Pathogen In Periodontitis Research Articles

Novel synthetic peptide derived from Porphyromonas gingivalis Lys-gingipain detects IgG-mediated host response in periodontitis

Porphyromonas gingivalis is a keystone pathogen in periodontitis. Analysis of the immunogenicity of its virulence factors may provide insight into the host response to this infection. The Kgp12 (IEDB Epitope ID 763561), an epitope of Lys-gingipain (Kgp) virulence factor from P. gingivalis ATCC 33277, elicits an immunoglobulin G (IgG) immunoreactivity with low cross-reactivity and, therefore, more specificity. The aim of the present study was to determine in silico the localization of Kgp12 within the protein and to evaluate the IgG host response to this novel Kgp peptide through its capacity to differentiate individuals with different periodontal status. Sera of 71 volunteers were tested by indirect ELISA to detect the IgG immunoreactivity specific to Kgp12, as well as to the protein HmuY and to the sonicated total extract of P. gingivalis ATCC33277, both used as gold standard. The participants had no systemic disease and were classified according to periodontal clinical parameters to comparison, firstly, into periodontitis (P) and without periodontitis (WP) groups and, secondly, into periodontitis (P), gingivitis (G) and clinically health (CH) ones. All the antigens tested, Kgp12 (p = 0.02), HmuY (p = 0.00) and P. gingivalis extract (p = 0.03), could differentiate P from WP groups considering IgG serum levels. P group also had higher IgG levels specific to Kgp12 (p = 0.03), HmuY (p < 0.01) and P. gingivalis extract (p = 0.01) when compared to G group. We conclude that the Kgp12 synthetic peptide was useful to detect the IgG-mediated host response signaling that it is a promising epitope to analyze the immunogenicity of P. gingivalis.

Effect of Sequestosome-1/p62 Expression on Autophagy of Human Periodontal Ligament Fibroblasts Induced by Porphyromonas gingivalis

Porphyromonas gingivalis is the main pathogen of periodontitis. The purpose of this study was to analyze the effect of p62 protein on P. gingivalis-induced autophagy in human periodontal ligament fibroblast (HPDLF). HPDLF cells were first infected with P. gingivalis; autophagy and p62 expression levels were observed at different time points (0, 12, 18, and 24 h) to detect the relationship between p62 and autophagy over time. Autophagy and apoptosis of HPDLF cells infected with P. gingivalis were detected using siRNA specific interference with p62 expression. Fluorescence microscopy and electron microscopy showed autophagic vacuoles in HPDLF cells infected with P. gingivalis. Western blotting showed that p62 expression peaked at 12 h and began to decrease at 18 h. The expression of p62 was silenced by siRNA compared with the control group; the ratio of LC3-II/LC3-I decreased significantly, while the expression level of caspase-3 increased significantly. The the CCK-8 cell viability assay showed that the silencing of p62 reduced HPDLF cell viability, and the plate coating method showed that the silencing of p62 reduced the survival rate of P. gingivalis in the HPDLF cells. Therefore, in vitro studies showed that the presence of p62 is beneficial for maintaining the level of autophagy in HPDLF cells induced by P. gingivalis and inhibiting apoptosis. p62 facilitates the removal of P. gingivalis that are invading cells during this process.

Type IX secretion system is pivotal for expression of gingipain-associated virulence of Porphyromonas gingivalis.

Porphyromonas gingivalis, a keystone pathogen in periodontitis, secretes an array of virulence factors including gingipains via the type IX secretion system (T9SS). Inactivation of any component of the T9SS leads to the accumulation of secreted proteins in unprocessed and, in the case of progingipains, inactive forms in the periplasm. To cast light on the paradox that active gingipains are essential for P.gingivalis fitness in vivo but a functional T9SS is not (Frontiers in Cellular and Infection Microbiology, 2017, 7:378), we have compared virulence of wild-type P.gingivalis W83 and the gingipain-null strain with isogenic mutants deficient in individual T9SS components. Using an in vivo subcutaneous chamber mouse model of infection, gingipain-null strain secretion mutants showed no virulence, but their pathogenic potential was reconstituted by coinfection with a low number of the parental strain. Apparently the same mechanism compensated fitness of mutants lacking functional T9SS the transposon library. In contrast to the parental strain, all mutants elicited significantly lower but an effective inflammatory immune response, which cleared infection and prevented systemic dissemination of P.gingivalis to organs. There were no significant differences in immune responses to different secretion mutants, which were generally more stimulatory than the gingipain-null strain. Together, these results indicate that functional T9SS is essential for P.gingivalis virulence apparently through delivery of active gingipains to the bacterial surface. Therefore, T9SS is a legitimate target for drug development to treat periodontitis.

