Bacterial response to Ti-35Nb-7Zr-5Ta alloy incorporated with calcium, phosphate and magnesium.

High implant survival rates have been achieved in recent decades due to continual modifications in implant design and surface topography, however there is still an ongoing quest to control peri-implant bone loss. The objective of this work was to develop Ti-35Nb-7Zr-5Ta (TNZT) alloys, perform physicochemical and morphological characterization of their surface modified by electrolytic oxidative plasma technique with ions related to osseointegration and lastly evaluate bacterial colonization in vitro. Three groups were evaluated: C group (polished TNZT), CaP group (sodium ß glycerophosphate + calcium acetate) and Mg group (magnesium acetate). Before and after anodizing the surfaces, physicochemical and morphological analyses were performed: scanning electron microscopy with field emission gun (FEG-SEM), energy dispersion spectroscopy (EDS), X-ray diffraction (DRX), wettability (goniometer) and roughness (rugometer). Controlled and treated specimens were contaminated with unstimulated saliva collected from 10 healthy volunteers. Then, biofilm samples were collected and up to 35 microbial species, including commensal and pathogenic microorganisms, were identified and quantified by the Checkerboard DNA-DNA Hybridization method. The CaP group modified the surface morphology in the form of pores, while the Mg group modified it in the form of flakes. The contact angle was significantly smaller in the CaP group. The average roughness was higher in the CaP and Mg groups. A smaller total amount of bacteria was identified in the Mg group and relevant differences were found in the microbial profile associated with different surface treatments. Therefore, considering the microbiological profile and for the prevention of peri-implantitis, the Mg group presented more satisfactory and encouraging results for the manufacture of dental implants.

Self-ligating brackets exhibit accumulation of high levels of periodontopathogens in gingival crevicular fluid.

Different types of brackets seem to influence the disruption of the oral microbial environment. Therefore, the aim of this study was to evaluate the influence of self-ligating brackets on the gingival crevicular fluid levels of the putative periodontal pathogens Aggregatibacter actinomycetemcomitans sorotype a (Aaa), Tannerella forsythia, Fusobacterium nucleatum, and Porphyromonas gingivalis. Sixty samples of crevicular fluid of twenty patients (11 boys and 9 girls) were analysed at baseline (T0) and after 30 (T1) and 60 (T2) days of bonding of the self-ligating (In-Ovation®R, Dentsply, GAC or SmartClip™, 3 M Unitek, Monrovia, CA, USA) and of one conventional bracket (Gemini™, 3 M Unitek, Monrovia, CA, USA) used with elastomeric ligatures. Total DNA from samples was extracted using CTAB-DNA precipitation method and Real-time PCR was performed to analyse bacterial level. Non-parametric Friedman and Wilcoxon tests were used for data analysis (p value of < 0.05). F. nucleatum presented a different level among the different brackets at T1 (p = 0.025), the highest level in the Gemini™ bracket when compared to the SmartClip™ bracket (p = 0.043). P. ginigvalis levels increased in the In-Ovation®R (p = 0.028) at T1. The subgingival levels of bacterial species associated with periodontal disease P. ginigvalis increased in the self-ligating brackets In-Ovation®R.Clinical Relevance: Some kinds of brackets could provide more retentive sites than others, and it seems to modulate the subgingival microbiota, since, in this study, we could observe the increase of the species associated with periodontal disease. Preventive protocols should be adopted in the use of self-ligating brackets.

Brushing associated with oral irrigation in maintaining implants and overdentures hygiene - a randomized clinical trial.

Evaluate, through a randomized clinical trial, the efficacy of brushing associated with oral irrigation in maintaining implant and overdenture hygiene. Thirty-eight participants, who had a clinically acceptable conventional maxillary complete denture and mandibular overdenture retained by either implants or mini-implants using an O-ring-retained system, were enrolled to participate in the study. They were instructed to use two different hygiene methods, in a random sequence for a period of 14 days, with a 7-day wash-out interposed period: (I) mechanical brushing (MB); (II) association of mechanical brushing with oral irrigation (WP). Biofilms from both subgingival sulci and overdentures were collected and processed by Checkerboard DNA-DNA hybridization method at baseline and after using the proposed hygiene protocols. Comparisons were performed using Wilcoxon test and Friedman test with Benjamini-Hochberg false discovery rate, followed by Conover post-hoc test (α = 0.05). In the subgingival sulci-related biofilm, a lower number of microbial cells were detected, after WP compared to the MB method (P < 0.001). The findings of overdenture-related biofilm suggest that both methods were similar (P = 0.607) being the identified microbiota qualitatively coincident after each method. Despite the number of microbial counts, it was concluded that the association of mechanical brushing with oral irrigation was more effective in reducing microorganisms in the subgingival sulci biofilm; however, the same outcome was not observed in the overdentures.

