Quantitative PCR Effectively Quantifies Triazole-Susceptible and Triazole-Resistant Aspergillus fumigatus in Mixed Infections.

Increasing resistance to triazole antifungals in Aspergillus fumigatus is worrisome because of the associated high mortality of triazole-resistant A. fumigatus (TRAF) infections. While most studies have focused on single triazole-susceptible (WT) or TRAF infections, reports of TRAF cases developing mixed WT and TRAF infections have been described in several studies. However, the prevalence of mixed infections and their responses to current recommended therapies are unknown and could be inappropriate, leading to poor clinical outcomes. To address the urgent need for tools to diagnose, monitor disease development and therapy efficacies in mixed infection settings where quantification of WT versus TRAF is key, this study developed a novel qPCR assay to differentiate WT and TRAF harboring the cyp51A-TR34/L98H mutation. The proposed assay successfully quantified A. fumigatus and discriminated TRAF-TR34 in vitro and in vivo, which was achieved by increasing the yield of extracted DNA through improved homogenization and specific primers targeting the WT-sequence or TR34-insertion and a TaqMan-probe directed to A. fumigatus. The here-developed qPCR assay overcomes sensitivity issues of methodologies such as CFU counts, providing specific, reproducible, and reliable quantitative information to study and follow up the (interplay and individual) effects of mixed A. fumigatus infections on disease development and treatment responses.

Antimicrobial Mechanisms of Leucocyte- and Platelet Rich Fibrin Exudate Against Planktonic Porphyromonas gingivalis and Within Multi-Species Biofilm: A Pilot Study.

Leucocyte- and platelet rich fibrin (L-PRF) is an autologous biomaterial used in regenerative procedures. It has an antimicrobial activity against P. gingivalis although the mechanism is not fully understood. It was hypothesized that L-PRF exudate releases hydrogen peroxide and antimicrobial peptides that inhibit P. gingivalis growth. Agar plate and planktonic culture experiments showed that the antimicrobial effect of L-PRF exudate against P. gingivalis was supressed by peroxidase or pepsin exposure. In developing multi-species biofilms, the antimicrobial effect of L-PRF exudate was blocked only by peroxidase, increasing P. gingivalis growth with 1.3 log genome equivalents. However, no effect was shown on other bacteria. Pre-formed multi-species biofilm trials showed no antimicrobial effect of L-PRF exudate against P. gingivalis or other species. Our findings showed that L-PRF exudate may release peroxide and peptides, which may be responsible for its antimicrobial effect against P. gingivalis. In addition, L-PRF exudate had an antimicrobial effect against P. gingivalis in an in vitro developing multi-species biofilm.

A dual-strain Lactobacilli reuteri probiotic improves the treatment of residual pockets: A randomized controlled clinical trial.

AIM: To examine the adjunctive effect of a Lactobacillus reuteri probiotic (ATCC PTA 5289 & DSM 17938) on the re-instrumentation of residual pockets. MATERIALS AND METHODS: This randomized, double-blind, placebo-controlled study included 39 previously non-surgically treated periodontitis patients. A re-instrumentation was carried out, and probiotic and/or placebo drops were applied according to the study protocoll. Patients afterwards received lozenges to use 2×/day for 12 weeks. Probing pocket depth (PPD), recession, bleeding on probing and plaque levels were analysed, next to the microbiological impact. RESULTS: No effects of the probiotic drops could be found. However, after 24 weeks, the overall PPD in the probiotic lozenges group (2.64 ± 0.33 mm) was significantly lower compared to the control lozenges (2.92 ± 0.42 mm). This difference was even more pronounced in moderate (4-6 mm) and deep (≥7 mm) pockets. In the probiotic lozenges group, there were also significantly more pockets converting from ≥4 mm at baseline to ≤3 mm at 24 weeks (67 ± 18% versus 54 ± 17%) and less sites in need for surgery (4 ± 4% versus 8 ± 6%). However, the probiotic products did not influence the microbiological counts of the periodontopathogens. CONCLUSION: The adjunctive consumption of L. reuteri lozenges after re-instrumentation improved the PPD reduction, without an impact on pocket colonization with periodontopathogens.

The usage of a lactobacilli probiotic in the non-surgical therapy of peri-implantitis: A randomized pilot study.

