Microbial communities of titanium versus zirconia abutments on implant-supported restorations: Biodiversity composition and its impact on clinical parameters over a 3-year longitudinal prospective study.

BACKGROUND: Shifts in microbial communities are common over time, but they may disturb the host-microbiome homeostasis and result in inflammation of the peri-implant issues if a dysbiotic biofilm is established. PURPOSE: Considering that different oral substrate surfaces may have a relevant impact on the microbial adhesion and colonization, the aim of this study was to investigate the microbial communities of the biofilm formed on single-implant restorations using titanium or zirconia abutments and how they correlate with clinical parameters after 3-years of implant loading. MATERIALS AND METHODS: MiSeq sequencing of 16S rRNA amplicons was used to characterize the oral biofilms of individuals (n = 20) who were sampled longitudinally during 3 years of masticatory loading. Bioinformatics analysis and multivariate statistical analysis were used to evaluate the microbial diversity and clinical outcomes. RESULTS: Microbiomes of both abutment materials presented high alpha-diversity indices during all the experimental period, irrespective of the time of sampling. Microbial communities of titanium and zirconia were quite different over time, differing about 30% after 3 years of functional loading. Similarity of microbiomes between tested abutments and contralateral teeth was also low, ranging between 45% and 50% after 3 years of investigation. Periodontal pathogens commonly associated with peri-implantitis were found in both groups. Furthermore, both abutment materials presented strong correlations of diversity indices and microbial taxa with clinical outcomes. CONCLUSIONS: The type of abutment substrate significantly influenced diversity and clustering of communities during 3 years of functional loading. The time of sampling had no effect on the variables. Large correlations were found between microbial findings and clinical outcomes.

Oral bacterial colonization on dental implants restored with titanium or zirconia abutments: 6-month follow-up.

OBJECTIVE: This investigation aimed to characterize in a 6-month follow-up the microbial profile of implants restored with either titanium or zirconia abutments at the genus or higher taxonomic levels. METHODS: Twenty healthy individuals indicative for implant-retained single restorations were investigated. Half of participants were restored with titanium and half with zirconia abutments. Biofilm was collected from the implant-related sites after 1, 3, and 6 months of loading. The 16S rDNA genes were amplified and sequenced with Roche/454 platform. RESULTS: A total of 596 species were identified in 360 samples and grouped in 18 phyla and 104 genera. Titanium- or zirconia-related sites as well as teeth showed similar total numbers of operational taxonomic units (OTUs) colonizing surfaces over time. Firmicutes, Proteobacteria, Fusobacteria, Bacteroidetes, and Actinobacteria were the most prevalent phyla with significant differences between different surfaces and time point. Unclassified genera were found in lower levels (1.71% up to 9.57%) on titanium and zirconia samples when compared with teeth, with no significant differences. CONCLUSION: Titanium- and zirconia-related surfaces are promptly colonized by a bacterial community similar to those found in the remaining adjacent teeth. Results suggest a selective adhesion of different bacterial genotypes for either titanium or zirconia surfaces. Data also indicate a significant interaction between the relative effects taxa, time point, and sampling site. CLINICAL RELEVANCE: The present study disclosed a wider spectrum of microorganisms colonizing either titanium- or zirconia-related microbiomes in very early stage of implant colonization, revealing differences and suggesting a probably specific mechanism for selective bacterial adhesion.

Microbial diversity of the supra- and subgingival biofilm of healthy individuals after brushing with chlorhexidine- or silver-coated toothbrush bristles.

Nanoparticulate silver has recently been reported as an effective antimicrobial agent. The aim of this clinical study was to investigate the potential changes on the oral microbiota of healthy individuals after controlled brushing with chlorhexidine- or silver-coated toothbrush bristles. Twenty-four healthy participants were enrolled in this investigation and randomly submitted to 3 interventions. All the participants received, in a crossover format, the following toothbrushing interventions: (i) chlorhexidine-coated bristles, (ii) silver-coated bristles, and (iii) conventional toothbrush (Control). All the interventions had a duration of 30 days. The DNA checkerboard hybridization method was used to identify and quantify up to 43 microbial species colonizing the supra- and subgingival biofilm. The supragingival samples presented higher genome counts than the subgingival samples (p < 0.0001). The total genome counts from the Control group showed the highest values, followed by the silver and chlorhexidine groups (p < 0.0001). After 4 weeks of brushing, the silver-coated and chlorhexidine-coated bristles were capable of reducing or maintaining lower levels of the bacterial counts of the putative periodontal pathogens Tanerella forsythia, Treponema denticola, and Porphyromonas gingivalis. Other major periodontal pathogens, such as Prevotella intermedia, Fusobacterium nucleatum, Prevotella nigrescens, and Parvimonas micra, were also detected at lower levels. The toothbrush bristles impregnated with silver nanoparticles reduced the total and individual genome count in the supra- and subgingival biofilm after 4 weeks of brushing. Chlorhexidine was not effective in reducing the total genome counts in both supra- or subgingival biofilm after 4 weeks of brushing. Chlorhexidine reduced the individual genome counts in the supragingival biofilm for most of the target species, including putative periodontal pathogens.

