Checkerboard DNA-DNA Hybridization Method Research Articles

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 β 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 implantsGraphical

Alteration of the oral microbiota may be a responsible factor, along with estrogen deficiency, by the development of larger periapical lesions.

To answer the questions: (1) Does reducing estrogen levels influence the microbial composition of the oral cavity? (2) Does the presence of periapical lesion (PL) cause changes in the oral microbiota? (3) Since estrogen deficiency alters the oral microbiota, can this be one of the factors that contribute to the increase of the PL? Thirty-six rats were divided into four groups: sham (control), ovariectomy (OVX), control with PL (Sham + PL), and OVX + PL. After 9 weeks of OVX, the lower first molars were submitted to PL induction. After 21 days, the microbiological collection of the oral cavity was performed, and the animals were euthanized. The contents were evaluated by the checkerboard DNA-DNA hybridization method, to verify the prevalence of 40 bacterial species (divided into 7 microbial complexes). The blocks containing the lower first molars were submitted to histotechnical processing and staining with hematoxylin and eosin (HE), for the measurement of the periapical lesion area. The results were submitted to ANOVA and Kruskal-Wallis tests and Tukey and Dunn post-tests, with a significance level of 5%. In conditions of estrogen deficiency, there was alteration of the oral microbiota. The OVX groups had a higher amount of bacteria compared to the SHAM group in most of the microbial complexes (p < 0.001). The animals in the control group (with or without lesion) did not present a statistically significant difference (p > 0.001) in any of the microbial complexes. The PLs in OVX animals were significantly higher compared to SHAM animals (p < 0.001). Hypoestrogenicity conditions interfere in the oral microbiota by increasing the amount of bacteria in the saliva and influencing the progression of periapical lesions. This inedited study shows that deficiency of estrogen leads to alteration of the oral microbiota.

Resolution of Nasal Sinus Tract after Endodontic Therapy: A Case Report with Microbial Analysis

This case report describes the resolution of a 20-year misdiagnosed nasal sinus tract after root canal therapy with multiple sessions of calcium hydroxide (Ca[OH]2) intracanal medication. Clinical evaluation, including diagnostic testing and sinus tract tracing, was performed followed by a cone-beam computed tomographic scan and 3-dimensional reconstruction of the apical lesion. Bacteria and endotoxin analyses were performed from the nasal sinus tract and paired root canal infection before (s1) and after instrumentation (s2) and after 7 (s3), 14 (s4), and 21 (s5) days of Ca(OH)2 medication. The bacteria analysis was performed using the checkerboard DNA-DNA hybridization method and endotoxin quantified by the limulus amebocyte lysate method. A similar microbiota profile was found in the sinus tract and paired root canal infection. No target bacterial species were detected in the root canal at s2, s3, and s5. In contrast, Actinomyces israellii and Eubacterium nodatum were detected at s4. Differences in bacterial detection were found between s1 × s2, s3 × s4, and s4 × s5 (all P < .05). Endotoxin was detected in the root canal at all sampling times. Differences in the levels of endotoxin were found between s1 × s2, s2 × s3, and s3 × s4 (all P < .05).The bacterial analysis revealed similar microbiota profiles present in the nasal sinus tract and paired root canal infection with the participation of a wide variety of gram-positive and -negative species. Additionally, root canal therapy with multiple sessions of Ca(OH)2 intracanal medication for 21 days was effective in disinfecting the root canal system and resolving the nasal sinus tract.

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 14days, 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 andFriedman 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.

Microbial Profile and Endotoxin Levels in Primary Periodontal Lesions with Secondary Endodontic Involvement.

This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Antimicrobial Efficacy of Materials Used for Sealing the Implant Abutment Screw Hole: An In Vitro Evaluation.