OBJECTIVES: Examine the clinical and microbiological benefits of a dual-strain Lactobacillus reuteri probiotic on the non-surgical therapy of initial peri-implantitis. MATERIALS AND METHODS: This randomized, double-blind study targeted patients with initial peri-implantitis, that is peri-implantitis with a maximum mean probing pocket depth of 6 mm and maximum 3 mm bone loss compared with loading. A full-mouth prophylaxis was performed and the peri-implantitis sites were debrided. Subsequently, local application of the study drops was carried out at the peri-implantitis sites and the study lozenges were handed out. The patients in the probiotic group received drops and lozenges containing L. reuteri (ATCC PTA 5289 & DSM 17938), those in the control group received placebo products. At the implant level the measurements of interest were bleeding, probing pocket depth and plaque. Full-mouth bleeding and plaque scores were also recorded. Microbiological samples were taken from the tongue, saliva and subgingivally around the implants. RESULTS: All clinical parameters were significantly decreased after 12 and 24 weeks. At the implant level the only statistically significant difference was a greater decrease in plaque levels in the probiotic versus the control group (p = .002 at 24 weeks). At the full-mouth level, the only intergroup difference was the greater decrease in full-mouth bleeding on probing sites in the probiotic group compared with the control group (p < .001 at 24 weeks). Concerning the microbiological outcomes, no significant differences could be found at any time point, neither intra- nor intergroup. CONCLUSIONS: No adjunctive effects of the use of L. reuteri probiotics in the treatment of peri-implantitis were found.

Terpinen-4-ol and carvacrol affect multi-species biofilm composition.

The aim of this study was to investigate the cytotoxic activity and inhibitory effect of terpinen-4-ol (T4ol) and carvacrol against single- and multi-species biofilms. The toxicity of each compound was tested on oral keratinocytes and evaluated by XTT assay. Inhibition and eradication of single-species biofilms were analyzed by crystal violet assay and the effect on multi-species biofilm composition was evaluated by qPCR. T4ol and carvacrol did not affect the epithelial cell viability, in contrast to chlorhexidine, which showed a high cytotoxic effect. Inhibition and eradication of single-species biofilms treated with T4ol and carvacrol were observed. The same inhibitory effect was observed for multi-species biofilms, especially on periodontal pathogens. In conclusion, specific concentrations of T4ol and carvacrol without toxicity towards the epithelial cells reduced the numbers of periodontal pathogens in single- and multi-species biofilms.

Necrotrophic growth of periodontopathogens is a novel virulence factor in oral biofilms.

The oral use of antimicrobial agents embedded in toothpastes and mouth rinses results in an oral microbial massacre with high amounts of dead bacteria in close proximity to few surviving bacteria. It was hypothesized that this provides the surviving pathogenic bacteria a large amount of dead microbial biomass as a nutritional source for growth (necrotrophy). This study demonstrated the necrotrophic growth of periodontal pathogens in the presence of different dead oral species. In addition, the presence of dead bacteria resulted in an outgrowth of several periodontal pathogens in complex multi-species biofilms. Additionally, upon contact with dead oral bacteria, virulence genes of P. intermedia and P. gingivalis were up-regulated (necrovirulence). This resulted in a more pronounced epithelial cytotoxicity (necrotoxicity). These findings indicate that presence of dead bacteria induce necrotrophy, necrovirulence and necrotoxicity in several oral pathogens.

The effect of a streptococci containing probiotic in periodontal therapy: a randomized controlled trial.

AIM: To evaluate the adjunctive effects of a Streptococcus oralis KJ3, Streptococcus uberis KJ2 and Streptococcus rattus JH145 containing probiotic tablet after scaling and root planing (SRP). MATERIALS AND METHODS: Forty-eight periodontitis patients were included in this double-blind, placebo-controlled clinical trial. After root planing, patients used either a placebo or a probiotic tablet twice a day for 12 weeks. The pocket probing depth (primary outcome measure), bleeding on probing and relative attachment levels were measured at baseline, 12 and 24 weeks. At baseline, 4, 8, 12 and 24 weeks, microbiological sampling was performed and plaque and gingival indices were recorded. RESULTS: The primary and secondary outcome measures were significantly (p < 0.05) improved at the 12- and the 24-week evaluation in both groups. However, no significant inter-group differences could be detected at any time point, except from the % of sites with plaque that were significantly lower in the probiotic group than in the control group at the 24-week evaluation. In addition, at the 12-week time point, the salivary Prevotella intermedia counts were significantly lower in the probiotic group. CONCLUSIONS: No differences were detected when comparing the adjunctive use of a placebo or the investigated streptococci containing probiotic tablet after SRP. ClinicalTrials.gov Identifier: NCT02403960.

