Biomedical subjects

C M Sedgley

Publications and source records attributed to C M Sedgley.

16 recordsLinked to original sources

PubMed2005

Influence of irrigant needle depth in removing bioluminescent bacteria inoculated into instrumented root canals using real-time imaging in vitro.

AIM: To test the hypothesis that the mechanical efficacy of irrigation in reducing bacteria in the root canal is dependent on depth of placement of the irrigation needle. METHODOLOGY: The root canals of 30 permanent cuspids were instrumented to apical size 60 using a crown-down technique. A suspension of the bioluminescent reporter strain Pseudomonas fluorescens 5RL was inoculated into each canal of sterilized teeth. Emission of bioluminescence (photons s(-1)) from each tooth was quantified on four sequential occasions using luminometry and bioluminescence imaging: (i) background, (ii) after inoculation, (iii) after irrigating the inoculated teeth with 3 mL of a nonantimicrobial irrigant delivered either 1 mm (group 1, n = 15) or 5 mm (group 2, n = 15) from working length (WL) using a 28G safety-ended irrigating needle, (iv) after an additional 3 mL irrigation (total 6 mL). Intragroup and intergroup comparisons were made using Wilcoxon matched pairs and Mann-Whitney tests, respectively. RESULTS: In group 1, there was a mean log10 decrease in bacteria of 0.68 +/- 0.26 after 3 mL of irrigant compared with 1.19 +/- 0.48 after 6 mL (P < 0.001); in group 2 the mean log10 decrease was 0.58 +/- 0.28 after 3 mL of irrigant compared with 0.69 +/- 0.35 after 6 mL (P < 0.02) (Wilcoxon matched pairs). Using 3 mL of irrigant, needle depth did not have a significant effect on reduction of intracanal bacteria (P = 0.407), but the effect became significant when 6 mL of irrigant was used (P < 0.002) (Mann-Whitney tests). CONCLUSIONS: The mechanical efficacy of 6 mL of irrigant in reducing intracanal bacteria was significantly greater when delivered 1 mm compared with 5 mm from WL.

Colony Count, Microbial↗

PubMed2005

Survival of Enterococcus faecalis in root canals ex vivo.

AIM: The hypotheses tested in this study were that: (i) Enterococcus faecalis can survive long-term entombment in root filled teeth without additional nutrients, (ii) initial cell density influences the survival of E. faecalis in instrumented root canals and (iii) gelatinase-production capacity influences the survival of E. faecalis in root canals. METHODOLOGY: The root canals of 150 extracted single canal teeth were instrumented to apical size 60 and divided into six groups of 25. Within each group 10 canals were inoculated with either gelatinase-producing E. faecalis OG1-S and the other 10 with its gelatinase-defective mutant E. faecalis OG1-X. Five canals per group were kept as uninoculated controls. The root canals in groups 1 and 2 were inoculated with 10(6) bacteria, incubated for 48 h at 37 degrees C then filled with gutta-percha and zinc-oxide eugenol sealer. Root canals were inoculated with 10(6), 10(5), 10(4) and 10(3) bacteria in groups 3-6, respectively, and left unfilled. All teeth were sealed coronally with glass-ionomer cement. After 6- (groups 1, 3-6) and 12-month (group 2) incubation at 37 degrees C in 100% humidity, root fragments were analysed for presence of E. faecalis, using culture, polymerase chain reaction and histological methods. RESULTS: Viable E. faecalis was recovered from all root filled teeth and from 95-100% of unfilled inoculated teeth. Initial cell density and gelatinase production did not influence the recovery of viable E. faecalis (P > 0.05; chi-square test). Enterococcus faecalis 16S rRNA gene products were present in all inoculated teeth and absent in all noninoculated controls. Dentinal tubule infection was evident under light microscopy in sections from inoculated teeth after 48-h, 6- and 12-month incubation. CONCLUSIONS: Enterococcus faecalis inoculated into root canals maintained viability for 12-months ex vivo. The clinical implications are that viable E. faecalis entombed at the time of root filling could provide a long-term nidus for subsequent infection.

Adaptation, Physiological↗

PubMed2005

Virulence, phenotype and genotype characteristics of endodontic Enterococcus spp.