Celecoxib suppresses lipopolysaccharide-stimulated oral squamous cell carcinoma proliferation in vitro and in vivo.

Periodontitis is one of the most common chronic oral inflammatory conditions worldwide and is associated with a risk of developing oral squamous cell carcinoma (OSCC). Porphyromonas gingivalis is a major pathogen in periodontitis, and its lipopolysaccharide (LPS) promotes the expression of cyclooxygenase-2 (COX-2) in OSCC both in vivo and in vitro. Celecoxib is a selective COX-2 inhibitor; however, its antitumor effects on P. gingivalis LPS-stimulated OSCC and the underlying molecular mechanism remain unclear. To elucidate the association between periodontitis and OSCC, the effect of P. gingivalis-derived LPS on OSCC cell proliferation was examined both in vitro and in vivo in the present study. The expression levels of COX-2 and p53 in OSCC cells with/without celecoxib treatment were determined via western blotting. The therapeutic potential of celecoxib in LPS-stimulated OSCC was evaluated by staining for Ki-67 and p21, as well as with terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling staining. LPS treatment significantly increased OSCC cell proliferation in vitro, and celecoxib significantly inhibited cell proliferation with/without LPS treatment. Celecoxib treatment of OSCC cells downregulated the protein expression levels of COX-2 compared with untreated cells, but there was little change in p53 expression. In the mouse xenograft model, oral administration of celecoxib significantly suppressed tumor growth, reduced the expression of Ki-67, increased the apoptosis index and induced p21 expression with/without LPS treatment. The results from the present study demonstrate that P. gingivalis' LPS can stimulate tumor growth by interacting with OSCC cells. In conclusion, these results suggest that celecoxib could be used for the effective prevention and treatment of LPS-stimulated OSCC.

Research progress on the relationship between Porphyromonas gingivalis and the malignancy of the digestive system and possible pathogenetic mechanism

The malignant tumors including oral cancer, colorectal cancer, pancreatic cancer, and esophageal cancer, of the digestive system are a common high-fatal malignancy. Porphyromonas gingivalis, as the most important pathogen of periodontal disease, has been gradually proved that its invasiveness occurs not only in the mouth but also in other parts of the digestive system. Moreover, the relevant pathogenic mechanism is increasingly attracting the reseachers' attention. In this study, the role and possible pathogenesis of Porphyromonas gingivalis in the digestive system are described in a systematic and comprehensive way.

Microbiological substantiation of application of phytopreparations in treatment of inflammatory periodontal diseases

Relevance:Inflammatory-destructive periodontal diseases are the most complicated and became the main cause of tooth loss in adult population. Herbal medicines have a variety of pharmacological properties, so the development and introduction of new forms for the treatment of inflammatory periodontal diseases is an urgent task today.Purpose– experimental evaluation of effectiveness of the use of herbal medicines “Tonzinal” and “CM-1” in relation to the priority periodontal pathogenes.Materials and methods:in experimental studies, the basis for the experiment was the system for the cultivation of microorganisms in real time – the Revers-Spinner RTS-1 bioreactor. With the priority strains of periodontitis pathogens, the study of the growth dynamics of the culture was carried out in several parallels.Results:herbal medicines “CM-1” and “Tonsinal” has a multilateral therapeutic effect, exerting a diverse influence on the key stages of development of such bacterial populations as Aggregatibacter actinomycetemcomitans, Streptococcus constellatus, Candida albicans.Conclusion:tan integrated approach in the treatment of patients with inflammatory periodontal diseases is promising and will contribute to a more prolonged remission and increase the effectiveness of treatment.