Effect of Bdellovibrio bacteriovorus HD100 on multispecies oral communities.

The predation of Bdellovibrio bacteriovorus on different periodontal pathogens has already been described. However, it is necessary to consider the polymicrobial nature of periodontal disease. The current study explores the predation of Bdellovibrio on oral pathogens organized in multispecies communities. The effect of the predator was evaluated on in vitro six species communities with microbial culturing. Additionally, the effect on ex vivo subgingival plaque and saliva samples from periodontitis patients was assessed. In the latter experiment results were examined with microbial culturing, quantitative polymerase chain reaction (qPCR) and denaturing gradient gel electrophoresis (DGGE). The latter technique was used to get an overview of the whole mixed microbial population. Results showed that even in more complex models, B. bacteriovorus was still able to predate on Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. However predation on Prevotella intermedia and Porphyromonas gingivalis could not be validated in multispecies models. The effect of Bdellovibrio was not restricted to the target bacteria. Changes in the overall ecology of the different models were evident. It could be concluded that the efficiency of predation decreased when complexity of the models increased. However, B. bacteriovorus was able to attack two important oral pathogens, F. nucleatum, and A. actinomycetemcomitans, even when present in ex vivo clinical samples. These effects still have to be validated in in vivo models to see the impact of Bdellovibrio on the whole bacterial ecology.

Clinical and microbiological effects of Lactobacillus reuteri probiotics in the treatment of chronic periodontitis: a randomized placebo-controlled study.

AIM: The aim of this randomized placebo-controlled clinical trial was to evaluate the effects of Lactobacillus reuteri-containing probiotic lozenges as an adjunct to scaling and root planing (SRP). MATERIAL AND METHODS: Thirty chronic periodontitis patients were recruited and monitored clinically and microbiologically at baseline, 3, 6, 9 and 12 weeks after therapy. All patients received one-stage full-mouth disinfection and randomly assigned over a test (SRP + probiotic, n = 15) or control (SRP + placebo, n = 15) group. The lozenges were used two times a day for 12 weeks. RESULTS: At week 12, all clinical parameters were significantly reduced in both groups, while there was significantly more pocket depth reduction (p < 0.05) and attachment gain (p < 0.05) in moderate and deep pockets; more Porphyromonas gingivalis reduction was observed in the SRP + probiotic group. CONCLUSIONS: The results indicate that oral administration of L. reuteri lozenges could be a useful adjunct to SRP in chronic periodontitis.

Bacterial antagonism against periodontopathogens.

BACKGROUND: The aim of the current study is to compare the prevalence of commensal bacteria, with beneficial properties, for healthy and diseased individuals and additionally to examine the inhibitory effect of some commercial dietary probiotics on periodontopathogens, comparing this inhibitory effect to that of orally derived beneficial bacteria. METHODS: Subgingival plaque samples from 35 patients (healthy and periodontitis patients) were analyzed. Growth inhibition of the periodontal pathogens Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans was examined using the agar overlay technique and agar well diffusion method. The quantification of the inhibitory effect was checked with the agar well diffusion method. RESULTS: Using the agar overlay technique, the prevalence of strains antagonistic toward P. gingivalis, A. actinomycetemcomitans, and F. nucleatum was found to be higher in healthy individuals than in individuals with periodontitis, but this could not be validated by the agar well diffusion assay. Compared with the antagonistic activity of the isolated strains, the probiotic strains overall showed a stronger inhibition of the periodontal pathogens. CONCLUSION: It was shown that some oral bacteria can cause antagonism toward periodontopathogens, and these observations underline the therapeutic potential of applications that stimulate oral health by the application of beneficial effector strains.

Microbiological outcome of two screw-shaped titanium implant systems placed following a split-mouth randomised protocol, at the 12th year of follow-up after loading.

