Phenotypes, serotypes and antibiotic susceptibility of Swedish Porphyromonas gingivalis isolates from periodontitis and periodontal abscesses

This study was conducted to reveal phenotypic, serological subtypes and antibiotic susceptibility among fresh isolates of Porphyromonas gingivalis in a Swedish population with periodontitis and periodontal abscess. Fifty-five subgingival strains were isolated and tentatively designated as P. gingivalis from 55 consecutive paper-point samples taken from 51 patients with periodontitis (at least one site with >6-mm pocket depth) in Sweden and were sent in for microbiological evaluation. Eight P. gingivalis strains from periodontal abscesses were also included. Four P. gingivalis strains served as reference and another four type strains were included. The strains were characterized by colony morphology, biochemical tests, enzyme profile, gas-liquid chromatography and antibiotic susceptibility. The strains were further characterized for whole cell protein profiles using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and were identified to serotype by specific monoclonal antibodies. Among the 55 P. gingivalis strains 35 had smooth (S), 13 rough (R) and seven semi-rough colony morphologies. All strains were phenotypically homogeneous in biochemical tests, enzyme profile and antibiotic susceptibility. All strains produced phenylacetic acid and alpha-fucosidase. Almost all (96%) of the subgingival strains, but relatively fewer (62%) of the abscess strains, belonged to serotype A. Two subgingival and three abscess strains were classified as serotype B. No specific SDS-PAGE protein profiles were recorded for the two serotypes. The P. gingivalis strains from Swedish periodontitis cases showed homogeneity in terms of biochemical phenotypes and antibiotic susceptibility patterns. The strains fell into two serotypes, of which serotype A predominated in the periodontitis cases and serotype B was overrepresented in periodontal abscesses.     

Response to alkaline stress by root canal bacteria in biofilms

AIM: To determine whether bacteria isolated from infected root canals survive alkaline shifts better in biofilms than in planktonic cultures. METHODOLOGY: Clinical isolates of Enterococcus faecalis, Lactobacillus paracasei, Olsenella uli, Streptococcus anginosus, S. gordonii, S. oralis and Fusobacterium nucleatum in biofilm and planktonic cultures were stressed at pH 10.5 for 4 h, and cell viability determined using the fluorescent staining LIVE/DEAD BacLight bacterial viability kit. In addition, proteins released into extracellular culture fluids were identified by Western blotting. RESULTS: Enterococcus faecalis, L. paracasei, O. uli and S. gordonii survived in high numbers in both planktonic cultures and in biofilms after alkaline challenge. S. anginosus, S. oralis and F. nucleatum showed increased viability in biofilms compared with planktonic cultures. Alkaline exposure caused all planktonic cultures to aggregate into clusters and resulted in a greater extrusion of cellular proteins compared with cells in biofilms. Increased levels of DnaK, HPr and fructose-1,6-bisphosphate aldolase were observed in culture fluids, especially amongst streptococci. CONCLUSIONS: In general, bacteria isolated from infected roots canals resisted alkaline stress better in biofilms than in planktonic cultures, however, planktonic cells appeared to use aggregation and the extracellular transport of specific proteins as survival mechanisms.     

The effects of amalgam restorations on plasma mercury levels and total antioxidant activity

Objective

This study evaluated the effects of amalgam restorations on plasma mercury levels and total antioxidant activities (TAA).

Design

The study was comprised of 48 subjects ranging in age from 20 to 32 years. Of these, 33 had dental amalgam restorations and 15 had no dental amalgam restorations. In those patients with amalgams, the total number of amalgam restorations and surfaces were counted, and the total and occlusal areas (mm²) of restorations were measured using a Counting Measurement Machine. Blood samples were collected from all participants. Plasma mercury levels were measured using an Atomic Absorption Spectrometer and Hydride System, and plasma TAA levels were measured using an Antioxidant Assay Kit. Statistical analysis was performed using the SPSS 10.01 software program. Data was evaluated by t test and correlation analysis.

Results

Plasma mercury (P-Hg) levels were found to be significantly higher in subjects with amalgam restorations when compared to subjects without amalgams (p < 0.01); the differences in P-TAA levels between subjects with and without amalgams were not found to be statistically significant (p > 0.05). No significant correlations were found between P-Hg concentrations and P-TAA levels (p > 0.05). Significant positive correlations were found between P-Hg concentrations and the number of amalgam restorations (p < 0.01), number of amalgam surfaces (p < 0.05), total amalgam surface area (p < 0.05) and amalgam occlusal surface area (p < 0.01). However, no significant correlations were found between these parameters and P-TAA (p > 0.05).

Conclusions

The results of our study showed that dental amalgams are a major source of plasma mercury; however, amalgam restorations were not found to have a significant effect on plasma-total antioxidant activities.     