The aim of this in vitro study was to assess the antimicrobial efficacy of materials (gutta percha [GP], polytetrafluoroethylene [PTFE] tape, and vinyl polysiloxane [VP]) used for sealing the implant abutment screw hole (IASH). Sixty implants with a Morse cone connection were used. The implants were randomly divided into 3 groups (20 implants/group). In groups 1, 2, and 3, the IASH was sealed with GP, PTFE, and VP, respectively. Unstimulated whole saliva (UWS) was used as a contaminant medium for the incubation of the specimens. After 7 days of incubation, the specimens were opened under aseptic conditions, and the contents of the inner parts of the IASH were collected using sterile microbrushes. The occurrence of microbial leakage through the screw access channel interface was assessed with the checkerboard DNA-DNA hybridization method. P < 0.05 was considered statistically significant. Among samples in groups 1, 2, and 3, the UWS collected before incubation showed no statistically significant differences in relation to the prevalence of species investigated. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Candida albicans were isolated from group 1 than groups 2 and 3. The choice of material for sealing the IASH is entirely dependent on the operators' choice and experience.

Evaluation of oral microbiota in undernourished and eutrophic children using checkerboard DNA-DNA hybridization

The aim of this study was to evaluate the relationship among nutritional status, gingival health and the composition of oral microbiota in children of a public school from a very poor area of San Miguel de Tucuman. Forty-five children ranging in age from 6 to 14 years old, 13 males and 32 females were studied. Twenty of these children were undernourished (Lejarraga-Morasso Table) and twenty-five were eutrophic. A clinical study that included DMF and dmf indexes, Löe Silness Plaque Index and bleeding on probing was performed. For microbiological study, saliva samples without stimulation were taken; aliquots of them were immediately placed in TAE buffer pH 7.6, adding NaOH (N and keeping at -70 °C until processed by checkerboard DNA-DNA hybridization method to check the presence of 40 oral microorganism species. Positive bleeding on probing was present in more than 80% of children, without significant differences between eutrophic and undernourished groups. Same result were obtain for the other clinical indexes (p > 0.05, Two Way ANOVA). Significant differences were found for some oral microorganism species, with a higher percentage of undernourished children harboring them. That was the case of S. gordonii (p < 0.05), Capnocitophaga gingivalis and C. ochraceae (p < 0.01 and p < 0.10, respectively), F. nucleatum ss nucleatum (p < 0.05), P. nigrescens (p < 0.10), Campylobacter gracilis (p < 0,05), and T. denticola (p < 0.10, multiple logistic regression). Significant differences were also found between children groups for E. saborreum (p < 0.001), P. acnes (p < 0.10), G. morbillorum (p < 0.05) and L. buccalis (p < 0.10). Gingivitis and bleeding on probing would not be related to nutritional status in the groups of children studied. There were significant differences for the presence of some of the main periodontal pathogen species between eutrophic and undernourished children. It would be important to study the meaning of significant differences found for the other microorganisms more deeply.

Cytokine expression in peri‐implant crevicular fluid in relation to bacterial presence

The aim was to assess clinical inflammatory parameters, cytokine levels and bacterial counts in samples from implant crevicular fluid in cases with untreated peri-implantitis. Several bacterial species known to up-regulate pro-inflammatory cytokines have been associated with peri-implantitis. The Luminex magnet bead technology was used to study cytokines in crevicular fluid. The checkerboard DNA-DNA hybridization method was used to study bacterial counts in samples from 41 implants (41 individuals). Profuse bleeding and suppuration was found in 25/41 (61.0%) of the implants. The reliability of duplicate cytokine processing was high. In the presence of profuse bleeding, higher pg/ml levels of IL-1β (p = 0.02), IL-8 (p = 0.04), TNF-α (p = 0.03) and VEGF (p = 0.004) were found. Higher concentrations of IL-1β were found in the presence of suppuration, and if Escherichia coli (p = 0.001) or Staphylococcus epidermidis (p = 0.05) could be detected. Profuse bleeding and/or suppuration in untreated peri-implantitis can be associated with higher concentrations of IL-1β, IL-8, TNF-α and VEGF in peri-implant crevicular fluid. A higher concentration of IL-1β in peri-implant crevicular fluid was found in samples that were positive for E. coli or S. epidermidis.

S-nitrosoglutathione accelerates recovery from 5-fluorouracil-induced oral mucositis.