BACKGROUND/AIMS: Enterococci have been implicated in persistent root canal infections but their role in the infection process remains unclear. This study investigated the virulence, phenotype and genotype of 33 endodontic enterococcal isolates. METHODS: Phenotypic tests were conducted for antibiotic resistance, clumping response to pheromone, and production of gelatinase, hemolysin and bacteriocin. Genotype analysis involved polymerase chain reaction amplification of virulence determinants encoding aggregation substances asa and asa373, cytolysin activator cylA, gelatinase gelE, gelatinase-negative phenotype ef1841/fsrC, adherence factors esp and ace, and endocarditis antigen efaA. Physical DNA characterization involved pulsed-field gel electrophoresis of genomic DNA, and plasmid analysis. RESULTS: Potential virulence traits expressed included production of gelatinase by Enterococcus faecalis (n=23), and response to pheromones in E. faecalis culture filtrate (n=16). Fourteen strains produced bacteriocin. Five strains were resistant to tetracycline and one to gentamicin, whereas all were susceptible to ampicillin, benzylpenicillin, chloramphenicol, erythromycin, fusidic acid, kanamycin, rifampin, streptomycin and vancomycin. Polymerase chain reaction products encoding efaA, ace, and asa were detected in all isolates; esp was detected in 20 isolates, cylA in six isolates, but asa373 was never detected. The gelatinase gene (gelE) was detected in all isolates of E. faecalis (n=31) but not in Enterococcus faecium (n=2); a 23.9 kb deletion sequence corresponding to the gelatinase-negative phenotype was detected in six of the eight E. faecalis isolates that did not produce gelatinase. Pulsed-field gel electrophoresis and plasmid analyses revealed genetic polymorphism with clonal types evident. Plasmid DNA was detected in 25 strains, with up to four plasmids per strain and a similar (5.1 kb) plasmid occurring in 16 isolates. CONCLUSIONS: Phenotypic and genotypic evidence of potential virulence factors were identified in endodontic Enterococcus spp., specifically production of gelatinase and response to pheromones.

Adult↗

PubMed2004

Quantitative real-time PCR detection of oral Enterococcus faecalis in humans.

OBJECTIVE: Enterococcus faecalis is consistently associated with recurrent root canal infections. Only low concentrations of E. faecalis in the human mouth have been demonstrated by culture techniques. Quantitative detection strategies more sensitive than culturing, such as quantitative PCR (qPCR), could provide more illuminating data. DESIGN: Thirty outpatients attending the University of Michigan Graduate Endodontic Clinic for endodontic treatment provided oral rinse samples that were analysed for E. faecalis using qPCR and microbiological culturing. A SYBR Green I qPCR protocol was developed for the quantifiable detection of E. faecalis and total bacteria in oral rinse samples using primers designed to target the 16S rRNA gene. Annealing temperature and primer, magnesium ion, and dimethyl sulfoxide concentrations were investigated for optimisation of the protocol; a minimum sensitivity limit of 23 rRNA copies (an estimated six E. faecalis cells) was established for E. faecalis in pure culture, and 104 rRNA copies (an estimated 26 E. faecalis cells) in mixed culture. RESULTS: In qPCR assays, based on extrapolation from estimated rRNA gene copy numbers, E. faecalis comprised 0.0006-0.0047% of a total bacteria load that ranged from 5.92 x 10(5) to 5.69 x 10(7) cells/ml of oral rinse. E. faecalis was detected in five (17%) samples in concentrations from 114 to 490 cells/ml. In parallel culture assays E. faecalis were detected in only two samples (7%) of the five identified by qPCR and in concentrations 30 and 240 CFU/ml. CONCLUSIONS: qPCR reported a higher incidence of E. faecalis in oral rinse samples than culture techniques and afforded greater sensitivity.

Colony-Forming Units Assay↗

PubMed2004

Prevalence, phenotype and genotype of oral enterococci.

This study investigated the prevalence, phenotype and genotype of oral enterococci. Enterococci were detected in oral rinse samples from 11% of 100 patients receiving endodontic treatment and 1% of 100 dental students with no history of endodontic treatment (P=0.0027). All enterococcal isolates were identified as Enterococcus faecalis. Viable counts ranged from 1 x 10 to 6 x 103 colony forming units per mL of oral rinse sample. Potential virulence traits expressed by oral E. faecalis strains included production of hemolysin (n=4) and gelatinase (n=4), and response to pheromones in E. faecalis culture filtrate (n=1). Six strains produced bacteriocin. All strains were susceptible to ampicillin, benzylpenicillin, gentamicin and vancomycin. There was no evidence of metal-ion resistance. One isolate produced hemolysin, gelatinase and bacteriocin, was resistant to several antibiotics, and responded to the pheromone cPD1. Pulsed-field gel electrophoresis and plasmid analysis showed that oral E. faecalis exhibited widespread genetic polymorphism, with plasmids detected in seven strains.