Isolates from Monechma ciliatum seeds' extract hampered Porphyromonas gingivalis hemagglutinins.

Porphyromonas gingivalis is a major periodontitis pathogen that produces several virulence factors including hemagglutinins. These proteins, which are vital molecules, allow P. gingivalis to uptake iron and heme by attaching, aggregating, and lysing erythrocytes. In this study, we evaluated the inhibitory activity of the aqueous extract of Monechma ciliatum seeds against the hemagglutination activity of P. gingivalis. M. ciliatum is a Sudanese medicinal herb that grows in arid and semi-arid lands of tropical Africa. The water extracted from dry powdered seeds was partitioned using ethyl acetate followed by reversed-phase chromatography, thin-layer chromatography, ESI-MS, and NMR analysis resulting in the isolation of four compounds identified as oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol with MICs of 15-100μg/ml against hemagglutination. We believe that the isolation and purification of these compounds will expand the application of M. ciliatum as a natural therapeutic or preventative agent. PRACTICAL APPLICATIONS: Monechma ciliatum or black mahlab is a famous medicinal plant that grows in some parts of arid and semi-arid areas of tropical Africa including western Sudan. Despite its nutritional and traditional medical applications, no studies have evaluated its anti-hemagglutination activity against periodontal pathogens. In this study, four active compounds (oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol) were isolated and identified from an aqueous extract of M. ciliatum seeds. The isolated compounds revealed high levels of inhibitory activity against all hemagglutinin agents secreted by Porphyromonas gingivalis. This evidence of inhibitory activity will encourage the application of M. ciliatum effectively as a functional food or therapeutic agent to prevent periodontal diseases in the early stages.

Porphyromonas gingivalis-derived lipopolysaccharide causes excessive hepatic lipid accumulation via activating NF-κB and JNK signaling pathways.

Porphyromonas gingivalis is the main pathogen of periodontal disease affecting over half of the worldwide adult population. Recent studies have shown that P.gingivalis is related to the development of non-alcoholic fatty liver disease (NAFLD), a global major chronic liver disease, especially in developed countries. However, how P.gingivalis contributes to the pathogenesis of NAFLD has not been fully clarified. We aimed to conduct a preliminary exploration of the underlying mechanism of P.gingivalis infection in the development of NAFLD. Human hepatocellular cells HepG2 were incubated with/without oleic acid (OA) and tested for lipid accumulation upon stimulation by lipopolysaccharide (LPS) derived from P.gingivalis or Escherichia coli. Intracellular lipid droplet formation was analyzed and quantified by Oil Red O staining. The involvement of signaling pathway molecules and pro-inflammatory cytokines related to NF-κB and MAPKs were examined with Western blot and quantitative real-time PCR (qRT-PCR) analyses and further evaluated with inhibitor treatment and RNA interference. HepG2 cells accumulated more intracellular lipids when stimulated with P.gingivalis LPS, as compared to cells treated with E.coli LPS or control. Further pathway analysis demonstrated that after stimulation with P.gingivalis LPS, cells displayed significantly upregulated MyD88 expression, increased phosphorylation of p65 and JNK, and more release of pro-inflammatory cytokines, such as IL-1, IL-8, and TNF-α. In addition, suppression of phosphorylation of p65 and JNK by inhibitors and RNA interference resulted in a reduction in lipid accumulation upon P.gingivalis LPS treatment. These results suggest that P.gingivalis-derived LPS may contribute to intracellular lipid accumulation and inflammatory reaction of HepG2 cells via the activation of NF-κB and JNK signaling pathways. This study offers a possible explanation to the functional involvement of P.gingivalis infection in the pathological progression of NAFLD. These findings may help design new treatment strategies in NAFLD.