IntroductionMucositis induced by anti-neoplastic drugs is an important, dose-limiting and costly side-effect of cancer therapy.AimTo evaluate the effect of the topical application of S-nitrosoglutathione (GSNO), a nitric oxide donor, on 5-fluorouracil (5-FU)-induced oral mucositis in hamsters.Materials and MethodsOral mucositis was induced in male hamsters by two intraperitoneal administrations of 5-FU on the first and second days of the experiment (60 and 40 mg/kg, respectively) followed by mechanical trauma on the fourth day. Animals received saline, HPMC or HPMC/GSNO (0.1, 0.5 or 2.0 mM) 1 h prior to the 5-FU injection and twice a day for 10 or 14 days. Samples of cheek pouches were harvested for: histopathological analysis, TNF-α and IL-1β levels, immunohistochemical staining for iNOS, TNF-α, IL-1β, Ki67 and TGF-β RII and a TUNEL assay. The presence and levels of 39 bacterial taxa were analyzed using the Checkerboard DNA-DNA hybridization method. The profiles of NO released from the HPMC/GSNO formulations were characterized using chemiluminescence.ResultsThe HPMC/GSNO formulations were found to provide sustained release of NO for more than 4 h at concentration-dependent rates of 14 to 80 nmol/mL/h. Treatment with HPMC/GSNO (0.5 mM) significantly reduced mucosal damage, inflammatory alterations and cell death associated with 5-FU-induced oral mucositis on day 14 but not on day 10. HPMC/GSNO administration also reversed the inhibitory effect of 5-FU on cell proliferation on day 14. In addition, we observed that the chemotherapy significantly increased the levels and/or prevalence of several bacterial species.ConclusionTopical HPMC/GSNO accelerates mucosal recovery, reduces inflammatory parameters, speeds up re-epithelization and decreases levels of periodontopathic species in mucosal ulcers.

Relation of gelsolin amyloidosis and periodontal health

Hereditary gelsolin amyloidosis (AGel amyloidosis) is a rare, dominantly inherited systemic disease with worldwide distribution, caused by a gelsolin gene mutation. We studied the periodontal conditions and microbiological plaque composition of AGel amyloidosis patients. A voluntary study group of 36 AGel amyloidosis patients (mean age 61) filled in a questionnaire. A thorough periodontal examination included periodontal pocket depth and attachment level measurements, registrations of visible plaque, bleeding on probing and panoramic radiographs. The presence of oral Candida was studied by fungal culture method. Bacterial samples from deepened pockets (≥4mm) were analyzed with checkerboard DNA-DNA hybridization method. VPI (15.3%) and BOP (11.2%) of the patients were modest reflecting relatively adequate oral self-care. Still 89% of the patients had at least one PPD of ≥4mm; 78.5% of the PPDs ≥6mm were found in molars. Patients had lost one third of the molars due to periodontitis and/or tooth decay. Half of the patients (53%) were Candida carriers. Bacterial analysis of subgingival plaque samples revealed bacterial species common to chronic periodontitis. AGel amyloidosis may increase the risk for periodontitis even when the oral self-care is adequate. Molar teeth appear to be mostly affected, leading to tooth loss. AGel amyloidosis as a systemic disease is related with a vast variety of symptoms with variable severity. Even though a causal relationship of the systemic disease and periodontitis has not yet been proven, increased risk for periodontal problems should be considered when examining AGel amyloidosis patients.

The Association Between Detectable Plasmatic Human Immunodeficiency Virus (HIV) Viral Load and Different Subgingival Microorganisms in Brazilian Adults With HIV: A Multilevel Analysis

This study investigates the association between detectable plasmatic human immunodeficiency virus (HIV) viral load (HVL) and high levels of periodontal- and non-periodontal-related microorganisms in the subgingival microbiota of individuals with HIV. Thirty-seven individuals with HIV were divided into two groups: 1) detectable HVL (n = 15); and 2) undetectable HVL (n = 22). Subgingival biofilm samples were obtained, and the levels of 35 microbial species were determined by the checkerboard DNA-DNA hybridization method. Periodontal clinical measures and laboratory and sociodemographic data were also registered. χ(2) test, Fisher exact test, and Mann-Whitney U test were used to compare groups. Multilevel ordinal regression models were used to test the association between HVL and the levels of 35 microbial species in subgingival biofilm, adjusted for confounders. Of the 35 species studied, 11 (31.4%) showed higher mean levels in the detectable HVL group than undetectable HVL group (P <0.001). These species included Actinomyces naeslundii II, Actinomyces israelii, Actinomyces odontolyticus, Veillonella parvula, Capnocytophaga gingivalis, Eikenella corrodens, Campylobacter concisus, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Candida albicans. Significant associations between detectable HVL and high levels of microorganisms, adjusted for confounders, were observed for A. naeslundii I, Actinomyces gerencseriae, C. gingivalis, E. corrodens, C. concisus, Prevotella nigrescens, T. forsythia, and Dialister pneumosintes. Detectable plasmatic HVL in individuals with HIV was associated with elevated levels of known periodontal pathogens, such as P. nigrescens, T. forsythia, and E. corrodens, as well as C. concisus, C. gingivalis, and D. pneumosintes in the subgingival biofilm.

