Lavandula angustifolia Essential Oil Inhibits the Ability of Fusobacterium nucleatum to Produce Volatile Sulfide Compounds, a Key Components in Oral Malodor.

Oral malodor still constitutes a major challenge worldwide. A strong effort is invested in eliminating volatile sulfur compound-producing oral bacteria through organic natural products such as essential oils. Fusobacterium nucleatum is a known volatile sulfur compound-producing bacteria that inspires oral malodor. The aim of the present study was to test the effect of lavender essential oil on the bacterium's ability to produce volatile sulfide compounds, the principal components of oral malodor. Lavender (Lavandula angustifolia) essential oil was extracted by hydrodistillation and analyzed using GC-MS. The minimal inhibitory concentration (MIC) of lavender essential oil on Fusobacterium nucleatum was determined in a previous trial. Fusobacterium nucleatum was incubated anaerobically in the presence of sub-MIC, MIC, and above MIC concentrations of lavender essential oil, as well as saline and chlorhexidine as negative and positive controls, respectively. Following incubation, volatile sulfur compound levels were measured using GC (Oralchroma), and bacterial cell membrane damage was studied using fluorescence microscopy. Chemical analysis of lavender essential oil yielded five main components, with camphor being the most abundant, accounting for nearly one-third of the total lavender essential oil volume. The MIC (4 µL/mL) of lavender essential oil reduced volatile sulfur compound secretion at a statistically significant level compared to the control (saline). Furthermore, the level of volatile sulfur compound production attributed to 1 MIC of lavender essential oil was in the range of the positive control chlorhexidine with no significant difference. When examining bacterial membrane damage, 2 MIC of lavender essential oil (i.e., 8 µL/mL) demonstrated the same, showing antibacterial membrane damage values comparative to chlorhexidine. Since lavender essential oil was found to be highly effective in hindering volatile sulfur compound production by Fusobacterium nucleatum through the induction of bacterial cell membrane damage, the results suggest that lavender essential oil may be a suitable alternative to conventional chemical-based anti-malodor agents.

Hydroxyethyl Cellulose Promotes the Mucin Retention of Herbal Extracts Active against Streptococcus mutans.

Streptococcus mutans is considered a major cariogenic bacterium. Most anti-cariogenic dentifrices are limited by a short exposure time. The aim of the present study was to test the hypothesis that adding a mucoadhesive agent to the formulation may increase its bioavailability and efficacy. We tested the effect of adding hydroxyethyl cellulose (HEC) to an herbal extract solution containing lavender, echinacea, sage, and mastic gum, which have been previously shown to be effective against Streptococcus mutans. Mucin-coated wells were treated with four test solutions: saline, herbal extracts, herbal extracts with HEC, and chlorhexidine. The wells were incubated with Streptococcus mutans and studied for biofilm formation (Crystal violet assay), acid production (lactate assay), acid tolerance (ATPase assay), and exopolysaccharide (EPS) production using fluorescent microscopy. The results showed that the addition of HEC to the herbal extract solution caused a significant reduction in Streptococcus mutans biofilm formation, lactic acid production, and EPS quantity (p < 0.001). These results suggest that HEC may be a beneficial added excipient to herbal extracts in an anti-cariogenic formulation.

Microbiome Changes in Children Treated under General Anesthesia for Severe Early Childhood Caries: Pilot Study.

A full-mouth radical dental treatment under general anesthesia is a common approach for treating severe early childhood caries (S-ECC). However, previous study showed recurrence of the disease in 80% of cases within 12 months. The aim of the present study was to examine the changes in microbial composition of the dental biofilm of these children following treatment. Dental biofilm samples from five children (mean age 45.4 ± 10.1 months) were taken before and three months after treatment and analyzed for microbial composition using Next Generation Sequencing of the microbial DNA extracted from these samples. Although some reductions in the abundance of caries-pathogenic bacteria (e.g., Streptococcus mutans, Streptococcus sobrinus, Rothia dentocariosa and Scardovia wiggisiae) were seen in the post-treatment follow up samples, these reductions were for the most part not statistically significant, and these bacteria remained well above detection levels. Taken together, the results of the present pilot study suggest that the dental treatment alone is not enough to reduce the caries risk status of these children and that a more comprehensive approach should be considered.

Effect of Denture Base Fabrication Technique on Candida albicans Adhesion In Vitro.

