virulence factors into subepithelial tissues.

Profiling volatile compounds from culture supernatants of periodontal bacteria using gas chromatography/mass spectrometry/olfactometry analysis with a monolithic silica gel adsorption device.

Halitosis is formed mainly by the volatile compounds produced by periodontal bacteria. Three volatile sulfur compounds (VSCs), hydrogen sulfide, methanethiol, and dimethyl sulfide, have attracted attention as major components of halitosis. However, these compounds cannot account for all odors. In this study, we profiled volatile compounds from the culture supernatants of periodontal bacteria using gas chromatography/mass spectrometry/olfactometry analysis with a monolithic silica gel adsorption device to investigate the potential odorous compounds. Periodontal bacteria have been found to produce volatile compounds belonging to various classes, such as alcohols, ketones, fatty acids, and aromatic compounds, in addition to VSCs. In addition, VSCs different from hydrogen sulfide and methanethiol, which are considered important causative compounds, may also influence to halitosis.

Porphyromonas gingivalis Infection Induces Lipopolysaccharide and Peptidoglycan Penetration Through Gingival Epithelium.

Periodontal diseases initiate on epithelial surfaces of the subgingival compartment, while the gingival epithelium functions as an epithelial barrier against microbial infection and orchestrates immune responses. Porphyromonas gingivalis is a major pathogen of periodontal diseases and has an ability to penetrate the epithelial barrier. To assess the molecular basis of gingival epithelial barrier dysfunction associated with P. gingivalis, we newly developed a three-dimensional multilayered tissue model of gingival epithelium with gene manipulation. Using this novel approach, P. gingivalis gingipains including Arg- or Lys-specific cysteine proteases were found to specifically degrade junctional adhesion molecule 1 and coxsackievirus and adenovirus receptor in the tissue model, leading to increased permeability for lipopolysaccharide, peptidoglycan, and gingipains. This review summarizes the strategy used by P. gingivalis to disable the epithelial barrier by disrupting specific junctional adhesion molecules.

Supplemental or dietary intake of omega-3 fatty acids for the treatment of periodontitis: A meta-analysis.

AIM: To evaluate the intervention effect of omega-3 fatty acids on changes in periodontal parameters. MATERIALS AND METHODS: This meta-analysis included studies published in English language between 2010 and 2020, which were extracted from the Cochrane Library, EMBASE, and PubMed databases. The effects of omega-3 fatty acid intervention were investigated using the amount of omega-3 intake, periodontal pocket depth (PPD), clinical attachment loss (CAL), and bleeding on probing (BOP). The random-effects model was generated for data analysis. To obtain robustness of the model, sensitivity analysis was implemented. Subgroup analyses were performed based on the intervention period for each parameter. RESULTS: All 13 studies included in the meta-analysis were interventional, randomized controlled trials. Two studies implemented omega-3 fatty acid-rich diets, while 11 studies used supplements. Risk of bias was low, and publication bias was not shown. Meta-analysis showed a statistically significant PPD reduction (standardized mean difference [SMD] = -0.81, absolute mean difference [MD] = -0.44 mm), CAL gain (SMD = -0.77, MD = -0.51 mm), and BOP reduction (SMD = -0.65, MD = -9.45%) for the omega-3 fatty acid intervention overall. CONCLUSION: This study suggests that supplemental or dietary intake of omega-3 fatty acids for the treatment of periodontitis may have a positive impact on the disease.

Salivary metabolic signatures of carotid atherosclerosis in patients with type 2 diabetes hospitalized for treatment.

Atherosclerosis is a life-threatening disease associated with morbidity and mortality in patients with type 2 diabetes (T2D). This study aimed to characterize a salivary signature of atherosclerosis based on evaluation of carotid intima-media thickness (IMT) to develop a non-invasive predictive tool for diagnosis and disease follow-up. Metabolites in saliva and plasma samples collected at admission and after treatment from 25 T2D patients hospitalized for 2 weeks to undergo medical treatment for diabetes were comprehensively profiled using metabolomic profiling with gas chromatography-mass spectrometry. Orthogonal partial least squares analysis, used to explore the relationships of IMT with clinical markers and plasma and salivary metabolites, showed that the top predictors for IMT included salivary allantoin and 1,5-anhydroglucitol (1,5-AG) at both the baseline examination at admission and after treatment. Furthermore, though treatment induced alterations in salivary levels of allantoin and 1,5-AG, it did not modify the association between IMT and these metabolites (p interaction > 0.05), and models with these metabolites combined yielded satisfactory diagnostic accuracy for the high IMT group even after treatment (area under curve = 0.819). Collectively, this salivary metabolite combination may be useful for non-invasive identification of T2D patients with a higher atherosclerotic burden in clinical settings.

