Effectiveness of curcumin-based antimicrobial photodynamic therapy against Staphylococcus aureus.

PURPOSE: This study investigated the effectiveness of curcumin-based antimicrobial photodynamic therapy (aPDT) against Staphylococcus aureus (S. aureus), the causative agent of ventilator-associated pneumonia. METHODS: Curcumin was added to S. aureus culture medium at concentrations of 25, 2.5, and 0.25 µM. After 60 min (20-25°C), each culture was irradiated for 1 and 3 min, and viable bacteria were counted. Curcumin (25 µM) was also added to a bacterial suspension with D-mannitol and sodium azide; microbial counts were determined after irradiation for 3 min. RESULTS: S. aureus was significantly reduced in the 1-min (P = 0.043) and 3-min (P = 0.011) irradiation groups in comparison to the 0-min irradiation group with 25 µM curcumin. No significant differences were observed between the curcumin alone group and the curcumin plus D-mannitol or sodium azide group. CONCLUSION: The findings of this study indicate that prolonged exposure (≥1 min) of S. aureus to LED in 25 µM curcumin solution induces cell wall injury. Curcumin-based aPDT as an adjunct to conventional oral care, employing existing dentistry equipment, offers a promising approach that does not rely on antimicrobial drugs or allows the emergence of resistant bacterial strains. This suggests its potential role in future strategies aimed at preventing ventilator-associated pneumonia.

Impact on Porphyromonas gingivalis of antimicrobial photodynamic therapy with blue light and Rose Bengal in plaque-disclosing solution.

OBJECTIVES: Antimicrobial photodynamic therapy (aPDT) in periodontal pockets using lasers is difficult to perform in some cases because of the high cost of irradiation equipment and the narrow irradiation field. The purpose of the present study was to examine the effects of aPDT in combination with a plaque-disclosing solution and blue light-emitting diode (LED), which are used for composite resin polymerization. METHODS: The reactive oxygen species generated by irradiating 0.001% RB or MB with blue light were analyzed using electron spin resonance spectroscopy. Blue-light exposure was performed at 6.92, 20.76 and 124.6 J. The microorganism to be sterilized was Porphyromonas gingivalis. After aPDT, colony-forming units (CFUs) were measured to estimate cell survival. Carbonylated protein (PC) levels were used to evaluate oxidative stress. All statistical analyses were performed with Tukey's multiple comparisons test or the unpaired t-test. RESULTS: Singlet oxygen (1O2) generation was confirmed by RB+blue LED. 1O2 production was significantly greater with the blue LED irradiation of RB than that of MB (p < 0.0001). CFUs were significantly lower in the blue LED-irradiated group than in the non-LED-irradiated group (p < 0.01). The bactericidal effect increased in a time-dependent manner. aPDT increased PC levels. No morphological changes were observed in P. gingivalis. CONCLUSIONS: The present results suggest that aPDT exerts bactericidal effects against P. gingivalis by increasing oxidative stress through the generation of 1O2 in cells. Periodontal disease may be treated by aPDT using the equipment available in dental offices.

Inhibitory effect of omega-3 fatty acids on alveolar bone resorption and osteoclast differentiation.

In this study, a Porphyromonas gingivalis (P.g.)-infected mouse periodontitis model was used to investigate the effect of omega-3 fatty acid intake on differentiation and maturation of cultured osteoclast. Four-week-old C57BL/6JJcl mice were divided into four groups according to the diets they were fed from the beginning of the experiment (i.e., food containing omega-3 or omega-6 fatty acids) and whether they were orally administered P.g. Thirty-three days after beginning the experiment, bone marrow cells were sampled from the femoral bone of mice from each group and differentiated into osteoclasts; the effects of the ingestion of different fatty acids were subsequently investigated. There was no statistical interaction between the different fatty acids and P.g. infection on the number of osteoclasts (P = 0.6). However, the fatty acid type affected the number of osteoclasts in mice (P = 0.0013), with the omega-3 groups demonstrating lower osteoclast numbers than the omega-6 groups. Furthermore, the addition of resolvin E1 (RvE1), which is an omega-3 fatty acid-derived lipid mediator, suppressed the differentiation of mouse cultured osteoclasts (P < 0.0001). Therefore, the ingestion of omega-3 fatty acids may suppress osteoclast differentiation while inhibiting bone resorption and tissue destruction due to periodontitis.