Comparison of oral microbiome profile of polymers modified with silver and vanadium base nanomaterial by next-generation sequencing.

This study aimed to evaluate two methods of the incorporation of nanostructured silver vanadate decorated with silver nanoparticles (AgVO3) into acrylic resin and characterize the profile of early and late microbial communities in class and family taxonomic level by pyrosequencing. The specimens were made by adding different concentrations of AgVO3 (1, 2.5, and 5%) to the heat-activated acrylic resin by two methods: vacuum spatulation (VS) and polymeric film (PF). A control group (0%) without AgVO3 was also obtained for both methods. After 24 h and 7 days of incubation in human saliva, biofilm samples were collected, DNA was extracted, and 16S rRNA genes were sequenced by the 454-Roche sequencing platform. Seventeen classes and 51 families of bacteria were identified. The abundance of Bacteroidia, Bacilli, Negativicutes, Fusobacteria and Betaproteobacteria classes decreased after 7 days of incubation, and Clostridia, Gammaproteobacteria, and unclassified bacteria increased. The Negativicutes and Betaproteobacteria classes were more abundant when the PF method was used, and Gammaproteobacteria was more abundant when VS was used. The incorporation of 5% AgVO3 promoted a reduction in the prevalence of Bacilli, Clostridia, Negativicutes, Betaproteobacteria, and unclassified bacteria, and increased Gammaproteobacteria. The addition of AgVO3 to acrylic resin altered the early and mature microbiome formed on the specimen surface, and the PF method presented a more favorable microbial profile than the VS method.

Genome analysis and clinical implications of the bacterial communities in early biofilm formation on dental implants restored with titanium or zirconia abutments.

This cross-sectional study aimed to identify and quantify up to 42 target species colonizing the early biofilm of dental implants restored with titanium or zirconia abutments. A total of 720 samples from 20 healthy individuals were investigated. Biofilm samples were collected from the peri-implant sulci, inner parts of implants, abutment surfaces and prosthetic crowns over a functioning period of 30 days. Checkerboard DNA-DNA hybridization was used for microbial detection and quantitation. Clinical characteristics (probing depth, bleeding on probing, clinical attachment level and marginal bone loss) were also investigated during the monitoring period. Genome counts were low at the implant loading time point for both the abutment materials, and increased over time. Both the titanium and the zirconia groups presented similar microbial counts and diversity over time, and the microbiota was very similar to that colonizing the remaining teeth. Clinical findings were consistent with a healthy condition with no significant difference regarding marginal bone loss between the two materials.

Genomic identification of microbial species adhering to maxillofacial prostheses and susceptibility to different hygiene protocols.

This study investigated the microbial colonization of maxillofacial prostheses and support tissues using the Checkerboard DNA-DNA hybridization method, and the efficacy of 0.12% chlorhexidine gluconate, 10% Ricinus communis solutions, or brushing, on colony forming unit (CFU) reduction in monospecies biofilms (Candida glabrata, Staphylococcus aureus, Streptococcus mutans, Escherichia coli, Enterococcus faecalis, and Pseudomonas aeruginosa) formed on two silicones (MDX 4-4210 and Bio-Skin). Biofilm was harvested from 43 maxillofacial prosthesis wearers for detection of 38 species of microorganisms. The CFU counts of the six above mentioned species were recorded after using the hygiene protocols. All 38 investigated species were identified in prostheses and tissues, with a higher prevalence in the prostheses. 0.12% chlorhexidine gluconate immersion showed the greatest antimicrobial effectiveness, followed by mechanical brushing protocols. MDX 4-4210 silicone produced lower CFU counts than Bio-Skin.

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.

Identification of oral bacteria on titanium implant surfaces by 16S rDNA sequencing.

