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AIM To assess the accuracy of self-reported gingival bleeding on brushing (GBoB) for differentiating between periodontal health and disease and explore the optimal haemoglobin concentration that enables visual detection of GBoB. MATERIALS AND METHODS Self-assessment of GBoB was conducted in supervised sessions for 408 consecutive adults. The haemoglobin levels in saliva/toothpaste slurry (TPS) were analysed, followed by a full-mouth periodontal examination. Periodontal diagnoses were made based on the 2017 classification of periodontal diseases. Gingival inflammation was defined as presence of at least 10% of sites with bleeding on probing (BOP). Logistic regression and area under the receiver operating characteristic curve (AUROC) analyses were applied to assess the accuracy of GBoB. RESULTS Overall, 37.1% of the subjects claimed self-reported GBoB, and they had higher values of BOP (median: 25.0%; interquartile range (IQR): 16.0%-37.5%) than those without GBoB (median: 13.5%; IQR: 8.0%-24.8%, p

Aim To assess the diagnostic utility of an oral rinse active matrix metalloproteinase‐8 (aMMP‐8) point‐of‐care test (POCT) for differentiating periodontal health, gingivitis, as well as different stages and grades of periodontitis. Materials & Methods The aMMP‐8 index test was undertaken in 408 consecutive adults, followed by a full‐mouth periodontal examination. The reference standard was the 2017 World Workshop classification of periodontal diseases. Sensitivity, specificity, and the area under the receiver operating characteristic curve (AUROC) were assessed. Results 68.6% of the participants were diagnosed with periodontitis, including Stages I (15.9%), II (15.9%), III (29.7%) and IV (7.1%). A positive aMMP‐8 POCT was associated with periodontitis after adjusting for age, gender, tobacco smoking and systemic diseases, while it was unable to differentiate among the stages/grades of periodontitis and between gingivitis/periodontal health. This test showed a sensitivity of 33.2% and a specificity of 93.0% for detecting periodontitis (threshold level >10 ng/ml). The levels of aMMP‐8 adjusted by the number of teeth present (aMMP‐8/NTP) performed better for periodontitis (sensitivity: 67.1%; specificity: 68.8%). Notably, aMMP‐8/NTP were strongly predictive for Stage IV periodontitis (threshold level =0.4312 ng/ml) (sensitivity: 89.7%; specificity: 73.6%; and AUROC: 0.856). The test performance greatly improved in combination with age and smoking, with a sensitivity of 82.5%, a specificity of 84.4%, and an AUROC of 0.883. Conclusion This aMMP‐8 POCT is able to detect periodontitis with better specificity than sensitivity across the spectrum of its severity. This test may be useful for periodontal screening in conjunction with subject characteristics and/or other sensitive screening tools. Further validation studies are needed.

Aim To clinically validate a self-reported questionnaire for periodontal disease and assess its accuracy for differentiating periodontal health and different stages of periodontitis. Methods A Chinese (Cantonese) version of a self-reported questionnaire was prepared by translating and validating the original English questions proposed by the Centers for Disease Control and Prevention/American Academy of Periodontology (CDC/AAP). The utility of the CDC/AAP questionnaire and its individual questions was assessed against a full-mouth periodontal examination. Periodontal case definition was based on the 2017 World Workshop classification of periodontal diseases. Multivariable logistic regression and the area under the receiver operating characteristic curve (AUROC) analysis were performed to assess the accuracy of the questionnaire. Results 408 subjects enrolled in this study, including those with periodontal health (16.2%), gingivitis (15.2%), Stages I/II periodontitis (31.8%) and Stages III/IV periodontitis (36.8%). Overall, the questionnaire had poor accuracy in detecting the presence of Stages I/II periodontitis with an AUROC 0.608. While it showed moderate to high accuracy for identifying periodontal disease (gingivitis and periodontitis), periodontitis and Stages III/IV periodontitis with an AUROC of 0.837, 0.803 and 0.870, respectively. Self-reported measures in combination with age and tobacco smoking showed excellent performance for identifying Stages III/IV periodontitis with a high AUROC of 0.953, a sensitivity of 95.7%, and a specificity of 89.0%. Specific questions and combinations provided greater utility to discriminate the various periodontal case definitions. Conclusions The self-reported CDC/AAP questionnaire may be a feasible tool for detecting periodontitis, and its combination with demographic and lifestyle factors is useful for the identification of individuals with Stages III/IV periodontitis. This questionnaire seems less helpful in screening of Stages I/II periodontitis. Further studies are needed to test the validity in larger community-based populations.