PURPOSE: To compare the subgingival microbiota around two differently designed implant systems that were in function for more than 12 years in a randomised split-mouth study design, and to compare the outcome with natural dentition. MATERIALS AND METHODS: A total of 18 partially edentulous patients received at least two TiOblast™ (Astra Tech) and two Brånemark (Nobel Biocare) implants following a split-mouth design. At the last follow-up visit, periodontal parameters (probing depth, bleeding on probing and plaque) were recorded and intraoral radiographs were taken to calculate bone loss. Subgingival plaque samples were collected for culture, qPCR and checkerboard DNA-DNA hybridisation analysis. These data were related to implant design and bone loss. This study setup allowed a comparison of 34 Astra Tech (Impl A) with 32 Brånemark (Impl B) implants. RESULTS: During the 12-year follow up, five patients dropped out. One Brånemark implant was lost before abutment connection in a dropout patient. Mean bone loss between loading and year 12 was 0.7 mm (range: -0.8-5.8) (Impl A), and 0.4 mm (range: -1.1-4.1) (Impl B). No significant microbiological differences (qualitative and quantitative) could be observed between both implant types. Compared to teeth, subgingival plaque samples from implants did not reach the concentration of pathogens, even after 12 years of function. CONCLUSIONS: These data show that both implant systems (with differences in macro-design and surface characteristics), in patients with good oral hygiene and a stable periodontal condition, can maintain a successful treatment outcome without significant subgingival microbiological differences after 12 years of loading. The presence of periodontopathogens did not necessarily result in bone loss.

Do periodontopathogens disappear after full-mouth tooth extraction?

AIM: To monitor the intra-oral microbiological changes after full-mouth extraction using quantitative polymerase chain reaction (qPCR). MATERIAL AND METHODS: Nine patients with severe, aggressive periodontitis, for whom a full-mouth tooth extraction was the only remaining treatment option were recruited. Before and 6 months after extraction, microbial samples were obtained (tongue, saliva and subgingival plaque) and analysed by qPCR. RESULTS: The elimination of subgingival niches, by extraction of all natural teeth, resulted in a 3-log reduction of Porphyromonas gingivalis and Tannerella forsythia, and more modest reductions of Aggregatibacter actinomycetemcomitans and Prevotella intermedia. However, the detection frequencies of these periodontopathogens in saliva and on the tongue remained unchanged after full-mouth tooth extraction. CONCLUSION: In contrast to what has been believed so far, full-mouth tooth extraction does not result in eradication of all periodontopathogens but only in a significant reduction. The clinical consequences of this observation remain speculative.

Microbial adhesion on different bracket types in vitro.

OBJECTIVE: To test the hypothesis that there are differences in total bacterial counts and capacity for biofilm formation between seven different bracket types. MATERIAL AND METHODS: By means of an in vitro experiment, seven commercially available bracket systems (Damon [A], Clarity [B], Mystique [C], Speed [D], Victory MBT [E], Micro-loc [F], and Generus [G]) were compared. A total of 25 premolar brackets of each bracket system were incubated in brain heart infusion medium containing the saliva and bacteria of two orthodontic patients. After 72 hours, the amounts of aerobe and anaerobe bacteria were determined by counting the colony-forming units (CFU). The CFU ratio (aerobe/anaerobe) also was calculated, and the black pigmented bacteria were analyzed. RESULTS: Significant differences between the different bracket types in terms of biofilm formation were found. Bracket types can be arbitrarily divided into low, intermediate, and high plaque-retaining brackets. The group with low adhesion consists of bracket types E, F, and G; the group with high adhesion of bracket types A, B, and C; and type D exhibits intermediate adhesion. The group with high microbial adhesion (A, B, and C) did present significantly lower CFU ratios (aerobe/anaerobe) than were exhibited by the other bracket systems (P < .05). CONCLUSION: The hypothesis is accepted. Orthodontic brackets serve as different loci for biofilm formation; in this in vitro study, significant differences were noted between the different types of brackets.

Benefit of "one-stage full-mouth disinfection" is explained by disinfection and root planing within 24 hours: a randomized controlled trial.