OBJECTIVE: To characterize the profile of microbial communities colonizing titanium implants with different surface treatments after exposure to the oral environment at the genus or higher taxonomic level. MATERIAL AND METHODS: Sixteen titanium disks, machined or sandblasted large-grit and acid-etched (SLA), were mounted on removable intraoral splints worn by four patients. After 24 h of intraoral exposure, biofilm samples were collected from disks and supra/subgingival teeth areas. The 16S rDNA genes from each sample were amplified, sequenced with the Miseq Illumina instrument and analyzed. RESULTS: A total of 29 genera and seven more inclusive taxa, representing the phyla Firmicutes, Proteobacteria, Fusobacteria, Bacteroidetes, Actinobacteria and candidate division TM7 were identified in both titanium surfaces and teeth. No differences were found in relation to the operational taxonomic units (OTUs) and microbial diversity, assessed by Chao 1 and Shannon indices, when comparing SLA and machined titanium surfaces. CONCLUSIONS: Machined and SLA surfaces are colonized by similar numbers of prokaryotic OTUs after 24 h of exposure to the oral environment. Higher complexity of the titanium surface topography in the initial phase of biofilm maturation does not seem to significantly influence the colonizing microbiota.

Experimental conical-head abutment screws on the microbial leakage through the implant-abutment interface: an in vitro analysis using target-specific DNA probes.

OBJECTIVES: The aim of this in vitro study was to identify and quantify up to 38 microbial species from human saliva penetrating through the implant-abutment interface in two different implant connections, external hexagon and tri-channel internal connection, both with conventional flat-head or experimental conical-head abutment screws. MATERIAL AND METHODS: Forty-eight two-part implants with external hexagon (EH; n = 24) or tri-channel internal (TI; n = 24) connections were investigated. Abutments were attached to implants with conventional flat-head or experimental conical-head screws. After saliva incubation, Checkerboard DNA-DNA hybridization was used to identify and quantify up to 38 bacterial colonizing the internal parts of the implants. Kruskal-Wallis test followed by Bonferroni's post-tests for multiple comparisons was used for statistical analysis. RESULTS: Twenty-four of thirty-eight species, including putative periodontal pathogens, were found colonizing the inner surfaces of both EH and TI implants. Peptostreptococcus anaerobios (P = 0.003), Prevotella melaninogenica (P < 0.0001), and Candida dubliniensis (P < 0.0001) presented significant differences between different groups. Means of total microbial count (×104 , ±SD) for each group were recorded as follows: G1 (0.27 ± 2.04), G2 (0 ± 0), G3 (1.81 ± 7.50), and G4 (0.35 ± 1.81). CONCLUSIONS: Differences in the geometry of implant connections and abutment screws have impacted the microbial leakage through the implant-abutment interface. Implants attached with experimental conical-head abutment screws showed lower counts of microorganisms when compared with conventional flat-head screws.

Microbiological and clinical outcomes of fixed complete-arch mandibular prostheses supported by immediate implants in individuals with history of chronic periodontitis.

OBJECTIVES: The aim of this study was to assess the microbiological and clinical outcomes of immediate implants placed in chronically infected sockets for rehabilitation with fixed full-arch mandibular prostheses. MATERIAL AND METHODS: Fourteen individuals (mean age 60.14 ± 7.69 years) were enrolled in this investigation and followed up until 8 months of function. Microbiological (microbial count and profile) and clinical (probing depth, clinical attachment level, bleeding on probing, and bone resorption) parameters were conducted before teeth extraction (T0 - baseline) and after 4 (T1 ) and 8 (T2 ) months of loading. Thirty-nine microbial species including periodontopathogenic species and Candida spp. were detected and quantified by DNA checkerboard analysis. RESULTS: Moderate to high levels of pathogenic and non-pathogenic species were found colonizing teeth and implant-related sites. No significant differences in total or individual microbial counts and microbial profile were found over time (P = 0.4929). Probing depth values from teeth (T0 : 3.05 ± 1.45) were significantly higher when compared with implants (T1 : 1.81 ± 0.56; T2 : 1.66 ± 0.53; P < 0.0001). High percentages of bleeding sites were found for both teeth and implants, with the highest values recorded for teeth (P < 0.05). No significant differences were detected comparing marginal bone resorption over time. CONCLUSIONS: Total and individual counts of target species did not differ between teeth and implants for 8 months of investigation. The mean proportions of pathogenic and non-pathogenic species remained unaltered, and no clinical complications were reported over time. Data obtained suggest that immediate loading of complete mandibular prostheses retained by implants placed immediately after extraction may be a viable treatment option for edentulous individuals with previous history of periodontal disease.