Background The global burden of periodontal diseases remains high. Population growth trends, changes in risk factors and improved tooth retention will increase the socio-economic burden of periodontitis that is responsible for 3,5 million years lived with disability, 54 billion USD/year in lost productivity and a major portion of the 442 billion USD/year cost for oral diseases. Methods In the context of the Milan World Exhibition 2015 “Feeding the Planet, Energy for Life”, a green paper was developed and offered for global consultation by the European Federation of Periodontology. The final draft was endorsed by professional organizations around the world and is presented to stakeholders as a call for global action. Results Specific actions for the public, policymakers, educators, and professional organizations have been identified in the areas of prevention, detection and care. These actions align public interest and knowledge, need for self-care, professional intervention and policies to the best scientific evidence to proactively promote periodontal health and effectively manage the global burden of periodontal diseases, in accordance with WHO/UN priorities and strategies for tackling common non-communicable diseases (NCDs) via the Common Risk Factor Approach. Conclusions A strong and coherent body of evidence allows identification of actionable preventive, diagnostic and therapeutic strategies to effectively promote periodontal health and general wellbeing, and better manage the socio-economic consequences. Action requires consideration of the specific national scenarios. This article is protected by copyright. All rights reserved.

BACKGROUND Antimicrobial-tolerant microbial persisters critically account for various infections and inflammation. This study identified the characteristics of Porphyromonas gingivalis persisters, and explored their underlying survival mechanisms through proteomic profiling. METHODS Porphyromonas gingivalis cultured with different concentrations of hemin was treated with 100 μg/ml of metronidazole (MTZ). The viability of P. gingivalis persisters was determined by colony-forming unit assay and LIVE/DEAD staining. The proteomic signature of P. gingivalis persister fractions was examined using LC-MS/MS and bioinformatic analysis. RESULTS A small fraction of P. gingivalis persisters survived from lethal MTZ treatment without heritability. At late exponential phase, the frequency of these persisters significantly increased when incubated with 1 μg/ml of hemin compared to 10 μg/ml. Higher levels of P. gingivalis persisters formed at stationary phase than the late exponential phase. High-throughput proteomic analysis showed that the persisters markedly downregulated multiple proteins involved in electron transfer and heme/iron utilization essential for redox regulation and MTZ activation. Moreover, the persisters enabled to shut down major cellular activities (e.g. translation) and overexpress stress proteins. CONCLUSION The presence and survival of metronidazole-tolerant P. gingivalis persisters may be dominated by regulation of cellular redox state and enhanced via repression of heme/iron utilization, dormancy and stress responses.

Background Periodontitis is closely linked with type 2 diabetes mellitus (T2DM) and endothelial dysfunction. This study investigated the effects of periodontal treatment on immuno-inflammatory gene expression of endothelial progenitor cells (EPCs) in diabetic patients. Methods Eighteen T2DM patients with moderate to severe chronic periodontitis were randomly assigned to the Treatment group with oral hygiene instruction (OHI), scaling and root debridement (n = 11), and Control group (n = 7) with OHI alone. Peripheral blood samples were taken for biochemical analysis and culture of EPCs at baseline and 6 months after the treatment. PCR array-based profiling of 84 Toll-like receptor signalling-related genes in EPCs was firstly assessed for four randomly selected patients from the Treatment group. The differentially expressed genes were then further validated by qPCR in both groups. Results All subjects in the Treatment group significantly improved their periodontal conditions. Among the 84 genes tested, IL-6 and IL-8 transcripts were significantly downregulated with over twofold change after the treatment, and this observation was further validated by qPCR in all subjects from both groups (p