Kampo Therapies and the Use of Herbal Medicines in the Dentistry in Japan.

Dental caries and periodontal disease are two major diseases in the dentistry. As the society is aging, their pathological meaning has been changing. An increasing number of patients are displaying symptoms of systemic disease and so we need to pay more attention to immunologic aggression in our medical treatment. For this reason, we focused on natural products. Kampo consists of natural herbs-roots and barks-and has more than 3000 years of history. It was originated in China as traditional medicine and introduced to Japan. Over the years, Kampo medicine in Japan has been formulated in a way to suit Japan's natural features and ethnic characteristics. Based on this traditional Japanese Kampo medicine, we have manufactured a Kampo gargle and Mastic Gel dentifrice. In order to practically utilize the effectiveness of mastic, we have developed a dentifrice (product name: IMPLA CARE) and treated implant periodontitis and severe periodontitis.

Effects of French Pine Bark Extract Chewing Gum on Oral Malodor and Salivary Bacteria.

Frequent or persistent malodor (halitosis) represents a considerable embarrassment to those affected. French pine bark extract, Pycnogenol® (PYC), has displayed antibacterial activity against a broad range of bacterial species. In the present study, anticipated benefits of PYC on diminishing halitosis were investigated. Ten healthy males and 11 females, aged 40.1±12.3 y, were recruited based on threshold breath sulfur compounds presence, diagnosed by portable gas chromatography. Subjects were randomly assigned to either sugar-free gums, or gums bearing an additional 2.5 mg PYC per piece. The subjects were required to consume two pieces of PYC or placebo gum six times daily for 15 min. The levels of volatile sulfur compounds (VSCs), measured by OralChromaTM, and tongue-coating score were recorded at baseline, 2, and 4 wk. Hydrogen sulfide-producing bacteria in saliva were cultured on Brucella blood agar plates containing 0.05% cysteine, 0.12% glutathione, and 0.02% lead acetate. The group consuming PYC chewing gum reduced exhaled hydrogen sulfide, methyl mercaptan and dimethyl sulfide significantly (p<0.01) after 2 wk versus baseline. Continuation of daily PYC-gum consumption for 4 wk remarkably lowered the tongue-coating score and exhaled hydrogen sulfide was significantly decreased compared to the placebo group. PYC chewing gum significantly reduced hydrogen sulfide-producing bacteria in saliva after 4 wk (p<0.01), with no effects observed in the placebo control. The results suggest that PYC chewing gum is effective in reducing oral malodor by decreasing the accumulation of tongue coating and the number of hydrogen sulfide-producing bacteria in saliva.

Antibacterial Action of a Condensed Tannin Extracted from Astringent Persimmon as a Component of Food Addictive Pancil PS-M on Oral Polymicrobial Biofilms.

The purpose of this study was to evaluate the antibacterial activity against polymicrobial (PM) biofilms of a condensed tannin extracted from astringent persimmon (PS-M), which is contained in refreshing beverages commercially available in Japan. Salivary PM biofilms were formed anaerobically on glass coverslips for 24 and 72 h and were treated for 5 min with sterilized deionized water (DW), 0.05 and 0.2 wt% chlorhexidine digluconate (CHX), and 0.5-4.0 wt% PS-M solution. The colony forming units (CFU/mL) were determined and morphological changes of the biofilms were observed by scanning electron microscopy (SEM). The CFUs were lower in all PS-M and CHX groups compared to the DW group. PS-M exerted a dose-dependent effect. PS-M (1.53 × 10(7)) at a dose of 4.0 wt% had the same effect as 0.2 wt% CHX (2.03 × 10(7)), regardless of the culture period. SEM revealed the biofilm structures were considerably destroyed in the 4.0 wt% PS-M and 0.2 wt% CHX. These findings indicate that the antibacterial effects of PS-M, a naturally derived substance, are comparable to those of CHX. PS-M may keep the oral cavity clean and prevent dental caries and periodontal disease related to dental plaque, as well as systemic disease such as aspiration pneumonitis.