Different sealing materials preventing the microbial leakage into the screw-retained implant restorations: an in vitro analysis by DNA checkerboard hybridization.

OBJECTIVES: The aim of this controlled in vitro study was to identify and quantify up to 38 microbial species penetrating through the screw-retained implant prostheses with different sealing materials. MATERIAL AND METHODS: Sixty morse cone implants were restored with single-unit screw-retained prostheses. All the components were randomly divided into five groups (n = 12) according to the proposed materials: (1) polytetrafluoroethylene tape+composite resin; (2) polytetrafluoroethylene tape+gutta-percha; (3) polytetrafluoroethylene tape+light-polymerized provisional composite; (4) cotton pellet+gutta-percha; and (5) cotton pellet+light-polymerized provisional composite. Human saliva was used as contaminant media, and DNA checkerboard hybridization was used to identify and quantify microbial species. RESULTS: Microbial leakage was observed in all groups: M. salivarium, S. pasteuri, P. nigrescens, and P. melaninogenica were the species presenting the highest values of genome count, prevalence, and proportion within the groups. The total microbial mean counts (×105 , ±SD) were as follows: Group 1 (2.81 ± 0.38), Group 2 (3.41 ± 0.38), Group 3 (6.02 ± 1.48), Group 4 (6.40 ± 1.42), and Group 5 (17.45 ± 1.67). Group 5 showed the higher microbial counts (P < 0.001). CONCLUSIONS: Moderate to high counts of pathogenic/nonpathogenic species were detected in the inner parts of implants from all groups. The lowest values of microbial counts were recorded for polytetrafluoroethylene tape associated with composite resin or gutta-percha; cotton pellet associated with light-polymerized provisional composite presented the highest microbial counts.

Microbial complexes levels in conventional and self-ligating brackets.

OBJECTIVES: The aims were to evaluate the levels of bacterial species in saliva and in situ and to assess whether the design of brackets influences the risk of developing periodontal disease. MATERIALS AND METHODS: Twenty patients (13.3 mean age) were bonded with self-ligating brackets and a conventional bracket. Saliva was collected before bonding and 30 and 60 days after bonding. One sample of each bracket was removed 30 and 60 days after bonding. The analysis was determined by checkerboard DNA-DNA hybridization. The data was evaluated by the non-parametric test. RESULTS: A significant increase in the levels of bacterial species in the saliva occurred in 15 of the 22 analyzed species. The self-ligating brackets presented the highest incidence percentages for the orange and red complexes 60 days after bonding. In situ analyses showed different patterns according to the bracket design. The levels of Campylobacter rectus showed significant differences (p = 0.011) 60 days after bonding among the three brackets; the highest values were observed in the In-Ovation®R bracket. CONCLUSIONS: The bracket design seems to influence the levels of bacterial species involved in periodontal disease. Considering the wide variety of bacterial species, additional studies are needed to aid in the establishment of effective protocols to prevent the development of periodontal disease during orthodontic treatment. CLINICAL RELEVANCE: A dynamic alteration in the oral microbiota may lead to inflammatory reactions in the supporting soft and hard tissues. The different types of brackets interfere with bacterial adherence. Bracket design should be considered in orthodontic treatment.

Profile of bacteria colonizing the exposed bone of patients with anti-osteoclastic drug-related osteonecrosis of the jaws.

Microbial etiology for anti-osteoclastic drug-related osteonecrosis of the jaw (ARONJ) was suggested. This study investigates any link between bacteria colonizing ARONJ sites and other oral cavity sites. Microbiota samples of 10 ARONJ patients were collected from the exposed bone, adjacent teeth, contralateral teeth, and tongue. DNA checkerboard hybridization was used for microbiota analysis with 43 genomic DNA probes prepared from human oral bacterial (38) and candida (5) species, using Socransky's bacterial complexes as a guide. The frequency and the mean proportion of each bacterial species were used. Eikenella corrodens, Streptococcus constellatus, and Fusobacterium nucleatum were dominant in the ARONJ sites and detected in most teeth samples. Staphylococcus aureus was also dominant in the ARONJ sites and tongue. Significant correlations were found between the mean proportions of bacterial species colonizing adjacent teeth, contralateral teeth, and tongue (p < 0.001, R(2) > 0.69). No significant correlation (p > 0.05, R(2) < 0.025) was found between bacteria colonizing ARONJ sites and other evaluated sites. Within the study limitations, it was concluded that the primary sources of microorganisms colonizing ARONJ sites could be other sites such as teeth and tongue. The microbial profile of the necrotic bone is predominantly colonized with bacteria from Socransky's green and orange complexes, as well as with species associated with bone infections.