Herath TDK, Wang Y, Seneviratne CJ, Lu Q, Darveau RP, Wang CY, Jin L. Porphyromonas gingivalis lipopolysaccharide lipid A heterogeneity differentially modulates the expression of IL-6 and IL-8 in human gingival fibroblasts. J Clin Periodontol 2011; 38: 694–701. doi: 10.1111/j.1600-051X.2011.01741.x. Abstract Aim: Porphyromonas gingivalis lipopolysaccharide (LPS) displays a significant amount of structural heterogeneity, containing both tetra- (LPS1435/1449) and penta-acylated (LPS1690) lipid A structures. This study investigated the effects of the two isoforms of P. gingivalis LPS on the expression of IL-6, IL-8 and TNF-α in human gingival fibroblasts (HGFs). Materials and methods: HGFs were stimulated with P. gingivalis LPS1435/1449 and LPS1690 in both dose- (1 ng–10 μg/ml) and time-dependent (2–48 h) experiments. Total RNA and protein were extracted and used for analysis of the IL-6, IL-8 and TNF-α transcripts as well as IL-6 and IL-8 proteins, by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results: P. gingivalis LPS1690 significantly up-regulated the mRNA and protein expression of IL-6 and IL-8, whereas P. gingivalis LPS1435/1449 did not induce significant host response. The expression levels of IL-6 and IL-8 up-regulated by P. gingivalis LPS1690 continuously increased with time course. In contrast, TNF-α transcript expression was up-regulated promptly by P. gingivalis LPS1690 after 2 h of stimulation and gradually declined afterwards. Conclusions: This study suggests that P. gingivalis LPS heterogeneity may differentially modulate the pro-inflammatory cytokine expression in HGFs, which may contribute to periodontal pathogenesis.

Li X, Tse HF, Yiu KH, Li LSW, Jin L. Effect of periodontal treatment on circulating CD34+ cells and peripheral vascular endothelial function: a randomized controlled trial. J Clin Periodontol 2011; 38: 148–156. doi: 10.1111/j.1600-051X.2010.01651.x. Abstract Aim: Periodontal disease is associated with endothelial dysfunction and increased circulating progenitor cell (CPC) count. This study sought to investigate the effect of periodontal treatment on CPC count and vascular endothelial function. Materials and Methods: A single-blind, randomized controlled trial was conducted in 50 otherwise healthy subjects with moderate-to-severe chronic periodontitis. They were randomly assigned into Treatment group (n=25), in whom periodontal treatment was conducted immediately, and Control group (n=25), in whom periodontal treatment was postponed until the completion of this 3-month study. CPCs and peripheral endothelial function were evaluated at baseline and 3-month follow-up using flow cytometry and peripheral arterial tonometry, respectively. Results: Based on the intention-to-treat analysis, periodontal treatment exhibited neutral effects on endothelial function [treatment effect: 0.03, 95% confidence interval (CI): −0.29 to 0.35, p=0.85]. However, circulating CD34+ cells count significantly decreased in the Treatment group compared with the controls (treatment effect: −29.85 cells/μl, 95% CI: −52.62 to −7.08, p=0.011). The reduction of circulating CD34+ count was positively correlated with the decrease in sites% with bleeding on probing or periodontal pockets 4 mm. Conclusions: This study suggests that treatment of periodontitis has neutral effects on peripheral endothelial function but significantly decreases circulating CD34+ cell count.