OBJECTIVES: The beneficial effects of the one-stage, full-mouth disinfection remain controversial in the scientific literature. This might be due to the fact that an entire mouth disinfection with the use of antiseptics has been confused with a full-mouth scaling and root planing. This parallel, single blind RCT study aimed to compare several full-mouth treatment strategies with each other. MATERIAL AND METHODS: Seventy-one patients with moderate periodontitis were randomly allocated to one of the following treatment strategies: scaling and root planing, quadrant by quadrant, at two-week intervals (negative control, NC), full-mouth scaling and root planing within 2 consecutive days (FRP), or three one-stage, full-mouth disinfection (FM) protocols within 2 consecutive days applying antiseptics to all intra-oral niches for periopathogens using as antiseptics: chlorhexidine (FMCHX) for 2 months, amine fluoride/stannous fluoride for 2 months (FMF), or chlorhexidine for 2 months followed by amine fluoride/stannous fluoride for another 6 months (FMCHX+F). At baseline and after 2, 4, and 8 a series of periodontal parameters were recorded. RESULTS: All treatment strategies resulted in significant (p<0.05) improvements of all clinical parameters over the entire duration of the study. Inter-treatment differences were often encountered. The NC group nearly always showed significant smaller improvements than the two CHX groups. The differences between the FRP or FM groups, and the two CHX groups only sporadically reached a statistical significance. CONCLUSION: These observations indicate that the benefits of the "OSFMD" protocol are partially due to the use of the antiseptics and partially to the completion of the therapy in a short time.

Microbiological and clinical outcomes and patient satisfaction for two treatment options in the edentulous lower jaw after 10 years of function.

BACKGROUND: Long-term data on microbiological and clinical outcome as well as on patient satisfaction after implant therapy in the edentulous mandible are limited. Especially comparisons between fixed full prostheses (FFPs) and overdentures (ODs), or between anchoring systems for the latter are scarce. AIM: This study aimed to evaluate both of these parameters at the 10-year follow-up in a group of fully edentulous patients rehabilitated via an OD or a FFP (the latter to allow inter-group comparison). MATERIAL AND METHODS: A total of 37 fully edentulous patients (25 ODs, 12 FFPs, age at implant installation ranged from 36 to 85 years) participated in this study. All subjects received their implants (Branemark System, Nobel Biocare AB, Gothenburg, Sweden) 10 years previously. For the ODs different attachment systems (bar, magnets, ball) had been applied that allowed a further intra-group comparison. At the follow-up visit, 10 years after the abutment insertion, a series of periodontal parameters were recorded, long-cone radiographs were taken and subgingival plaque samples were collected for analysis using checkerboard DNA-DNA hybridization. The clinical and radiographic data were recorded at abutment connection and after 1 and 10 years. RESULTS: After 10 years of loading, mean plaque and bleeding indices and changes in attachment or marginal bone level were not significantly different, neither between the OD and FFP group, nor within the OD group. The marginal bone loss between abutment connection and year 10 was 0.86 and 0.73 mm for OD and FFP groups, respectively. The subgingival microbiota at implant sites from all (sub)-groups was comparable, with low numbers of DNA counts (+/-10 x 10(5)) but high detection frequencies of Actinobacillus actinomycetemcomitans (>90%), Porphyromonas gingivalis (>85%) and Tannerella forsythensis (30%). The composition of the subgingival microbiota was influenced by probing depth and bleeding tendency. Patient satisfaction was very high for both types of prosthetic rehabilitation. The FFP group scored only slightly better for chewing comfort and general satisfaction. CONCLUSION: These data indicate that from the clinical and microbiological standpoint, as well as patient satisfaction, both an OD and a FFP offer a favourable long-term outcome.

The impact of periodontal therapy and the adjunctive effect of antiseptics on breath odor-related outcome variables: a double-blind randomized study.