Inhibitory effects of French pine bark extract, Pycnogenol®, on alveolar bone resorption and on the osteoclast differentiation.

Pycnogenol(®) (PYC) is a standardized bark extract from French maritime pine (Pinus pinaster Aiton). We examined the inhibitory effects of PYC on alveolar bone resorption, which is a characteristic feature of periodontitis, induced by Porphyromonas gingivalis (P. gingivalis) and osteoclast differentiation. In rat periodontitis model, rats were divided into four groups: group A served as the non-infected control, group B was infected orally with P. gingivalis ATCC 33277, group C was administered PYC in the diet (0.025%: w/w), and group D was infected with P. gingivalis and administered PYC. Administration of PYC along with P. gingivalis infection significantly reduced alveolar bone resorption. Treatment of P. gingivalis with 1 µg/ml PYC reduced the number of viable bacterial cells. Addition of PYC to epithelial cells inhibited adhesion and invasion by P. gingivalis. The effect of PYC on osteoclast formation was confirmed by tartrate-resistant acid phosphatase staining. PYC treatment significantly inhibited osteoclast formation. Addition of PYC (1-100 µg/ml) to purified osteoclasts culture induced cell apoptosis. These results suggest that PYC may prevent alveolar bone resorption through its antibacterial activity against P. gingivalis and by suppressing osteoclastogenesis. Therefore, PYC may be useful as a therapeutic and preventative agent for bone diseases such as periodontitis.

Ameliorating effects of Juzentaihoto on restraint stress and P. gingivalis-induced alveolar bone loss.

OBJECTIVE: Juzentaihoto (JTX) is a traditional Japanese medicine that consists of 10 herbs. The purpose of this study was to evaluate the efficacy of multi-herbal medicine JTX as a preventive and therapeutic drug for periodontal bone resorption and for reducing restraint stress. MATERIALS AND METHODS: Porphyromonas gingivalis ATCC 33277 was used for testing the antibacterial activity of JTX and a rat experimental periodontitis model. To evaluate the effect of JTX against P. gingivalis infection, we determined the differences in alveolar bone loss among experimental groups. The concentrations of adrenocorticotropic hormones were measured as stress markers, and atrophy of the thymus and spleen was assessed. RESULTS: JTX had antibacterial activity against P. gingivalis ATCC 33277. JTX treatment of mouse bone marrow cells at a concentration of 0.1 µg/ml significantly inhibited osteoclast formation. Administration of JTX to rats with P. gingivalis infection and restraint stress significantly reduced alveolar bone loss compared with the case with just the combination of P. gingivalis infection and restraint stress. In the restrained groups, stress markers were elevated, and the thymus and spleen were atrophied. The groups with administration of JTX showed not only inhibition of the decrease of weight but also normalization of corticosterone and cortisol values. CONCLUSION: JTX effectively inhibited restraint stress and osteoclastogenesis. It appears that the effects of JTX inhibit the destruction of periodontal tissue by suppressing stress. Our study demonstrated that JTX affects the correlation between restraint stress and periodontitis.

Reactive oxygen species scavenging activity of Jixueteng evaluated by electron spin resonance (ESR) and photon emission.