Porphyromonas gingivalis induces depression via downregulating p75NTR-mediated BDNF maturation in astrocytes

Many cross-sectional epidemiological studies have shown the incidence of periodontitis is positive correlated with that of depression. However, their causal relationship and underlying mechanism are largely unknown. Porphyromonas gingivalis (Pg) is the main pathogen for periodontitis. Employing female mice treated with Pg every other day for 4 weeks, we found that Pg-mice showed obvious depression-like behavior, an increased number of activated astrocytes and decreased levels of mature brain derived neurotrophic factor (BDNF) and astrocytic p75NTR in the hippocampus. Both hippocampal injection of BDNF and overexpression of p75NTR in astrocytes alleviated Pg-induced depression-like behavior in mice. Moreover, Pg-lipopolysaccharides (LPS) generated similar phenotypes, which were reversed by the TLR-4 inhibitor TAK242. Our results suggest that Pg-LPS decreases the level of astrocytic p75NTR and then downregulates BDNF maturation, leading to depression-like behavior in mice. Our study provides the first evidence that Pg is a modifiable risk factor for depression and uncovers a novel therapeutic target for the treatment of depression.

A natural anti-periodontitis agent, epimedokoreanin B, inhibits virulence activities of gingipains from Porphyromonas gingivalis.

Gingipains are potent virulence cysteine proteases secreted by Porphyromonas gingivalis, a major pathogen of periodontitis. We previously reported that epimedokoreanin B inhibits the activities of gingipains. In this report, we show that epimedokoreanin B inhibits the virulence of gingipains-containing P. gingivalis culture supernatants, indicating the potential use of this prenylated flavonoid as a new agent to combat against periodontal pathogens.

Fretibacterium sp. human oral taxon 360 is a novel biomarker for periodontitis screening in the Japanese population.

BackgroundPeriodontitis is a common inflammatory disease, leading to bone destruction and tooth loss. Screening for periodontitis is important in preventing the progress of this disease. Various types of bacteria have been examined as potential screening targets, but only culturable pathogenic bacteria have been considered candidates. Recently, the various uncultivable bacteria have been identified in microbiome studies, but the value of these bacteria in periodontitis screening remains unknown.ObjectivesThe aim of this study was to evaluate the diagnostic use of uncultivable bacteria Fretibacterium sp. HOT 360 and TM7 sp. HOT 356 for periodontitis screening in the Japanese population.Material and methodsStimulated saliva samples were collected from 217 participants (periodontitis group, n = 157; healthy group, n = 60). The two uncultivable bacterial species selected were: Fretibacterium sp. human oral taxon 360 (Fretibacterium sp. HOT 360) and TM7 sp. human oral taxon 356 (TM7 sp. HOT 356). The levels of these two bacterial species were compared with those of Porphyromonas gingivalis (P. gingivalis), a keystone pathogen in periodontitis. These three species of bacteria were then quantified using qualitative real-time polymerase chain reaction (qPCR) with specific primers and Taqman probes. Statistical analysis was performed by SPSS 20.0 software. P value was statistically significant at .05.ResultsThe populations of uncultivable bacterial species TM7 sp. HOT 356 and Fretibacterium sp. HOT 360 were significantly higher in periodontitis group than in healthy group. Only Fretibacterium sp. HOT 360 showed a significantly positive correlation with such periodontal parameters as probing pocket depth (PPD) and bleeding on probing (BOP).ConclusionThese findings indicate that uncultivable bacteria Fretibacterium sp. HOT 360 can be used as a saliva-based diagnostic bacterial biomarker for periodontitis screening.

Porphyromonas gingivalis Infection Promoted the Proliferation of Oral Squamous Cell Carcinoma Cells through the miR-21/PDCD4/AP-1 Negative Signaling Pathway.