Systemic antibiotics and debridement of peri‐implant mucositis. A randomized clinical trial

This RCT compared non-surgical treatment of peri-implant mucositis with or without systemic antibiotics. Forty-eight subjects received non-surgical debridement with or without systemic Azithromax (®) (4 days), and were followed during 6 months. The checkerboard DNA-DNA hybridization method was used to analyse the microbiological material. Five subjects were excluded due to antibiotic medication during follow-up. At baseline,1 and 3 months no group differences were found. Statistical analysis failed to demonstrate differences in probing pocket depths (PPD) values at 6 months (Mean diff PPD: 0.5 mm, SE: ±0.4 mm, 95% CI: -0.2, 1.3, p = 0.16). Mean% implant bleeding decreased between baseline and month 6 from 82.6% to 27.3% in the test, and from 80.0% to 47.5% in the control group (p < 0.02). Throughout the study, no study group differences in bacterial counts were found. No short-term differences were found between study groups. The clinical improvements observed at 6 months may be attributed to improvements in oral hygiene. The present study does not provide evidence for the use of systemic antibiotics in treatment of peri-implant mucositis.

Oral microbiota in Swiss adolescents

The purpose of the study was to determine the prevalence of different oral microbes in gingival plaque samples and in samples from the dorsum of the tongue in a Swiss adolescent population. Ninety-nine adolescents between 15 and 18years were enrolled. Plaque index, bleeding on probing (BOP), the periodontal screening index, and decayed missed filled tooth (DMFT) index were recorded. Samples from subgingival plaque and swabs from the tongue were analyzed by the Checkerboard DNA-DNA hybridization method. Additionally, counts of Streptococus mutans and Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were determined by real-time PCR. Periodontitis was not diagnosed in any of the subjects but all of them presented signs of gingival inflammation displaying a mean BOP of 28%. Ten (10.1%) subjects were tested positive for P. gingivalis, each 22 (22.2%) for A. actinomycetemcomitans and T. forsythia, (47.5%) for T. denticola. T. denticola and S. mutans showed a high affinity to the gingival plaque, whereas T. forsythia was often detected from the dorsum of the tongue. DMFT was associated with S. mutans counts, and BOP correlated with counts of P. gingivalis and T. denticola. The present data indicate that: (a) gingivitis but not periodontitis is a common finding among Swiss adolescents, and (b) bacteria associated with periodontitis were frequently detected in the subgingival dental plaque and on the dorsum of the tongue in Swiss adolescents with gingivitis. Although gingivitis was a frequent finding in Swiss adolescents, periodontitis was not detected in this population. The dorsum of the tongue appears to represent an important reservoir for periodontopathic bacteria.

Microbiologic Results After Non‐Surgical Erbium‐Doped:Yttrium, Aluminum, and Garnet Laser or Air‐Abrasive Treatment of Peri‐Implantitis: A Randomized Clinical Trial