Lu Q, Jin L. Human gingiva is another site of C-reactive protein formation. J Clin Periodontol 2010; 37: 789–796. doi: 10.1111/j.1600-051X.2010.01600.x. Abstract Objectives: C-reactive protein (CRP) is primarily synthesized in the liver. It is hypothesized that human gingiva per se may produce CRP and its expression could be associated with IL-6. This study elucidated the CRP expression profile in human gingiva and its possible association with IL-6. Materials and Methods: Ninety-four gingival biopsies were collected from 44 subjects with chronic periodontitis and 18 periodontally healthy subjects. CRP protein was detected by immunohistochemistry and Western blotting, while CRP and IL-6 mRNAs were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR. CRP protein expression in the reconstituted human gingival epithelia (RHGE) was examined by the particle-enhanced immunoturbidimetric assay and Western blotting. Results: CRP protein was detected in gingival tissues from patients and healthy subjects by immunohistochemistry and confirmed by Western blotting. Its expression pattern and level at 16 pairs of periodontal pocket tissues and the adjacent clinically healthy tissues from 16 patients were significantly interrelated (rs=0.693, p

Aim: Emerging evidence shows that periodontal disease is associated with endothelial dysfunction. The purpose of this study was to determine the association between chronic periodontitis (CP) and circulating endothelial progenitor cells (EPC). Materials and Methods: Eighty-six non-smoking subjects (36 males and 50 females, aged 35–80 years) were recruited, including 23 subjects with no or mild CP and 63 subjects with moderate to severe CP. The levels of circulating EPC were quantitatively determined by fluorescence-activated cell analysis, including CD34+/kinase insert domain-containing receptor (KDR)+ (more mature EPC) and CD133+/KDR+ (less mature EPC). Periodontal conditions, the intima–media thickness of carotid arteries and circulating biomarkers were examined. Results: Subjects with moderate to severe CP exhibited an increased risk of high EPC count, compared with those with no or mild CP: CD34+/KDR+ EPC [odds ratio (OR)=9.5, 95% confidence interval (95% CI) 1.5–61.0, p=0.018; CD133+/KDR+ EPC, OR=4.6, 95% CI 1.1–19.5, p=0.039]. C-reactive protein was significantly associated with high CD34+/KDR+ EPC count and age was inversely related with high EPC count. Age, gender and CD34+/KDR+ EPC were independent variables of increased carotid intima–media thickness (p

Objectives: To determine the effect of scaling and root planing (SRP) on the interrelations of subgingival periodontopathogens and both interleukin-8 (IL-8) and granulocyte elastase activity in gingival crevicular fluid (GCF), and to assess their relations to the short-term treatment response in management of chronic periodontitis. Material and methods: GCF and subgingival plaque were collected from 16 subjects with untreated chronic periodontitis at baseline and 4 weeks after SRP. IL-8 levels were determined by ELISA. Granulocyte elastase activity was analyzed with a specific substrate, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA) was calculated. 5 DNA-probes were used to detect the presence of A. actinomycetemcomitans (A. a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.), with a sensitivity = 103 cells/paper point. Results: IL-8 and MR-EA levels in GCF decreased significantly after SRP (p

Porphyromonas gingivalis lipopolysaccharide (LPS) displays a significant amount of structural heterogeneity, containing both tetra- (LPS(1435/1449) ) and penta-acylated (LPS(1690) ) lipid A structures. This study investigated the effects of the two isoforms of P. gingivalis LPS on the expression of IL-6, IL-8 and TNF-α in human gingival fibroblasts (HGFs).HGFs were stimulated with P. gingivalis LPS(1435/1449) and LPS(1690) in both dose- (1 ng-10 μg/ml) and time-dependent (2-48 h) experiments. Total RNA and protein were extracted and used for analysis of the IL-6, IL-8 and TNF-α transcripts as well as IL-6 and IL-8 proteins, by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively.P. gingivalis LPS(1690) significantly up-regulated the mRNA and protein expression of IL-6 and IL-8, whereas P. gingivalis LPS(1435/1449) did not induce significant host response. The expression levels of IL-6 and IL-8 up-regulated by P. gingivalis LPS(1690) continuously increased with time course. In contrast, TNF-α transcript expression was up-regulated promptly by P. gingivalis LPS(1690) after 2 h of stimulation and gradually declined afterwards.This study suggests that P. gingivalis LPS heterogeneity may differentially modulate the pro-inflammatory cytokine expression in HGFs, which may contribute to periodontal pathogenesis.