Recent epidemiological studies have revealed that Porphyromonas gingivalis, a major pathogen in periodontal disease, is associated with the development of oral squamous cell carcinoma (OSCC). However, the underlying mechanisms induced by P.gingivalis have not been well-defined. We aimed to determine the role of P.gingivalis in OSCC proliferation and the relevant molecular mechanisms. A cellular proliferation model of OSCC Tca8113 cells infected by P.gingivalis at a multiplicity of infection (MOI) of 50 was established. Cell proliferation was drastically increased in the infected cells compared with the control cells, while the proportion of cells in S phase was increased and the proportion of cells in G1 phase was decreased in the infected cells compared with the control cells. Additionally, the levels of activator protein 1 (AP-1; c-Jun and c-Fos) and its target gene cyclin D1 were increased in P.gingivalis-infected Tca8113 cells compared with control cells. miR-21 expression was elevated when programmed cell death 4 (PDCD4) expression was downregulated. Cyclin D1 expression was regulated by miR-21, PDCD4, and AP-1. The disruption of the pathway by silencing c-Jun, blocking miR-21 expression, or overexpressing PDCD4 led to decreased cyclin D1 expression and inhibited cell proliferation. P.gingivalis DNA levels were positively correlated with miR-21 and c-Jun expression and negatively correlated with PDCD4 expression in clinical OSCC samples. Our findings indicated that P.gingivalis might promote OSCC proliferation by regulating cyclin D1 expression via the miR-21/PDCD4/AP-1 negative feedback signaling pathway.

Research on the roles of genes coding ATP-binding cassette transporters in Porphyromonas gingivalis pathogenicity.

Porphyromonas gingivalis, as a major pathogen of periodontitis, could rapidly adhere to and invade host gingival epithelial cells (GECs) for the induction of infection. One ATP-binding cassette (ABC) transporter gene was found to be upregulated during this infection process, however, the molecular mechanisms remain unclear. In this study, we systemically investigated the messenger RNA level changes of all ABC transporter family genes in P. gingivalis while being internalized within GECs by real-time polymerase chain reaction. We identified that two ABC transporter genes, PG_RS04465 (PG1010) and PG_RS07320 (PG1665), were significantly increased in P. gingivalis after coculturing with GECs. Mutant strains with knockout (KO) of these two genes were generated by homogenous recombination. PG_RS04465 and PG_RS07320 KO mutants showed no change in the growth of bacteria per se. Knockdown of PG_RS07320, but not PG_RS04465, caused decreased endotoxin level in the bacteria. In contrast, both mutant strains showed decreased Arg- and Lys-gingipains activities, with significantly reduced adhesion and invasion capabilities. Secreted interleukin-1β (IL-1β) and IL-6 levels in GECs cocultured with PG_RS04465 or PG_RS07320 KO mutants were also decreased, whereas, only the cells cocultured with PG_RS07320 KO mutants showed significant decrease. In addition, virulence study using mouse revealed that both KO mutant strains infection caused less mouse death than wild-type strains, showing reduced virulence of two KO strains. These results indicated that ABC transporter genes PG_RS04465 and PG_RS07320 are positive regulators of the virulence of P. gingivalis.

Bismuth drugs tackle Porphyromonas gingivalis and attune cytokine response in human cells.

Periodontitis is the leading cause of severe tooth loss and edentulism in adults worldwide and is closely linked to systemic conditions such as diabetes and cardiovascular disease. Porphyromonas gingivalis is the key pathogen in periodontitis. Herein, we provided the first evidence that bismuth drugs suppress P. gingivalis in its planktonic, biofilm, and intracellular states. In total, 42 bismuth-associated proteins were identified including its major virulent factors (e.g., gingipains, hemagglutinin HagA, and fimbriae). Bismuth perturbed its iron acquisition, disturbed the energy metabolism and virulence, and deactivated multiple key enzymes (e.g., superoxide dismutase and thioredoxins). Moreover, bismuth inhibited its biofilm formation and disrupted the 3-day matured biofilms. Notably, the internalized P. gingivalis in various human cells (e.g., human gingival epithelium progenitors, HGEPs) was oppressed by bismuth but not the commonly used antibiotic metronidazole. Importantly, bismuth drugs enabled the counteraction of immuno-inflammatory responses in different host cells perturbed by P. gingivalis. The production of IL-6 and IL-8 attenuated by P. gingivalis in both of native and IL-1β-stimulated HGEPs was restored, while the bacterium-enhanced expression of IL-6, IL-1β, and TNFα in THP-1 macrophages was alleviated. This proof-of-concept study brings prospects for the potential reposition of the routinely used anti-Helicobacter pylori bismuth drugs to better manage inflammatory diseases such as periodontitis and P. gingivalis-related complex systemic disorders.