Denture stomatitis is a common manifestation of oral candidiasis affecting some 65% of denture wearers. This condition is initiated by the adherence of Candida albicans to denture base acrylic resin. The present study aimed to test the in vitro effect of traditional and novel fabrication methods on Candida albicans adhesion to denture base samples. Denture based acrylic discs were fabricated using: (i) computerized milling, (ii) 3D printing, (iii) heat curing, and (iv) cold curing. Discs were tested for surface roughness (Ra), hydrophobicity (contact angle), mucin adsorption (Bradford assay), and Candida albicans adhesion. 3D printing significantly increased microbial cell adhesion as compared with heat curing, and computerized milling significantly decreased it. These results were associated with mucin adsorption levels rather than surface roughness. Results suggest that 3D printing may increase the risk for developing denture stomatitis, whereas computerized milling may decrease it as compared with traditional heat curing denture base fabrication.

Effect of Sublethal Blue Light on Herbal Extract Activity Against Volatile Sulfide Compound Production by Fusobacterium nucleatum.

Previously, we have shown that sublethal exposure of blue light caused increased cell membrane permeability in Fusobacterium nucleatum. The aim of the present study was to test the effect of this exposure on the activity of Lavender, Sage, Echinacea and Mastic gum extracts against volatile sulfide compound (VSC) production by Fusobacterium nucleatum. Bacterial suspensions were pre-exposed to blue light (400-500 nm) bellow minimal inhibitory dosage (sub-MID). Exposed and nonexposed samples were inoculated into test tubes containing growth medium, filtered saliva with or without herbal extracts. Following incubation, test tubes were tested for malodor production (odor judge scores), VSC levels (OralChroma), salivary protein degradation (SDS-PAGE) and bacterial cell membrane damage (fluorescence microscopy). Results showed that sub-MID blue light exposure significantly increased the ability of Lavender and Echinacea to reduce VSC production by Fusobacterium nucleatum by more than 30%. These results suggest that sublethal blue light exposure may be useful to increase the efficacy of antimalodor agents.

Effect of High Intensity Blue Light on Fusobacterium nucleatum Membrane Integrity.

Oral malodour is considered to be caused mainly by the production of volatile sulfide compounds (VSC) by anaerobic gram-negative oral bacteria. Previous studies showed that these bacteria were susceptible to blue light phototoxicity mediated by the production of reactive oxygen species (ROS). In the present study, we tested the effect of blue light on the integrity Fusobacterium nucleatum's membrane, cellular proteins and DNA. Bacterial samples were exposed to high intensity blue light for 0, 70, 140 and 280 s (i.e. fluences of 0, 96, 192 and 384 J cm-2 , respectively). Following light exposure, bacterial samples were examined for membrane damage using fluorescence microscopy, intra-cellular protein analysis using electrophoresis (SDS-PAGE) and DNA fragmentation using ultra-filtration. Results showed that the increasing exposure of bacterial samples to blue light caused increased membrane permeability concomitant with a reduction in intra-cellular proteins and DNA fragments content. These results suggest that membrane damage is the main effect of high intensity blue light exposure on malodour producing bacteria.

Mechanism and Prevention of Titanium Particle-Induced Inflammation and Osteolysis.

The worldwide number of dental implants and orthopedic prostheses is steadily increasing. Orthopedic implant loosening, in the absence of infection, is mostly attributable to the generation of wear debris. Dental peri-implantitis is characterized by a multifactorial etiology and is the main cause of implant failure. It consists of a peri-implant inflammatory lesion that often results in loss of supporting bone. Disease management includes cleaning the surrounding flora by hand instruments, ultrasonic tips, lasers, or chemical agents. We recently published a paper indicating that US scaling of titanium (Ti) implants releases particles that provoke an inflammatory response and osteolysis. Here we show that a strong inflammatory response occurs; however, very few of the titanium particles are phagocytosed by the macrophages. We then measured a dramatic Ti particle-induced stimulation of IL1ß, IL6, and TNFα secretion by these macrophages using multiplex immunoassay. The particle-induced expression profile, examined by FACS, also indicated an M1 macrophage polarization. To assess how the secreted cytokines contributed to the paracrine exacerbation of the inflammatory response and to osteoclastogenesis, we treated macrophage/preosteoclast cultures with neutralizing antibodies against IL1ß, IL6, or TNFα. We found that anti-TNFα antibodies attenuated the overall expression of both the inflammatory cytokines and osteoclastogenesis. On the other hand, anti-IL1ß antibodies affected osteoclastogenesis but not the paracrine expression of inflammatory cytokines, whereas anti-IL6 antibodies did the opposite. We then tested these neutralizing antibodies in vivo using our mouse calvarial model of Ti particle-induced osteolysis and microCT analysis. Here, all neutralizing antibodies, administered by intraperitoneal injection, completely abrogated the particle-induced osteolysis. This suggests that blockage of paracrine inflammatory stimulation and osteoclastogenesis are similarly effective in preventing bone resorption induced by Ti particles. Blocking both the inflammation and osteoclastogenesis by anti-TNFα antibodies, incorporated locally into a slow-release membrane, also significantly prevented osteolysis. The osteolytic inflammatory response, fueled by ultrasonic scaling of Ti implants, results from an inflammatory positive feedback loop and osteoclastogenic stimulation. Our findings suggest that blocking IL1ß, IL6, and/or TNFα systemically or locally around titanium implants is a promising therapeutic approach for the clinical management of peri-implant bone loss.