Gram-positive rods prevailing in teeth with apical periodontitis undergoing root canal treatment

AIMS: To identify Gram-positive rods from root canals of teeth with apical periodontitis and to examine their associations with other species. METHODOLOGY: Consecutive root canal samples (RCSs) from 139 teeth undergoing root canal treatment were analyzed prospectively for cultivable microbes. Gram-positive rods in the first RCS submitted after chemo-mechanical preparation were categorised to genus level by selective media and gas-liquid chromatography (GLC), and identified to species level by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Associations between organisms were measured by odds ratios (OR). RESULTS: In the first samples submitted a total of 158 Gram-positive rods, 115 Gram-positive cocci, 26 Gram-negative rods and 9 Gram-negative cocci, were identified. At genus levels Gram-positive rods were classified into: Lactobacillus spp. (38%), Olsenella spp. (18%), Propionibacterium spp. (13%), Actinomyces spp. (12%), Bifidobacterium spp. (13%) and Eubacterium spp. (6%). The most frequent species were Olsenella uli, Lactobacillus paracasei and Propionibacterium propionicum. In subsequent samples taken during treatment, Gram-positive rods were also identified, although the number of strains was considerably reduced. Positive associations were observed between members of the genus lactobacilli and Gram-positive cocci (OR>2). CONCLUSIONS: Olsenella uli and Lactobacillus spp. predominated over other Gram-positive rods. A possible association exists between Lactobacillus spp. and Gram-positive cocci in root canals of teeth with apical periodontitis receiving treatment.     

Real-time quantitative polymerase chain reaction and culture analyses of Enterococcus faecalis in root canals

Reports on the prevalence of Enterococcus faecalis in root canals vary considerably, potentially because of variations in clinical sampling and sample analysis methods. This study compared culture and real-time quantitative polymerase chain reaction (qPCR) to detect and quantify E. faecalis in the same root canal sample. Consecutive root canal samples obtained from primary infection (n = 40) and retreatment (n = 48) cases were divided into two equal aliquots that were independently analyzed using culture and qPCR by investigators blinded to the analysis results of the other sample. E. faecalis was detected in 10.2% and 79.5% of samples by culture and qPCR, respectively (p < 0.0001; McNemar's test). E. faecalis was detected in more retreatment than primary infection samples (89.6% versus 67.5%; p = 0.01, Fisher's exact test). qPCR reported a significantly higher prevalence of E. faecalis in endodontic samples than culture techniques.     

Inactivation of local root canal medicaments by dentine: an in vitro study

AIMS: The aim of the study was to investigate the inactivation by dentine of the antibacterial activity of various commonly used local root canal medicaments. METHODOLOGY: The medicaments tested were saturated calcium hydroxide solution, 1% sodium hypochlorite, 0.5% and 0.05% chlorhexidine acetate, and 2/4% and 0.2/0.4% iodine potassium iodide. Dentine was sterilized by autoclaving and crushed into powder with a particle size of 0.2-20 microns. Aliquots of dentine suspension were incubated with the medicaments in sealed test tubes at 37 degrees C for 24 h or 1 h before adding the bacteria. In some experiments bacteria were added simultaneously with dentine powder and the medicament. Enterococcus faecalis A197A was used as a test organism. Samples for bacterial culturing were taken from the suspensions at 5 min, 1 h and 24 h after adding the bacteria. RESULTS: Dentine powder had an inhibitory effect on all medicaments tested. The effect was dependent on the concentration of the medicament as well as on the length of the time the medicament was preincubated with dentine powder before adding the bacteria. The effect of calcium hydroxide on E. faecalis was totally abolished by the presence of dentine powder. Similarly, 0.2/0.4% iodine potassium iodide lost its effect after preincubation for 1 h with dentine before adding the bacteria. The effect of 0.05% chlorhexidine and 1% sodium hypochlorite on E. faecalis was reduced but not totally eliminated by the presence of dentine. No inhibition could be measured when full strength solutions of chlorhexidine and iodine potassium iodide were used in killing E. faecalis. CONCLUSIONS: The dentine powder model appears to be an efficient tool for the study of interactions between local endodontic medicaments, dentine, and microbes.     

Some aspects of the orthodontic-prosthetic approach for adult patients. A case report

Authors present a case report of an adult patient treated with the combination of orthodontic and prosthetic treatment methods. The importance of periodontal and psychological management of such patients is pointed out.     

Effects of phosphoric acid on bovine enamel bleached with carbamide peroxide

The aim of this study was to measure the demineralization capacity of 37% phosphoric acid on bovine enamel at different time-points after bleaching with 30% carbamide peroxide. Five, 4 × 4-mm sections were obtained from the enamel of 10 bovine incisors. After applying 30% carbamide peroxide (Vivastyle) for 90 min, specimens were stored in artificial saliva for 0, 24, 72 h, or 7 d and then immersed in 37% phosphoric solution. At 15, 30, 60, 90, and 120 s, 5-ml aliquots were extracted. A control group of specimens was not bleached. Ca²⁺ concentrations were measured by atomic absorption spectrophotometry. A larger amount of Ca²⁺ was extracted from enamel by phosphoric acid after the application of 30% carbamide peroxide. Twenty-four hours after bleaching, significantly more Ca²⁺ was extracted from bleached than from control specimens at all time-points, and this greater susceptibility to the action of the acid persisted for at least 1 wk after bleaching.