АЛЬТЕРНАТИВНИЙ ПОГЛЯД ЩОДО ПІДГОТОВКИ ДО ДЕНТАЛЬНОЇ ІМПЛАНТАЦІЇ ПАЦІЄНТІВ МОЛОДОГО ВІКУ ІЗ ХРОНІЧНИМ ЛОКАЛІЗОВАНИМ ПАРОДОНТИТОМ

&#x0D; У статті розглянуті можливості використання бактеріостатиків у передопераційній підготовці пацієнтів молодого віку із хронічним локалізованим пародонтитом без супутньої соматичної патології, що дозволяє ефективно досягти елімінації пародонтальних бактерій та забезпечити умови для повноцінної остеоінтеграції внутрішньокісткових конструкцій.&#x0D;

Aggregatibacter actinomycetemcomitans mediates protection of Porphyromonas gingivalis from Streptococcus sanguinis hydrogen peroxide production in multi-species biofilms

Mixed species biofilms are shaped and influenced by interactions between species. In the oral cavity, dysbiosis of the microbiome leads to diseases such as periodontitis. Porphyromonas gingivalis is a keystone pathogen of periodontitis. In this study, we showed that polymicrobial biofilm formation promoted the tolerance of Porphyromonas gingivalis to oxidative stress under micro-aerobic conditions. The presence of Streptococcus sanguinis, an oral commensal bacterium, inhibited the survival of P. gingivalis in dual-species biofilms via the secretion of hydrogen peroxide (H2O2). Interestingly, this repression could be attenuated by the presence of Aggregatibacter actinomycetemcomitans in tri-species biofilms. It was also shown that the katA gene, encoding a cytoplasmic catalase in A. actinomycetemcomitans, was responsible for the reduction of H2O2 produced by S. sanguinis, which consequently increased the biomass of P. gingivalis in tri-species biofilms. Collectively, these findings reveal that polymicrobial interactions play important roles in shaping bacterial community in biofilm. The existence of catalase producers may support the colonization of pathogens vulnerable to H2O2, in the oral cavity. The catalase may be a potential drug target to aid in the prevention of periodontitis.

Fusobacterium nucleatum stimulates monocyte adhesion to and transmigration through endothelial cells

ObjectiveFusobacterium nucleatum (F. nucleatum) is an important pathogen in periodontitis. Previous studies have demonstrated its ability to spread via haematogenesis and modulate host immune responses. However, little is known regarding its effect on endothelial cells (ECs) and leukocytes. The aim of this study was to assess the effect of F. nucleatum on monocyte attachment and transmigration through ECs. DesignHuman umbilical vein endothelial cells (HUVECs) and human leukemic monocyte (THP-1) cells were infected with F. nucleatum and assessed for monocyte adhesion, transendothelial migration, and HUVEC proliferation/apoptosis. Real-time PCR, western blotting and ELISA were performed to assess the expression of proinflammatory cytokines, adhesion molecules and chemokines in both cells. ResultsF. nucleatum challenge significantly induced THP-1 cell adhesion and transmigration and markedly impaired cell proliferation and apoptosis in HUVECs. A parallel increase in vascular cell adhesion molecule-1, intercellular adhesion molecule-1 and E-selectin expression in HUVECs and an upregulation of tumour necrosis factor-α, interleukin (IL)-1β, IL-6, IL-8, and monocyte chemoattractant protein-1 in both HUVECs and THP-1 cells were observed. The expression of nuclear factor-kappa B was also enhanced in HUVECs and THP-1 cells upon F. nucleatum infection. ConclusionsF. nucleatum triggers an inflammatory response against infection in cells and promotes the recruitment and transmigration of monocytes through ECs.

Castanopsis lamontii Water Extract Shows Potential in Suppressing Pathogens, Lipopolysaccharide-Induced Inflammation and Oxidative Stress-Induced Cell Injury.