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.

Analgesia evaluation of 2 NSAID drugs as adjuvant in management of chronic temporomandibular disorders.

The aim of this triple-blind full-randomized clinical trial was to quantify analgesia in masticatory muscles and temporomandibular joints after occlusal splint therapy associated with the adjuvant administration of nonsteroidal anti-inflammatory drugs (NSAID) isolated or associated with other therapeutic agents. Pain relief was also recorded. Eighteen volunteers who had been suffering from chronic pain in masticatory muscles due to temporomandibular disorders were selected after anamnesis and assessment using RDC/TMD translated to Portuguese. The 3 proposed treatments were NSAID (sodium diclofenac), panacea (sodium diclofenac + carisoprodol + acetaminophen + caffeine), and a placebo. The total treatment duration was 10 days, preceded and succeeded by patients' pain assessment. A washout interval of 11 days was established between each therapy. All participants received all treatments in different moments, in a full randomized crossover methodology. The assessment of drug therapies was performed using visual analogue scale for pain on palpation followed by 11-point numerical scale to quantify pain during treatment. Statistical analysis has shown that, after 10 days of treatment, all therapies were effective for pain relief. NSAID therapy promoted analgesia on the third day, while placebo only promoted analgesia in the eighth day. It has been concluded that sodium diclofenac used as splint adjuvant therapy, promotes significant analgesia in a shorter time.

Effectiveness of three antimicrobial mouthrinses on the disinfection of toothbrushes stored in closed containers: a randomized clinical investigation by DNA Checkerboard and Culture.

OBJECTIVES: The aim of this in vivo study was to evaluate the efficacy of three antimicrobial solutions on the disinfection of toothbrushes after storage in closed containers. MATERIALS AND METHODS: Sixteen healthy subjects were enrolled in this randomized cross-over clinical investigation. The study was conducted in four phases, in which mouthrinses (chlorhexidine gluconate-based or cetilpiridinium-based) and sterile tap water (control group) were used to individually store used toothbrushes in closed containers during 7 days of toothbrushing. Five toothbrushes were used as negative control for bacterial colonisation before contact with oral cavity. Conventional culture and DNA Checkerboard hybridization were used to detect bacterial contamination on the toothbrushes. Subsequently, the number of bacterial species on the bristles was estimated by the DNA Checkerboard method. RESULTS: One toothbrush presented bacterial contamination in the negative control test. Both culture and DNA Checkerboard showed positive signals of bacterial contamination in the toothbrushes with no differences in the frequency of detection. The control group showed higher total bacterial counts when compared with the mouthrinse groups. Porphyromonas gingivalis had the highest bacterial count followed by Parvimonas micra. CONCLUSION: Culture and DNA Checkerboard showed positive signals of bacterial contamination. Mouthrinses that contains 0.12% of chlorhexidine gluconate were more effective in reducing bacterial colonisation on the toothbrushes.

Bacterial Detection, Deformation, and Torque Loss on Dental Implants with Different Tapered Connections Compared with External Hexagon Connection after Thermomechanical Cycling.

The relationship between bacterial infiltration and internal conical Implant-Abutment Interfaces (IAIs) with different conicities still requires investigations that can offer valuable information in the clinical understanding of peri-implant health. The present study aimed to verify the bacterial infiltration of two internal conical connections with an angulation of 11.5° and 16° with the external hexagonal connection as a comparative after thermomechanical cycling using saliva as a contaminant. Test (n = 10) and control (n = 3) groups were set up. Evaluations were made on torque loss, Scanning Electron Microscopy (SEM), and Micro Computerized Tomography (MicroCT) after performing 2 × 106 mechanical cycles (120 N) and 600 thermal cycles (5°-55° C) with 2 mm lateral displacement. The contents of the IAI were collected for microbiological analysis. There was a difference (p < 0.05) in torque loss of the groups tested; groups from the 16° IAI obtained a lower percentage of torque loss. All groups presented contamination and the analysis of the results shows that the microbiological profile of the IAI differs qualitatively from the profile found in the saliva used for contamination. The mechanical loading affects the microbiological profile found in the IAIs (p < 0.05). In conclusion, the IAI environment may favor a microbiological profile different from that of saliva and the thermocycling condition may alter the microbial profile found in the IAI.