The purpose of this study is to assess clinical and microbiologic effects of the non-surgical treatment of peri-implantitis lesions using either an erbium-doped:yttrium, aluminum, and garnet (Er:YAG) laser or an air-abrasive subgingival polishing method. In a 6-month clinical trial, 42 patients with peri-implantitis were treated at one time with an Er:YAG laser or an air-abrasive device. Routine clinical methods were used to monitor clinical conditions. Baseline and 6-month intraoral radiographs were assessed with a software program. The checkerboard DNA-DNA hybridization method was used to assess 74 bacterial species from the site with the deepest probing depth (PD) at the implant. Non-parametric tests were applied to microbiology data. PD reductions (mean ± SD) were 0.9 ± 0.8 mm and 0.8 ± 0.5 mm in the laser and air-abrasive groups, respectively (not significant). No baseline differences in bacterial counts between groups were found. In the air-abrasive group, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus anaerobius were found at lower counts at 1 month after therapy (P <0.001) and with lower counts in the laser group for Fusobacterium nucleatum naviforme (P = 0.002), and Fusobacterium nucleatum nucleatum (P = 0.002). Both treatments failed to reduce bacterial counts at 6 months. Porphyromonas gingivalis counts were higher in cases with progressive peri-implantitis (P <0.001). At 1 month, P. aeruginosa, S. aureus, and S. anaerobius were reduced in the air-abrasive group, and Fusobacterium spp. were reduced in the laser group. Six-month data demonstrated that both methods failed to reduce bacterial counts. Clinical improvements were limited.

Periodontitis: A Future Risk of Acute Coronary Syndrome? A Follow‐Up Study Over 3 Years

Periodontitis has been associated with cardiovascular disease. We assess if the recurrence of acute coronary syndrome (ACS) could be predicted by preceding medical and periodontal conditions. A total of 165 consecutive subjects with ACS and 159 medically healthy, matched control subjects were examined and followed for 3 years. Periodontitis was defined by alveolar bone loss. Subgingival microbial samples were studied by the checkerboard DNA-DNA hybridization method. The recurrence of ACS was found in 66 of 165 (40.0%) subjects, and a first ACS event was found in seven of 159 (4.4%) subjects among baseline control subjects. Subjects who later had a second ACS event were older (P <0.001). Significantly higher serum levels of high-density lipoprotein (P <0.05), creatinine (P <0.01), and white blood cell (WBC) counts (P <0.001) were found in subjects with future ACS. Periodontitis was associated with a first event of ACS (crude odds ratio [OR]: 10.3:1; 95% confidence interval [CI]: 6.1 to 17.4; P <0.001) and the recurrence of ACS (crude OR: 3.6:1; 95% CI: 2.0 to 6.6; P <0.001). General linear modeling multivariate analysis, controlling for age and the prediction of a future ACS event, identified that WBC counts (F = 20.6; P <0.001), periodontitis (F = 17.6; P <0.001), and serum creatinine counts (F = 4.5; P <0.05) were explanatory of a future ACS event. The results of this study indicate that recurrent ACS events are predicted by serum WBC counts, serum creatinine levels, and a diagnosis of periodontitis. Significantly higher counts of putative pathogens are found in subjects with ACS, but these counts do not predict future ACS events.

Effects of ultrasonic, electric, and manual toothbrushes on subgingival plaque composition in orthodontically banded molars

Orthodontic appliances hinder mechanical plaque control. In this study, we evaluated the effect of self-performed supragingival plaque removal with ultrasonic, electric, and manual toothbrushes on subgingival plaque composition in orthodontically banded molars. Twenty-one patients wearing fixed orthodontic appliances were assigned to this single-blind crossover study. Samples of subgingival plaque were collected from banded molars, before and after each toothbrush usage period, for quantification of 22 bacterial species by the checkerboard DNA-DNA hybridization method. For each crossover, patients used a toothbrush for 30 days, followed by a washout period of 14 days. The prevalence of Tannerella forsythia decreased significantly after a month of electric brush usage. In the manual brush group, the prevalences of Selenomonas noxia, Streptococcus sanguinis, and Prevotella melaninogenica also decreased significantly. However, there were no significant differences in the prevalences and levels of bacteria after usage of the ultrasonic brush. Intergroup comparisons showed no statistical differences among the 3 brushes for the microbiologic parameters. All 3 brushes generally reduced bacterial prevalences, and, although electric and manual toothbrushes showed some isolated significant variations, we found no superiority with any toothbrush type when used three times daily for 2 minutes on microbiologic parameters in orthodontically banded molars.

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

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. 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. 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). 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.