Castanopsis lamontii is traditionally used to prevent inflammatory diseases such as periodontitis and pharyngitis by residents in southwest China. However, little scientific evidence has been found to support this. In this research, the antibacterial activities of Castanopsis lamontii water extract (CLE) were assessed using the micro-dilution method. The anti-inflammatory and antioxidant activities of CLE were investigated in RAW264.7 cells. Key bioactive compounds in CLE were also explored. Results showed that CLE was capable of inhibiting the periodontitis pathogen Porphyromonas gingivalis and the pharyngitis pathogen β-hemolytic Streptococcus. It suppressed lipopolysaccharide-induced inflammation in RAW 264.7 cells via inactivating the TLR4/NF-κB pathway. Besides, it reduced oxidative stress-induced cell injury via scavenging reactive oxygen species. Chemical composition analysis revealed that CLE was rich in epicatechin and procyanidin B2. Further studies confirmed that epicatechin predominantly contributed to the antibacterial activities of CLE, while procyanidin B2 was mainly responsible for the anti-inflammatory activities of CLE. Both compounds contributed to the antioxidant activities of CLE. Acute oral toxicity tests proved that CLE was practically non-toxic. These results provide experimental evidences of the health-beneficial effects of CLE and may help promote the application of CLE in the food and health industries.

Hydrogen peroxide masks the bitterness of chlorhexidine mouthwash without affecting its antibacterial activity.

Background:Chlorhexidine (CHX) is an antiseptic mouthwash widely used as the gold standard for inhibiting plaque formation. However, the bitter taste of CHX limits patient compliance. We developed a 0.12% CHX and 1.5% hydrogen peroxide (H2O2) mouthwash that masked the bitter taste of CHX. This study evaluated the antibacterial activity and subject satisfaction of the developed mouthwash.Materials and Methods:Three mouthwashes were used as follows: (1) a commercial 0.12% CHX mouthwash, (2) a prepared 0.12% CHX mouthwash containing 1.5% H2O2, and (3) a prepared 0.12% CHX mouthwash. A disc diffusion assay was performed to determine the antibacterial activity of each mouthwash against Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. To assess subject satisfaction with each mouthwash, a satisfaction questionnaire was completed immediately after rinsing with each mouthwash.Results:The antibacterial activities of the three mouthwashes were similar. Moreover, the questionnaire results revealed that the level of satisfaction was significantly higher for the 0.12% CHX/1.5% H2O2 mouthwash compared with the other mouthwashes.Conclusion:The 0.12% CHX/1.5% H2O2 mouthwash revealed a similar antibacterial activity as the CHX standard against periodontal disease pathogens. In addition, the subjects were more satisfied with the new formula compared with 0.12% CHX alone. These data suggest that the 0.12% CHX/1.5% H2O2 formulation is an alternative antibacterial mouthwash to avoid the unpleasant CHX side effects.

The regulation of Porphyromonas gingivalis biofilm formation by ClpP

Porphyromonas gingivalis is one of the most commonly detected pathogens in periodontal disease and root canal infections. Its viability and pathogenicity are greatly increased in plaque biofilms. Some caseinolytic proteases (Clp) reportedly regulate biofilm formation by various pathogenic bacteria, including P. gingivalis. However, the specific influence of ClpP and its mechanism of regulating biofilm formation by P. gingivalis remains unclear. Hence, in this study, a clpP deletion strain and complemented strain were constructed by homologous recombination, and an in vitro biofilm model was established. Biofilm architecture was observed by scanning electron microscopy. Bacterial cells within the biofilms were examined using confocal scanning laser microscopy. Crystal violet staining was used to determine the amount of formed biofilm. mRNA levels of related regulatory genes were assessed using real-time PCR. The clpP deletion and complemented strains of P. gingivalis were successfully constructed. The biofilm formation ability of the deletion strain was significantly reduced compared with that of the wild-type strain, while that of the complemented strain did not differ from that of the wild-type strain. The expression of fimA, mfa1, and luxS in the deletion strain was lower than in the wild-type and complemented strains at each timepoint. It can be concluded that ClpP increases the biofilm formation of P. gingivalis by regulating the expression levels of fimA, mfa1, and luxS.