BACKGROUND: Bad breath is often caused by periodontitis and/or tongue coating. This study followed the impact of initial periodontal therapy on several halitosis-related outcome variables over a 6-month period. Organoleptic ratings are often uncomfortable for the patient and have several disadvantages. They are, for instance, influenced by external parameters (e.g., food intake and cosmetics) and need to be calibrated among researchers worldwide. A second aim was to evaluate the reliability of saliva incubation as an in vitro indirect test for breath recording. METHODS: In this double-blind, randomized, medium-term, parallel study 45 moderate periodontitis patients without obvious tongue coating were enrolled. Besides a one-stage, full-mouth disinfection and oral hygiene improvement (including daily tongue scraping), patients were instructed to rinse daily for 6 months with one of the following products (randomly allocated): chlorhexidine (CHX) 0.2% + alcohol, CHX 0.05% + cetyl pyridinium chloride (CPC) 0.05% without alcohol (a new formulation), or a placebo solution. At baseline and 3 and 6 months, a series of parameters were recorded including: concentration of volatile sulfide compounds (VSC), tongue coating, and an estimation of the microbial load (at anterior and posterior parts of the tongue, saliva, dental plaque). The intraoral VSC ratings were compared to in vitro VSC recordings and organoleptic evaluations of the headspace air from 1 and 2 hours incubated saliva (0.5 ml, 37 degrees C, anaerobic chamber). RESULTS: Even though the initial VSC values were not high (+/-90 ppb with only 18 patients revealing more than 100 ppb), significant (P <0.05) reductions could be achieved in the CHX and CHX + CPC group, and to a lower extent in the placebo group (P = 0.10). Tongue scraping resulted in a significant reduction (P < or =0.05) of the tongue coating up to month 6 in the placebo and CHX + CPC group, but not in the CHX group (confusion due to staining). The CHX and CHX + CPC group showed, in comparison to baseline, significant (P <0.001) reductions in the number of anaerobic species in the supragingival plaque, in the saliva, and on the anterior part of the tongue. For the posterior part of the tongue the microbial changes remained < or =0.3 log values (P >0.05). For the placebo group, the microbial changes never reached a level of significance (< or =0.3 log values). A strong correlation was found between the intraoral VSC ratings and the 1-hour (r = 0.48, P <0.0001; r = 0.54, P = 0.0003 for baseline data only) and 2-hour (r = 0.43, P <0.0001) VSC production of incubated saliva. The latter also correlated very strongly (r = 0.71) with the number of anaerobic species in the saliva. The VSC values and organoleptic ratings of the incubated saliva also correlated strongly with each other (r = 0.64 for 1-hour and 0.73 for 2-hour incubation). CONCLUSIONS: The results of this study indicate that in patients with moderate periodontitis, initial periodontal therapy including tongue scraping did not have a significant effect on the microbial load of the tongue and had only a weak impact on the VSC level, except when combined with a mouthrinse. Saliva incubation can be used as an indirect way to score breath odor. It offers simplicity, objectivity, and is less invasive.

A 0.05% cetyl pyridinium chloride/0.05% chlorhexidine mouth rinse during maintenance phase after initial periodontal therapy.

BACKGROUND: Chlorhexidine (CHX) mouth rinse/spray can still be considered the gold standard in the chemical prevention of plaque formation and development of gingivitis. The product unfortunately has some side effects, such as extrinsic tooth staining, poor taste, taste disturbance, sensitivity changes in tongue, pain and irritation because of the alcohol content. These side effects led to the search of new formulations. METHODS: In this double-blind, randomized, long-term, parallel study, 48 moderate periodontitis patients rinsed for 6 months (starting immediately after a "one-stage, full-mouth" disinfection) with one of the following products: CHX 0.2%+alcohol (Corsodyl), CHX 0.05%+ cetyl pyridinium chloride (CPC) 0.05% and no alcohol (Perio-Aid Maintenance, a new formulation), or the placebo of the latter. After 1, 3 and 6 months a series of clinical and microbiological parameters were recorded for the supra- and subgingival area as well as for saliva. RESULTS: Although there was a significant treatment impact (mechanical debridement) in all groups, both CHX solutions further decreased both plaque and gingivitis indices (p<0.001 and p<0.05, respectively), when compared with placebo. This was also reflected by additional reductions in the number of CFU/ml of aerobic and especially anaerobic species and by a suppression of Streptococcus mutans (versus an overgrowth for the placebo), in all niches. Differences between both CHX solutions were never encountered. The subjective ratings were slightly in favour of the new CHX-CPC formulation when compared with the other CHX-alcohol formulation, especially for taste of the product (p<0.05), but less impressive for the staining of teeth and tongue. CONCLUSIONS: The results of this study demonstrated the potential of a new CHX 0.05%+CPC 0.05% non-alcoholic formulation as an effective antiplaque agent for long-term use with reduced subjective side effects.

A salivary incubation test for evaluation of oral malodor: a pilot study.