Bacterial adhesion on the titanium and zirconia abutment surfaces.

OBJECTIVE: Microorganisms harboring the oral cavity, mainly those related to periodontal diseases, are the most potential etiologic factor of failure in long-term implant treatment. The material used for abutment components may influence the adhesion and colonization of microbial species. The aim of this in vivo investigation was to evaluate the biofilm formation on machined (MPT) or cast titanium (CPT) and zirconia abutments (Zc). METHODS: Six healthy subjects were enrolled in this randomized crossover clinical investigation. The study was conducted in three phases according to abutment surface evaluated. Each subject used an individual oral splint containing four disks of the same tested substrate, two located in the anterior and two in the posterior region, totalizing 12 specimens for subject. Participants were asked to use the removable intraoral splint during 24 h. DNA checkerboard hybridization method was used to identify and quantify 38 bacterial species colonizing formed biofilm on the abutment substrates. RESULTS: Pathogens and non-pathogens species were found colonizing the three substrates surfaces. Fusobacterium nucleatum, Neisseria mucosa, Porphyromonas aeruginosa, Peptostreptococcus anaerobios, Staphylococcus aureus, Streptococcus gordonii, Streptococcus parasanguinis, and Tanerella forsythia were the only species with no significant differences over the tested materials (P > 0.05). All the other target species presented significant differences sought by Friedman test (P < 0.0001). CONCLUSIONS: There was a significant difference in the total bacterial count between the three groups. CPT presented the higher mean counts, followed by MPT and Zc. CPT group also showed a higher mean incidence of species than MPT and Zc. The anterior or posterior region of disks placement did not show significant differences in relation to bacterial adhesion.

In vivo evaluation of Candida spp. adhesion on titanium or zirconia abutment surfaces.

OBJECTIVE: Candida spp. have been found colonising implant sites in healthy or diseased subjects. The aim of this in vivo study was to evaluate the Candida spp. adhesion on machined or cast titanium and zirconia (Zc) abutment substrates. DESIGN: Six healthy subjects were enrolled in this randomised crossover clinical investigation. The study was conducted in three phases according to evaluated substratum. Participants were advised to use an intraoral splint containing four discs of the same tested substrate for 24h. Two discs were located in the anterior region and two in the posterior region. DNA checkerboard hybridisation method was used to detect and quantify five different Candida species. Data on the surface roughness and the total area of discs covered by formed biofilm were also provided to correlate the species and biofilm found between different substrates. RESULTS: Zc presented the highest means of surface roughness. Total area of the biofilm covering was not different in the tested groups. Moderate to high levels of target microorganisms were recorded for all the tested substrates. Zc showed the lowest indices, followed by machined pure titanium (MPT) and cast and polished titanium (CPT). Candida albicans and Candida krusei were not detected in the Zc group. The region of disc placement did not show differences in relation to Candida adhesion. CONCLUSIONS: There was a significant difference in the total cell count between the three groups. CPT presented the higher mean counts, followed by MPT and Zc. There was no positive correlation between the cell counts recorded and the surface roughness or total area of formed biofilm.

Assessing the oral microbiota of healthy and alcohol-treated rats using whole-genome DNA probes from human bacteria.

OBJECTIVE: This study aimed to evaluate the capacity of whole-genome DNA probes prepared from human oral bacteria to cross-react with bacteria from the oral cavity of rats, and to assess the influence of alcohol ingestion on the animals' oral biofilm. DESIGN: Twenty four mature Wistar rats were equally divided in two groups. One group (control) was fed balanced diet of rat pellets and water. The alcohol-treated group (AT) received the same diet and 20% ethanol solution. Upon euthanasia after 30 days, bacterial samples from the oral biofilm covering the animals' teeth were collected using microbrushes. Bacteria identification and quantification were performed using the DNA checkerboard hybridization method with 33 probes prepared from human oral bacteria. Signals corresponding to bacterial genome counts and percentages were compared using a Mann-Whitney U test with a significance level <0.05. RESULTS: Cross-reaction for all targeted species, except Streptococcus mutans and Streptococcus mitis-like species, occurred in the control group. Escherichia coli, Pseudomonas aeruginosa, Porphyromonas endodontalis, and Veillonella parvula-like species only produced detectable signals in the AT group. Significantly more signals were detected in the control group compared to the AT group (p=0.001). The percentage of E. coli-like species was highest in both groups. CONCLUSIONS: Whole-genome DNA probes prepared from human oral bacteria can cross-react with rats' oral bacterial species. Alcohol consumption is associated with lower levels and diversity of bacterial species in the oral cavity of rats.