Effect of Molar Occlusal Balance on Cognitive Function in the Elderly.

BACKGROUND: Elderly people with dementia, who are increasing at a rate comparable to the rate at which theelderly population is growing, are becoming a serious social problem in Korea. Therefore, the purpose of this study was to investigate the association between molar occlusal balance and cognitive function among Koreans aged 65 years and older. METHODS: A total of 308 participants aged 65 years and older who attended a senior center were recruited for the study with their consent. The Korean version of the Mini-Mental State Examination (MMSE-DS) was used to assess cognitive function, and masticatory ability was measured according to the ability to chew food, the number of remaining teeth, and the self-perceived perceived masticatory function. Relative molar occlusal balance was measured using the T-scan Ⅲ system. All collected data were analysed using SPSS version 23.0. RESULTS: There was a significant association between cognitive function and molar masticatory ability (P < .05). The participants with relative molar occlusal balance had a higher MMSE-DS score when compared to those with relative incision occlusal balance, adjusted for sociodemographic factors and number of remaining teeth, subjective masticatory ability, chewing ability, occlusion time, and denture use. Cognitive function was higher when relative molar occlusion was greater compared to anterior occlusion in anterior-posterior teeth balance. CONCLUSIONS: Cognitive function in elderly people was higher when the relative molar occlusal balance was greater. Mastication with posterior teeth may have a more important effect on stimulation of cognitive function. Therefore, oral health care focusing on maintenance of molar teeth may be crucial for elderly persons.

Potential of Prebiotic D-Tagatose for Prevention of Oral Disease.

Recent studies have shown phenotypic and metabolic heterogeneity in related species including Streptococcus oralis, a typical oral commensal bacterium, Streptococcus mutans, a cariogenic bacterium, and Streptococcus gordonii, which functions as an accessory pathogen in periodontopathic biofilm. In this study, metabolites characteristically contained in the saliva of individuals with good oral hygiene were determined, after which the effects of an identified prebiotic candidate, D-tagatose, on phenotype, gene expression, and metabolic profiles of those three key bacterial species were investigated. Examinations of the saliva metabolome of 18 systemically healthy volunteers identified salivary D-tagatose as associated with lower dental biofilm abundance in the oral cavity (Spearman's correlation coefficient; r = -0.603, p = 0.008), then the effects of D-tagatose on oral streptococci were analyzed in vitro. In chemically defined medium (CDM) containing D-tagatose as the sole carbohydrate source, S. mutans and S. gordonii each showed negligible biofilm formation, whereas significant biofilms were formed in cultures of S. oralis. Furthermore, even in the presence of glucose, S. mutans and S. gordonii showed growth suppression and decreases in the final viable cell count in a D-tagatose concentration-dependent manner. In contrast, no inhibitory effects of D-tagatose on the growth of S. oralis were observed. To investigate species-specific inhibition by D-tagatose, the metabolomic profiles of D-tagatose-treated S. mutans, S. gordonii, and S. oralis cells were examined. The intracellular amounts of pyruvate-derived amino acids in S. mutans and S. gordonii, but not in S. oralis, such as branched-chain amino acids and alanine, tended to decrease in the presence of D-tagatose. This phenomenon indicates that D-tagatose inhibits growth of those bacteria by affecting glycolysis and its downstream metabolism. In conclusion, the present study provides evidence that D-tagatose is abundant in saliva of individuals with good oral health. Additionally, experimental results demonstrated that D-tagatose selectively inhibits growth of the oral pathogens S. mutans and S. gordonii. In contrast, the oral commensal S. oralis seemed to be negligibly affected, thus highlighting the potential of administration of D-tagatose as an oral prebiotic for its ability to manipulate the metabolism of those targeted oral streptococci.

Saliva and Plasma Reflect Metabolism Altered by Diabetes and Periodontitis.