Microbiological Findings in Subjects With Asymptomatic Oral Lichen Planus: A Cross‐Sectional Comparative Study

The bacterial colonization of the oral mucosa was evaluated in patients with asymptomatic oral lichen planus (OLP) and compared to the microbiologic status in mucosally healthy subjects. Bacteria from patients with clinically and histopathologically diagnosed OLP from the Stomatology Service, Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern, were collected with a non-invasive swab system. Samples were taken from OLP lesions on the gingiva and from non-affected sites on the contralateral side of the mouth. The control population did not have OLP and was recruited from the student clinic. All samples were processed with the checkerboard DNA-DNA hybridization method using well-defined bacterial species for the analysis. Significantly higher bacterial counts of Bacteroides ureolyticus (P = 0.001), Dialister species (sp.) (P = 0.006), Staphylococcus haemolyticus (P = 0.007), and Streptococcus agalactiae (P = 0.006) were found in samples taken from OLP lesions compared to sites with no clinical evidence of OLP. Significantly higher bacterial counts were found for Capnocytophaga sputigena, Eikenella corrodens, Lactobacillus crispatus, Mobiluncus curtisii, Neisseria mucosa, Prevotella bivia, Prevotella intermedia, and S. agalactiae at sites with lesions in subjects with OLP compared to sites in control subjects (P <0.001). Microbiologic differences were found between sites with OLP and sites in subjects without a diagnosis of OLP. Specifically, higher counts of staphylococci and S. agalactiae were found in OLP lesions.

Similarities in the Subgingival Microbiota Assessed by a Curet Sampling Method at Sites With Chronic Periodontitis

The goal of this study was to determine whether site-specific differences in the subgingival microbiota could be detected by the checkerboard method in subjects with periodontitis. Subjects with at least six periodontal pockets with a probing depth (PD) between 5 and 7 mm were enrolled in the study. Subgingival plaque samples were collected with sterile curets by a single-stroke procedure at six selected periodontal sites from 161 subjects (966 subgingival sites). Subgingival bacterial samples were assayed with the checkerboard DNA-DNA hybridization method identifying 37 species. Probing depths of 5, 6, and 7 mm were found at 50% (n = 483), 34% (n = 328), and 16% (n = 155) of sites, respectively. Statistical analysis failed to demonstrate differences in the sum of bacterial counts by tooth type (P = 0.18) or specific location of the sample (P = 0.78). With the exceptions of Campylobacter gracilis (P <0.001) and Actinomyces naeslundii (P <0.001), analysis by general linear model multivariate regression failed to identify subject or sample location factors as explanatory to microbiologic results. A trend of difference in bacterial load by tooth type was found for Prevotella nigrescens (P <0.01). At a cutoff level of > or = 1.0 x 10(5), Porphyromonas gingivalis and Tannerella forsythia (previously T. forsythensis) were present at 48.0% to 56.3% and 46.0% to 51.2% of sampled sites, respectively. Given the similarities in the clinical evidence of periodontitis, the presence and levels of 37 species commonly studied in periodontitis are similar, with no differences between molar, premolar, and incisor/cuspid subgingival sites. This may facilitate microbiologic sampling strategies in subjects during periodontal therapy.

A Microbiological Profile of Symptomatic Teeth with Primary Endodontic Infections

The aim of this study was to evaluate the composition of the microbiota of primary endodontic infections associated with symptomatic teeth. Samples were collected by means of a #15 H-type file and 2 sterile paper points from 60 symptomatic (n = 30) or asymptomatic (n = 30) single-rooted teeth with necrotic pulp. The presence of 40 bacterial species was determined by the checkerboard DNA-DNA hybridization method. The species found in higher counts (×105) in symptomatic cases were Fusobacterium nucleatum ssp. vincentii, Veillonella parvula, Treponema socranskii, Enterococcus faecalis, and Campylobacter gracilis and in asymptomatic cases were F. nucleatum ssp. vincentii, Fusobacterium nucleatum ssp. nucleatum, E. faecalis, Eubacterium saburreum, and Neisseria mucosa. Total bacterial counts and counts of Tannerella forsythia were significant higher in symptomatic cases (p < 0.05), whereas levels of Propionibacterium acnes were reduced in this group of teeth. The data of the present investigation suggested an association between higher total bacterial counts and levels of T. forsythia and the presence of pain.