Effect of electromagnetic field on bone regeneration around dental implants after immediate placement in the dog mandible: a pilot study.

BACKGROUND: Accelerating bone healing around dental implants can reduce the long-term period between the insertion of implants and functional rehabilitation. OBJECTIVE: This in vivo study evaluated the effect of a constant electromagnetic field (CEF) on bone healing around dental implants in dogs. MATERIALS AND METHODS: Eight dental implants were placed immediately after extraction of the first pre-molar and molar teeth on the mandible of two male dogs and divided into experimental (CEF) and control groups. A CEF at magnetic intensity of 0.8 mT with a pulse width of 25 µs and frequency of 1.5 MHz was applied on the implants for 20 min per day for 2 weeks. RESULT AND CONCLUSION: After qualitative histological analysis, a small quantity of newly formed bone was observed in the gap between the implant surface and alveolar bone in both groups.

Shear bond strength between Ni-Cr alloy bonded to a ceramic substrate.

INTRODUCTION: The aim of this study was to evaluate the shear bond strength between a Ni-Cr alloy and a ceramic system submitted or not to thermocycling. MATERIALS AND METHODS: Forty-eight cylinder blocks of Ni-Cr with 3.0 mm diameter by 4.0 mm hight and 48 disc-shaped specimens (7.0 mm in diameter by 2.0 mm thick) composed of ceramic were prepared. The Ni-Cr cylinder blocks were randomised in two groups of 24 specimens each. One group was submitted to air-particle abrasion (sandblasting) with 50 µm Al(2)O(3) (0.4-0.7 MPa) during 20 s, and the other group was submitted to mechanical retentions with carbide burrs. Each group was subdivided into other two groups (n = 12), submitted or not to thermocycling (500 cycles, 5-55°C). The cylinder blocks were bonded to the disc-shaped ceramic specimens under 10 N of load. The shear bond strengths (MPa) were measured using a universal testing machine at a cross head speed of 0.5 mm/min and 200 kgf of load. The data were submitted to statistical analysis (anova and Tukey's test). RESULTS: The air-particle abrasion group exhibited significantly higher shear bond strength when compared to drilled group (p < 0.05). CONCLUSIONS: Thermocycling decreased significantly the bond strengths for all groups tested.

Use of checkerboard DNA-DNA hybridization to evaluate the internal contamination of dental implants and comparison of bacterial leakage with cast or pre-machined abutments.

AIMS: To evaluate the checkerboard DNA-DNA hybridization method for detection and quantitation of bacteria from the internal parts of dental implants and to compare bacterial leakage from implants connected either to cast or to pre-machined abutments. MATERIALS AND METHODS: Nine plastic abutments cast in a Ni-Cr alloy and nine pre-machined Co-Cr alloy abutments with plastic sleeves cast in Ni-Cr were connected to Branemark-compatible implants. A group of nine implants was used as control. The implants were inoculated with 3 microl of a solution containing 10(8) cells/ml of Streptococcus sobrinus. Bacterial samples were immediately collected from the control implants while assemblies were completely immersed in 5 ml of sterile Tripty Soy Broth (TSB) medium. After 14 days of anaerobic incubation, occurrence of leakage at the implant-abutment interface was evaluated by assessing contamination of the TSB medium. Internal contamination of the implants was evaluated with the checkerboard DNA-DNA hybridization method. RESULTS: DNA-DNA hybridization was sensitive enough to detect and quantify the microorganism from the internal parts of the implants. No differences in leakage and in internal contamination were found between cast and pre-machined abutments. Bacterial scores in the control group were significantly higher than in the other groups (P<0.05). CONCLUSION: Bacterial leakage through the implant-abutment interface does not significantly differ when cast or pre-machined abutments are used. The checkerboard DNA-DNA hybridization technique is suitable for the evaluation of the internal contamination of dental implants although further studies are necessary to validate the use of computational methods for the improvement of the test accuracy.