BACKGROUND: Breath odor is scored by different techniques, each with its own shortcomings. Organoleptic ratings are uncomfortable for the patient, subjective, influenced by external parameters including food and cosmetics, and especially lack international calibration. Portable sulphide monitors are relatively expensive and neglect several major malodorous molecules (e.g., butyric and propionic acids, putrescine, and cadaverine). Gas chromatography necessitates expensive devices and experienced technicians. This pilot study explored the applicability of a new technique (saliva incubation) by comparing its discrimination power, in a morning bad breath inhibition study of antiseptics, to those of hydrogen sulphide (H2S) measurement devices and organoleptic ratings. METHODS: After a professional cleaning, 8 periodontally healthy students abstained from all means of mechanical plaque control for 5 experimental periods of 7 days, with intervening washout periods of at least 2 weeks. During each experimental period, the students rinsed only twice daily with different antiseptics. At day 7, morning breath was scored clinically (volatile sulphide compound [VSC] level and organoleptic ratings), and 1.5 ml of saliva was collected and divided between 3 glass tubes that were sealed and incubated (37 degrees C, anaerobic chamber). Immediately after collection and after 3 and 6 hours of incubation, the headspace air in one of the tubes was examined for VSC production and organoleptic measurements. RESULTS: The investigations of the incubated saliva correlated well with the 7-day intraoral VSC recordings and organoleptic ratings (P < or = 0.005). Moreover, evaluations showed a similar interproduct ranking for their efficacy in malodor control. The power analyses indicated a higher discrimination power for the saliva incubation test than for the intraoral registrations. CONCLUSIONS: The strong correlation between odor production of incubated saliva and clinical assessments suggests that the saliva incubation test may be used as an indirect method to measure oral malodor and can be employed to investigate the antimalodor effectiveness of oral hygiene products.

Adhesion of Porphyromonas gingivalis serotypes to pocket epithelium.

BACKGROUND: Porphyromonas gingivalis, a key pathogen in periodontitis, is able to adhere to and invade the pocket epithelium. Different capsular antigens of P. gingivalis have been identified (K-serotyping). These P. gingivalis capsular types show differences in adhesion capacity to human cell lines or to cells cultured on a feeder layer or stromal equivalent. METHODS: The adhesion capacity of different P. gingivalis serotypes (6 capsular types and non-encapsulated strains) was compared on in vitro cultured epithelial monolayers from periodontal pockets of patients with periodontitis. The degree of adherence of P. gingivalis was evaluated by both culture and fluorescence microscopy. RESULTS: Non-encapsulated strains adhered significantly more than their capsulated variants. Capsule type 4 (K-4) adhered slightly better than the remaining K-types. CONCLUSION: This study indicates that the presence and type of capsule have a significant influence on the initial adhesion of P. gingivalis to human periodontal pocket epithelial cells.

Can toothpaste or a toothbrush with antibacterial tufts prevent toothbrush contamination?

BACKGROUND: Periopathogens and cariogenic species survive more than 48 hours on toothbrushes even under dry storage. This study examined a decontamination of toothbrushes by means of the bactericidal effect of different toothpastes or a special coating of the tufts. METHODS: Eight untreated periodontitis patients were professionally brushed, using a new toothbrush per quadrant and 3 different toothpastes containing either amine/stannous fluoride (AmF/SnF2), amine fluoride (AmF), or a mild surfactant as major antibacterial component (excipient), or no paste (C). The brushes were rinsed and stored dry at room temperature. At different time intervals, 4 tufts per toothbrush were processed for selective and non-selective culturing procedures. The protocol was repeated comparing AmF/SnF2 and C with 2 toothbrushes with coated tufts (coat1 and coat2). RESULTS: At baseline, C brushes (n = 16) harbored 10(7), 10(8), and 10(6) colony forming units (CFU) of aerobic, anaerobic, and black pigmented species, respectively. After 24 hours 10(6) CFU of aerobic and anaerobic species could still be cultured from 14 of the 16 brushes. Black-pigmented species remained detectable up to 4 hours; detection frequencies for Fusobacterium nucleatum and Streptococcus mutans at 24 hours were 5/16. With AmF/SnF2 and AmF toothpaste the number of adhering bacteria was already 2 log lower at baseline (P<0.05), but not for the excipient (0.5 log, P = 0.7). With AmF/SnF2 no vital species could be detected after 8 hours. AmF reduced the bacterial survival rate significantly more than the excipient, but less than AmF/SnF2 (P<0.05). The bactericidal effect of coat1 and coat2 was negligible when compared to C (P>0.20), and clearly inferior to the AmF/SnF2 paste (P<0.0001). After 24 hours these brushes still harbored >10(5) CFU. CONCLUSIONS: Toothpastes can significantly reduce contamination of toothbrushes, but their bactericidal activity is dependent on their composition. Coated tufts failed to limit the bacterial contamination.