Scaling of titanium implants entrains inflammation-induced osteolysis.

With millions of new dental and orthopedic implants inserted annually, periprosthetic osteolysis becomes a major concern. In dentistry, peri-implantitis management includes cleaning using ultrasonic scaling. We examined whether ultrasonic scaling releases titanium particles and induces inflammation and osteolysis. Titanium discs with machined, sandblasted/acid-etched and sandblasted surfaces were subjected to ultrasonic scaling and we physically and chemically characterized the released particles. These particles induced a severe inflammatory response in macrophages and stimulated osteoclastogenesis. The number of released particles and their chemical composition and nanotopography had a significant effect on the inflammatory response. Sandblasted surfaces released the highest number of particles with the greatest nanoroughness properties. Particles from sandblasted/acid-etched discs induced a milder inflammatory response than those from sandblasted discs but a stronger inflammatory response than those from machined discs. Titanium particles were then embedded in fibrin membranes placed on mouse calvariae for 5 weeks. Using micro-CT, we observed that particles from sandblasted discs induced more osteolysis than those from sandblasted/acid-etched discs. In summary, ultrasonic scaling of titanium implants releases particles in a surface type-dependent manner and may aggravate peri-implantitis. Future studies should assess whether surface roughening affects the extent of released wear particles and aseptic loosening of orthopedic implants.

Long Term Follow-Up of Dental Implants Placed in Autologous Onlay Bone Graft.

PURPOSE: The aim of this study was to evaluate the efficacy of autologous intraoral onlay bone grafting (OBG) in correlation with long-term survival rates of dental implants placed in the augmented bone. MATERIALS AND METHODS: A retrospective study was conducted on 214 patients who received a total of 633 dental implants placed in 224 autologous intraoral block OBG augmentations, combined with Bio-Oss - mixed with platelet-rich plasma (PRP) and covered by platelet-poor plasma (PPP) - as scaffold, with a follow-up time up to 137 months (mean 39.9 ± 30.9 months). RESULTS: A total of 216 OBG cases were successful (96.4%), and most of the augmentations were uneventful (88.4%). Bone graft exposure was moderately associated with bone graft failure (χ(2) = 3.76, p = .052). The healing period after implant placement was 4-6 months (mean 5.6 ± 2.56). The majority of the 591 implants survived (93.4%). The cumulative survival rate of the implants was 83%. CONCLUSIONS: We suggest that augmentation of severely atrophied jaw bone through the placement of horizontal and/or vertical intraoral OBGs in combination with Bio-Oss saturated with PRP and covered by PPP should be considered a reliable, safe, and very effective surgical technique for obtaining high bone graft survival rate and high long-term implant survival rate.

[Halitosis management by the general dental practitioner- results of an International Consensus Workshop*]. / Halitosismanagement für die Zahnarztpraxis. Ergebnisse eines internationalen Konsunsus-Workshops*.

Clinical investigations on patients suffering from halitosis clearly reveal that in the vast majority of cases the source for an offensive breath odor can be found within the oral cavity (90%). Based on these studies, the main sources for intra-oral halitosis where tongue coating, gingivitis/periodontitis and a combination of the two. Thus, it is perfectly logical that general dental practitioners (GDPs) should be able to manage intra-oral halitosis under the conditions found in a normal dental practice. However, GDPs who are interested in diagnosing and treating halitosis are challenged to incorporate scientifically based strategies for use in their clinics. Therefore, the present paper summarizes the results of a consensus workshop of international authorities held with the aim to reach a consensus on general guidelines on how to assess and diagnose patients’ breath odor concerns and general guidelines on regimens for the treatment of halitosis.

Zinc enhances the phototoxic effect of blue light against malodour-producing bacteria in an experimental oral biofilm.