Aim: Periodontal pathogenesis is characterized by Gram-negative bacteria activation of series of pro- and anti-inflammatory cytokines from host cells through the pathway of lipopolysaccharide (LPS), LPS-binding protein (LBP) and CD14. The present study investigated the expression profiles of interleukin (IL)-1β and IL-10 in periodontal health and disease, and examined the effects of Escherichia coli LPS and LBP interaction on the expression of IL-1β and IL-10 by human gingival fibroblasts (HGF). Material and Methods: Gingival biopsies were collected from 44 subjects with chronic periodontitis and 15 periodontally healthy subjects. The expression of IL-1β and IL-10 was detected by immunohistochemistry. The mRNA expression of IL-1β and IL-10 in HGF was detected by RT-PCR with or without recombinant human LBP (rhLBP), while the peptides were analysed by an enzyme-linked immunosorbent assay. Results: IL-1β was detected in both oral sulcular epithelia of healthy controls and periodontal pocket epithelia of patients. IL-10 was mainly expressed in the intercellular spaces of connective tissues. IL-1β displayed a reverse pattern of expression levels with reference to IL-10, and a negative correlation existed between LBP and the ratio of IL-1β/IL-10. rhLBP suppressed E. coli LPS-induced IL-1β expression by HGF. Conclusion: An appropriate interplay of LBP and cytokines may have a beneficial effect on innate host defence, thereby contributing to periodontal homeostasis.

Aim: To identify risk indicators associated with tooth loss and periodontitis in treated patients responsible for arranging supportive periodontal care (SPC). Materials and Methods: Ninety-seven Chinese subjects (34–77 years) who showed favourable responses to periodontal therapy provided in a teaching hospital 5–12 years previously were recalled. They were advised to seek regular SPC on discharge. Background information, general health status, smoking, oral hygiene habits, follow-up dental care, tooth loss, and periodontal parameters were investigated. Multiple regression analysis was performed. Results: Two hundred and fifty-six teeth had been lost, 195 because of self-reported periodontal reasons. Up to 26.8% sites were with pockets 6 mm. Positive correlations were found between total/periodontal tooth loss and (i) smoking pack-years, (ii) time spent on oral hygiene, (iii) years since therapy's conclusion, (iv) age, and negative correlations with (v) inter-dental brush use, and (vi) education levels. Tooth loss by arch was correlated with wearing of removable partial denture in that arch. Percentage sites with pockets 6 mm were significantly negatively correlated with percentage sites without bleeding on probing. Conclusions: Smokers, more elderly patients, removable partial denture wearers, and patients with lower education levels or not using inter-dental brushes ought to be targeted for clinic-based SPC.

This study aimed to determine the association between the levels of granulocyte elastase and prostaglandin E2 (PGE2) in GCE and the concomitant presence of periodontopathogens in untreated adult periodontitis (AP). GCF and subgingival plaque were sampled by paper strips and paper points respectively, from various periodontal sites in 16 AP subjects. Granulocyte elastase activity in GCF was analyzed with a low molecular weight substrate specific for granulocyte elastase, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA, mAbs/min/site) was calculated. PGE2 levels in GCF were determined by radioimmunoassay. 5 species-specific DNA probes were used to detect the presence of A. actinomyceterncomitans (A.a., ATCC 43718), B. forsythus (B.f, ATCC 43037), P. gingivalis (P.g., ATCC 33277), P. intermedia (P.i., ATCC 33563), and T. denticola (T.d., ATCC 35405), with a sensitivity of 10(3) cells/paper point. No A.a. was detectable from all sites sampled. The predominant combination of species detected was B.f., P.g., P.i. & T.d. and it was significantly higher at periodontitis sites (68%) than at healthy (7%) or gingivitis sites (29%) (p