Jixueteng, the dried stem of Spatholobus suberectus Dunn (Leguminosae), is a traditional Chinese herbal medicine that is commonly classified as a herb that promotes blood circulation and can be used to treat blood stasis. The aim of this study was to examine the reactive oxygen species (ROS) scavenging activity of Jixueteng and other herbal medicines. The ROS scavenging activities of the water extracts of Jixueteng, Cnidium officinale and Salvia miltiorrhiza were examined using an electron spin resonance (ESR) technique and faint luminescence measurement. The ESR signal intensities of the superoxide anion (O2·) and hydroxyl radical (HO·) were reduced more by Jixueteng than the other herbal medicines we tested. High photon emission intensity to hydrogen peroxide (H202) and HO· was observed in Jixueteng using the XYZ chemiluminescence system that was used as faint luminescence measurement and analysis. The results of the present study revealed that the ROS scavenging activity of 8% Jixueteng was the strongest among the herbal medicines we tested. It has been reported that Jixueteng includes various polyphenols. In the ROS scavenging activity by Jixueteng, it is supposed that the antioxidant activity caused by these polyphenols would contribute greatly. In conclusion, a water extract component of Jixueteng had potent free radical scavenging activity and an antioxidative effect that inhibited the oxidative actions of O2·â», H2O2 and HO·. Therefore, Jixueteng represents a promising therapeutic drug for reactive oxygen-associated pathologies.

Inhibitory effects of Jixueteng on P. gingivalis-induced bone loss and osteoclast differentiation.

OBJECTIVE: The aim of this study was to investigate the possibility of Jixueteng as a preventive and therapeutic drug for the periodontitis. We investigated the inhibitory effects of Jixueteng on Porphyromonas gingivalis-induced bone loss in mice, antibacterial activity against P. gingivalis and differentiation of osteoclast and viability of cells. MATERIALS AND METHODS: Fifty-four male, 4-week-old C57BL/6N mice, were randomly divided into the following three groups of 18 mice each; group A served as the P. gingivalis non-infected control (sham group), group B was infected orally with P. gingivalis and group C was administered Jixueteng extract in drinking water and was then infected with P. gingivalis. In order to evaluate the effect of Jixueteng, the distance from the alveolar bone crest to the cemento-enamel junction was determined. P. gingivalis suspension was exposed for 1, 15 and 60 min to 5 ml of the Jixueteng extract. Furthermore, to clarify the mechanism of the inhibitory effects of Jixueteng on osteoclast formation, Jixueteng extract was added to the culture of mouse bone marrow cells, osteoclast precursor. RESULTS: Administration of Jixueteng along with P. gingivalis infection significantly reduced alveolar bone loss compared to P. gingivalis infection. Jixueteng treatment at the concentration of 0.01% significantly inhibited osteoclast formation. The addition of Jixueteng extract (0.1%, 0.01%, and 0.001%) to the culture showed a significant inhibition of the number of surviving osteoclasts in a dose-dependent manner. CONCLUSION: Jixueteng has an antibacterial activity against P. gingivalis and inhibitory effects on osteoclastogenesis, it may be useful as a therapeutic drug in the treatment of P. gingivalis-induced periodontitis.

Quest for anti-inflammatory substances using IL-1ß-stimulated gingival fibroblasts.

BACKGROUND: We have previously reported that azulene-related compounds, and alkaline extract of Sasa senanensis Rehder potently inhibited nitric oxide (NO) production by lipopolysaccharide (LPS)-stimulated mouse macrophages. We investigated here whether they can inhibit pro-inflammatory cytokine production, by activated human gingival fibroblast (HGF). MATERIALS AND METHODS: HGF was established from the periodontal tissues of extracted tooth. Viable cell number was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Production of Prostaglandin E(2) (PGE(2)) and cytokines was determined by enzyme immunoassay, and enzyme-linked immunosorbent assay, respectively. RESULTS: Interleukin (IL)-1ß did not inhibit, but rather slightly stimulated the growth of HGF cells. IL-1ß stimulated the production of PGE(2), IL-6, IL-8 and monocyte chemotactic protein-1 very potently, but not that of nitric oxide and tumor necrosis factor-α. Native LPS and synthetic lipid A from E. coli and P. gingivalis was much less stimulatory. Dexamethasone, not indomethacin, was an efficient inhibitor of IL-8 production. Among five azulene-related compounds, benzo[b]cyclohepta[e][1,4]thiazine most potently inhibited the IL-8 production by HGF cells, as well as NO production by activated RAW264.7 cells. The alkaline extract of Sasa senanensis Rehder significantly inhibited IL-8 production, without affecting the cell viability. CONCLUSION: The present system may be applicable for use in the search for anti-gingivitis substances.