Oral malodour is thought to be caused mainly by the production of volatile sulfide compounds (VSCs) by anaerobic Gram-negative oral bacteria. Previous studies have shown that these bacteria are susceptible to blue light (400-500 nm wavelength). In the present study, we tested the effect of blue light in the presence of zinc, erythrosine B or both on malodour production in an experimental oral biofilm. Biofilms were exposed to a plasma-arc light source for 30, 60 and 120 s (equal to energy fluxes of 41, 82 and 164 J cm(-2), respectively) with or without the addition of zinc acetate, erythrosine B or both. After the light exposure, biofilm samples were examined for malodour production (by an odour judge) and VSC production (with a Halimeter), and VSC-producing bacteria were quantified using a microscopy-based sulfide assay (MSA) and in situ confocal laser scanning microscopy (CLSM). Results showed that exposing experimental oral biofilm to both blue light and zinc reduced malodour production, which coincided with a reduction in VSC-producing bacteria in the biofilm. These results suggest that zinc enhances the phototoxicity of blue light against malodour-producing bacteria.

The effect of a mild increase in temperature on tooth bleaching.

OBJECTIVE: To evaluate the effect of temperature on external tooth bleaching. METHOD AND MATERIALS: Human molars and premolars were internally stained and assessed using a standardized tea solution. The mesial aspects of the teeth were assigned to the treatment group (bleaching at 45 degrees C), and the distal aspects of the teeth were assigned to the control group (bleaching at 37 degrees C). An electronic chromometer with a tristimulus color analyzer was used to measure color change. The mean changes in the values of L*, a*, and b* from baseline to posttreatment were recorded, as were the overall color differences of the specimens in each group (DeltaE*). RESULTS: Increasing the temperature to 45 degrees C significantly (P < .05) increased the bleaching efficacy and reduced the time needed to gain a lighter tooth shade. CONCLUSION: A controlled increase in the temperature of the bleaching agent might be useful in achieving better tooth bleaching in a shorter period of time.

Association between transmucosal depth of osseointegrated implants and malodor production.

PURPOSE: The aim of the present study was to test the association between transmucosal depth of 2-stage dental implants and malodor production. MATERIALS AND METHODS: Fifty-nine 2-stage implants were tested in 14 patients. Measurements were conducted 3 to 4 weeks following second-stage surgery. Measurements included healing abutment malodor scored using a subjective scale, volatile sulfide compounds levels measured using a sulfide monitor (Halimeter), and microbial sampling for anaerobic growth and malodor production. RESULTS: All the malodor-related parameters measured in this study were significantly associated with the transmucosal depth. A significant increase in severity was observed concomitant with the increase in transmucosal depth. CONCLUSION: Based upon the data from this study of 59 two-stage implants in 14 patients, it appears that transmucosal depth of 2-stage dental implants may be an important factor affecting the presence of anaerobic bacterial population and resulting malodor production within the implant-abutment interface.

Effect of liquid-polish coating on in vitro biofilm accumulation on provisional restorations: Part 2.

OBJECTIVE: In part 1 of this study, the authors showed that coating polymethyl methacrylate (PMMA) provisional restorations with bonding resin (BR) or liquid polish (LP) significantly reduced early in vivo biofilm formation on these restorations. The aim of the present study was to determine the mechanism through which these materials inhibit biofilm formation. METHOD AND MATERIALS: The antimicrobial properties of the tested materials were examined using the agar diffusion test (ADT) and the direct contact test (DCT). Surface energy was determined using contact angle measurements; salivary protein adsorption was analyzed by polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: ADT and DCT showed that the tested materials had no antimicrobial properties. Contact angle measurements revealed that liquid polish and PMMA have a similar contact angle, tending toward the hydrophobic region, and that bonding resin was more hydrophilic. SDS-PAGE analysis showed a significant reduction in salivary protein adsorption to the tested materials compared with that to the PMMA control. CONCLUSION: Liquid polish prevents biofilm formation by preventing protein adsorption.

Oral malodor reduction by a palatal mucoadhesive tablet containing herbal formulation.

OBJECTIVE: The aim of the present study was to test the effect of a palatal mucoadhesive tablet containing an herbal formulation on oral malodor production and volatile sulfide compound (VSC) levels, and to evaluate its antimicrobial activity. METHODS: A total of 56 healthy young volunteers participated in experiments 1 and 2. The palatal adhesive tablets were prepared with different active ingredients (herbal formulation, zinc and chlorhexidine), or without an active ingredient as control (placebo). Measurement included odor judge scores (two judges) and VSC readings by a sulfide monitor (Halimeter). In experiment 3, the antimicrobial activity of the herbal formulation ingredients (i.e. sage, Echinacea, Lavender and Mastic gum) were tested against three oral pathogens (Streptoccocus mutans, Porphyromonas gingivalis and Candida albicans) by the agar diffusion test. RESULTS: Application of the palatal adhesive tablets containing herbal formulation resulted in a significant reduction in both oral malodor scores (p<0.001) and VSC levels (p=0.013). Herbal formulation showed higher significance in VSC reduction (p=0.001), as compared to zinc and chlorhexidine (p=0.024 and 0.032, respectively). Sage, Lavender and Mastic gum showed antimicrobial activity against all three oral pathogens. CONCLUSIONS: Results of the present study suggest that the palatal adhesive tablets containing herbal formulation may serve as an effective means of treatment for patients complaining of oral malodor.