Periodontitis is an inflammatory disorder caused by disintegration of the balance between the periodontal microbiome and host response. While growing evidence suggests links between periodontitis and various metabolic disorders including type 2 diabetes (T2D), non-alcoholic liver disease, and cardiovascular disease (CVD), which often coexist in individuals with abdominal obesity, factors linking periodontal inflammation to common metabolic alterations remain to be fully elucidated. More detailed characterization of metabolomic profiles associated with multiple oral and cardiometabolic traits may provide better understanding of the complexity of oral-systemic crosstalk and its underlying mechanism. We performed comprehensive profiling of plasma and salivary metabolomes using untargeted gas chromatography/mass spectrometry to investigate multivariate covariation with clinical markers of oral and systemic health in 31 T2D patients with metabolic comorbidities and 30 control subjects. Orthogonal partial least squares (OPLS) results enabled more accurate characterization of associations among 11 oral and 25 systemic clinical outcomes, and 143 salivary and 78 plasma metabolites. In particular, metabolites that reflect cardiometabolic changes were identified in both plasma and saliva, with plasma and salivary ratios of (mannose + allose):1,5-anhydroglucitol achieving areas under the curve of 0.99 and 0.92, respectively, for T2D diagnosis. Additionally, OPLS analysis of periodontal inflamed surface area (PISA) as the numerical response variable revealed shared and unique responses of metabolomic and clinical markers to PISA between healthy and T2D groups. When combined with linear regression models, we found a significant correlation between PISA and multiple metabolites in both groups, including threonate, cadaverine and hydrocinnamate in saliva, as well as lactate and pentadecanoic acid in plasma, of which plasma lactate showed a predominant trend in the healthy group. Unique metabolites associated with PISA in the T2D group included plasma phosphate and salivary malate, while those in the healthy group included plasma gluconate and salivary adenosine. Remarkably, higher PISA was correlated with altered hepatic lipid metabolism in both groups, including higher levels of triglycerides, aspartate aminotransferase and alanine aminotransferase, leading to increased risk of cardiometabolic disease based on a score summarizing levels of CVD-related biomarkers. These findings revealed the potential utility of saliva for evaluating the risk of metabolic disorders without need for a blood test, and provide evidence that disrupted liver lipid metabolism may underlie the link between periodontitis and cardiometabolic disease.

Dynamic analysis of Porphyromonas gingivalis invasion into blood capillaries during the infection process in host tissues using a vascularized three-dimensional human gingival model.

Porphyromonas gingivalis, the pathogen of periodontal disease, is thought to be involved in various diseases throughout the body via gingival tissue blood capillaries. However, the dynamic analysis of the infection mechanism, particularly the deep invasion process of the gingival tissue, has not yet been elucidated because of the lack of both in vivo and in vitro models. In this study, we developed a vascularized three-dimensional (3D) gingival model with an epithelial barrier expressing cell-cell junctions using collagen microfibers (CMFs) to enable the dynamic analysis of the P. gingivalis invasion process. Lipid raft disruption experiments in the gingival epithelial cell layer demonstrated that P. gingivalis migrates into the deeper epithelium via the intercellular pathway rather than intracellular routes. P. gingivalis was shown to invade the 3D gingival model, being found inside blood capillaries during two days of culture. Notably, the number of bacteria had increased greatly at least two days later, whereas the mutant P. gingivalis lacking the cysteine proteases, gingipains, showed a significantly lower number of survivors. The secretion of interleukin-6 (IL-6) from the gingival tissue decreased during the two days of infection with the wild type P. gingivalis, but the opposite was found for the mutant suggesting that P. gingivalis infection disturbs IL-6 secretion at an early stage. By allowing the dynamic observation of the P. gingivalis invasion from the epithelial cell layer into the blood capillaries for the first time, this model will be a powerful tool for the development of novel therapeutics against periodontal infection related diseases.

Porphyromonas gingivalis induces penetration of lipopolysaccharide and peptidoglycan through the gingival epithelium via degradation of coxsackievirus and adenovirus receptor.

Porphyromonas gingivalis is a major pathogen of human periodontitis and dysregulates innate immunity at the gingival epithelial surface. We previously reported that the bacterium specifically degrades junctional adhesion molecule 1 (JAM1), causing gingival epithelial barrier breakdown. However, the functions of other JAM family protein(s) in epithelial barrier dysregulation caused by P. gingivalis are not fully understood. The present results show that gingipains, Arg-specific or Lys-specific cysteine proteases produced by P. gingivalis, specifically degrade coxsackievirus and adenovirus receptor (CXADR), a JAM family protein, at R145 and K235 in gingival epithelial cells. In contrast, a gingipain-deficient P. gingivalis strain was found to be impaired in regard to degradation of CXADR. Furthermore, knockdown of CXADR in artificial gingival epithelium increased permeability to dextran 40 kDa, lipopolysaccharide and peptidoglycan, whereas overexpression of CXADR in a gingival epithelial tissue model prevented penetration by those agents following P. gingivalis infection. Together, these results suggest that P. gingivalis gingipains breach the stratified squamous epithelium barrier by degrading CXADR as well as JAM1, which allows for efficient transfer of bacterial virulence factors into subepithelial tissues. TAKEAWAYS: P. gingivalis, a periodontal pathogen, degraded coxsackievirus and adenovirus receptor (CXADR), a JAM family protein, in gingival epithelial tissues. P. gingivalis gingipains, cysteine proteases, degraded CXADR at R145 and K235. CXADR degradation by P. gingivalis caused increased permeability to lipopolysaccharide and peptidoglycan through gingival epithelial tissues.