Bacterial Leakage Evaluation Through DNA-DNA Checkerboard Hybridization Technique in Morse Taper Implant-Abutment Connections: An In Vitro Study.

Purpose: The objective of this in vitro study was to evaluate the activity of local gel containing metronidazole (MN) in the leakage area, which was analyzed by the DNA-DNA checkerboard hybridization method. Materials and Methods: Thirty-six sets of Morse taper/mini-pillar implants were used in this study. These implants were equally divided into the following three groups: MN gel (test group), no MN gel (negative test group), and no gel (control). The gel was prepared with metronidazole (15%). Unstimulated saliva samples were collected, transferred to a Falcon tube, and stored at 37°C. The sets were partially immersed in microtubes containing 300 µL of saliva and were incubated at 37°C ± 1°C for 7 days. Microbial infiltration was evaluated (37 bacterial species and 5 species of Candida). The results were analyzed with Wald-Type, ANOVA, and multiple comparisons analysis between groups. Results: After comparing the quantity of microorganisms, both gel-treated groups (no MN gel and MN gel) had more significant microorganism presence than the control group (P < .001), and no significant result was found between the no MN gel and MN gel groups (P > .05). Regarding the bacteria found, the most common were Aggregatibacter actinomycetemcomitans, Prevotella melaninogenica, Bacteroides fragilis, and Candida tropicalis. Conclusion: Within the limitations of this study, it was concluded that the gel containing metronidazole used in this study was not effective in preventing the infiltration of microorganisms through the Morse taper implant-abutment interface.

Bacterial leakage along the implant-abutment interface: culture and DNA Checkerboard hybridization analyses.

OBJECTIVE: Bacterial species have been found harboring the internal surface of dental implants as consequence of their failed connections. The aim of the present study was to compare the detection frequency of bacterial leakage from human saliva through the implant-abutment interface, under non-loading conditions, using either DNA Checkerboard or culture method. MATERIALS AND METHODS: Thirty dental implants with hexagonal platforms were connected to pre-machined abutments according to the manufacturers' specifications. The assemblies were individually incubated in human saliva under anaerobic conditions for 7 days at 37°C. Afterward, contents from the inner parts of the implants were collected and evaluated with either DNA Checkerboard (s = 15) or culture (n = 15). Subsequently, identification and quantitation of bacterial species from saliva and implants were carried out for the group evaluated with the DNA Checkerboard method. RESULTS: Both DNA Checkerboard and culture showed positive signals of bacterial leakage in 6 of the 15 evaluated samples. Capnocytophaga gingivalis and Streptococcus mutans were the most frequently detected species harboring the internal surface of the implants followed by Veillonella parvula. CONCLUSION: Occurrence of bacterial leakage along the implant-abutment interface is comparably detected with both DNA Checkerboard hybridization and conventional culture methods.

Effect of sample storage time on detection of hybridization signals in Checkerboard DNA-DNA hybridization.

Long-term sample storage can affect the intensity of the hybridization signals provided by molecular diagnostic methods that use chemiluminescent detection. The aim of this study was to evaluate the effect of different storage times on the hybridization signals of 13 bacterial species detected by the Checkerboard DNA-DNA hybridization method using whole-genomic DNA probes. Ninety-six subgingival biofilm samples were collected from 36 healthy subjects, and the intensity of hybridization signals was evaluated at 4 different time periods: (1) immediately after collecting (n = 24) and (2) after storage at -20 °C for 6 months (n = 24), (3) for 12 months (n = 24), and (4) for 24 months (n = 24). The intensity of hybridization signals obtained from groups 1 and 2 were significantly higher than in the other groups (p < 0.001). No differences were found between groups 1 and 2 (p > 0.05). The Checkerboard DNA-DNA hybridization method was suitable to detect hybridization signals from all groups evaluated, and the intensity of signals decreased significantly after long periods of sample storage.