Effect of liquid-polish coating on in vivo biofilm accumulation on provisional restorations: part 1.

OBJECTIVE: Biofilm accumulation on provisional restorations may affect the surrounding tissues' integrity. The purpose of this study was to test in vivo biofilm formation on polymethyl methacrylate (PMMA) self-cured acrylic resin provisional crowns. METHOD AND MATERIALS: Three types of PMMA surfaces were tested: (1) polished, (2) polished and coated with bonding agent, and (3) polished and coated with light-cured liquid polish. After 12 hours in the oral cavity, the crown was removed and examined by confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM). RESULTS: Biofilm, 250 Mum thick, was observed with CLSM on the polished acrylic surface. Significantly less bacterial accumulation was observed on the crowns coated with bonding agent, whereas no biofilm was observed on the crowns coated with liquid polish (P <.001). SEM examination confirmed these findings. CONCLUSIONS: Bonding resin or liquid polish coatings significantly reduce early biofilm formation, which in turn might affect the overall plaque accumulation on provisional restorations.

Effect of various natural medicinals on salivary protein putrefaction and malodor production.

OBJECTIVE: Salivary incubation assays are commonly used in oral malodor studies. Using an in vitro model system, the effect of various natural medicinals (i.e., echinacea, propolis, elder, mastic gum, marigold, sage, lavender, thyme, and chamomile) on salivary protein putrefaction and malodor production was examined. METHOD AND MATERIALS: Malodor production levels were scored by an experienced odor judge. Volatile sulfide levels were measured using a sulfide monitor (Halimeter), and salivary protein degradation was determined densitometrically following electrophoresis on polyacrylamide gel (SDS-PAGE). Microbial population was evaluated by viable counts and microscopy. RESULTS: Whereas all of the various medicinals caused some reduction in malodor production from the incubated whole saliva, echinacea and lavender were the most effective. CONCLUSION: The bioassay utilized in the present study suggests that these herbs may inhibit oral malodor production.

Antimicrobial effect of mastic gum methanolic extract against Porphyromonas gingivalis.

The antimicrobial effect of mastic gum, an ancient remedy for oral malodor, against Porphyromonas gingivalis, a known odorogenic periopathogenic oral bacterium, was tested using the agar diffusion test. Paper discs impregnated with mastic gum methanolic extract (MME) [0.5-4% (wt/vol)] produced inhibition zones of 10.5-13.7 mm, respectively, without showing signs of hemolysis, whereas chlorhexidine (0.2%)-impregnated discs, which showed greater inhibition (33.5 mm), also produced large and distinctive hemolytic zones (17 mm). Further analysis of the antimicrobial traits of MME revealed a logarithmic ratio between inhibition zone diameter and MME concentration (r = .99), indicating limited water solubility of this material. These results suggest that mastic gum may be used as a potential nontoxic local agent in treating oral malodor and gum disease.

Effect of visible light on malodour production by mixed oral microflora.

Oral malodour is considered to be caused by the proteolytic activity of anaerobic Gram-negative oral bacteria. In a previous study, it was shown that these bacteria were susceptible to blue light (wavelengths of 400-500 nm). In this study, the effect of blue light on malodour production by mixed oral microflora was tested in a salivary incubation assay. Whole saliva samples were exposed to a xenon light source for 30, 60, 120 and 240 s, equivalent to fluences of 34, 68, 137 and 274 J cm(-2), respectively. Malodour was scored by two judges. The levels of volatile sulfide compounds (VSC) were measured using a sulfide monitor (Halimeter), the microbial population was assessed using viable counts and microscopy, salivary protein degradation was followed by SDS-PAGE densitometry and VSC-producing bacteria were demonstrated using a differential agar. The results showed that the exposure of mixed salivary microflora to blue light caused a reduction in malodour production concomitant with a selective inhibitory effect on the population of Gram-negative oral bacteria. These results suggest that light exposure might have clinical applications for the treatment of oral malodour.