Adult↗

PubMed2003

The antimicrobial efficacy of 'MGP' gutta-percha in vitro.

AIM: To determine whether 'MGP' gutta-percha (Westport, CT, USA), a commercially available gutta-percha containing iodoform, inhibits the growth of potential endodontic pathogens. METHODOLOGY: Inocula of Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Streptococcus sanguis, Fusobacterium nucleatum and Actinomyces odontolyticus were spread onto the surface of agar plates. 'MGP' gutta-percha cones presoaked in sterile water were transferred to the inoculated agar and incubated at 37 degrees C aerobically or anaerobically as required for optimal growth. Identical studies were performed using iodoform-free gutta-percha and sterile paper disks saturated with 10% povidone-iodine. Following incubation, zones of inhibition around the 'MGP' gutta-percha, iodoform-free gutta-percha and disks were evaluated. RESULTS: Povidone-iodine inhibited all the strains. Iodoform-free gutta-percha inhibited S. sanguis and A. odontolyticus. 'MGP' gutta-percha inhibited S. aureus, S. sanguis, A. odontolyticus and F. nucleatum. Neither iodoform-free gutta-percha nor 'MGP' gutta-percha inhibited growth of E. faecalis, E. coli or P. aeruginosa. CONCLUSIONS: Compared to iodoform-free gutta-percha, iodoform-containing 'MGP' gutta-percha had an inhibitory effect in vitro on S. aureus and F. nucleatum, but not on E. faecalis, E. coli or P. aeruginosa.

Actinomyces↗

PubMed2003

Orthograde retreatment and apexification after unsuccessful endodontic treatment, retreatment and apicectomy.

AIM: To describe a case where a second orthograde retreatment was successful in the management of an infected mandibular right first molar that previously had received both orthograde and retrograde treatments. SUMMARY: Periapical surgery is unlikely to be successful unless the root canal system has been adequately debrided and sealed. A case is described where orthograde endodontic treatment, retreatment and apicectomy were unsuccessful in the management of and infected mandibular right first molar. The periapical radiolucency eventually disappeared following a second orthograde retreatment. Teh second retreatment included 12 months of intracanal calcium hydroxide placement to promote apexification, thus allowing subsequent controlled obturation with gutta percha and AH26. At a 5-year review following completion of treatment, the tooth remained asymptomatic and was in normal function. KEY LEARNING POINTS: Orthograde retreatment is a treatment option to manage refractory lesions in teeth that have previously received endodontic treatment, retreatment and apicectomy. Orthograde retreatment using long-term intracanal calcium hydroxide can help promote root-end closure of a resected apex.

Apicoectomy↗

PubMed1998

Antimicrobial susceptibility of oral isolates of Enterobacter cloacae and Klebsiella pneumoniae from a southern Chinese population.

The antibiotic susceptibilities of 59 Enterobacter cloacae and 39 Klebsiella pneumoniae human oral isolates collected from a southern Chinese population in Hong Kong were investigated for their susceptibility to eight antibiotics: ampicillin, cephalothin, cefuroxime, ceftazidime, ciprofloxacin, gentamicin, tetracycline and trimethoprim/sulfamethoxazole using the E-Test method for direct quantification of minimum inhibitory concentrations. Most strains were sensitive to all antibiotics except ampicillin and cephalothin. Ampicillin resistance was exhibited by 82% of K. pneumoniae and 69% of E. cloacae isolates. Eighty-eight percent of E. cloacae isolates were resistant to cephalothin. Several strains fell within the intermediate category of sensitivity for ampicillin (E. cloacae and K. pneumoniae), cefuroxime (E. cloacae) and tetracycline (K. pneumoniae). Comparison with other Hong Kong data suggests that resistance rates to cefuroxime, ceftazidime, ciprofloxacin, gentamicin, tetracycline and trimethoprim/sulfamethoxazole exhibited by the oral isolates are generally lower than in enterobacters and Klebsiella spp. isolated from urine, skin and soft tissues in Hong Kong populations.

Adult↗

PubMed1997

A 4-year longitudinal study of the oral prevalence of enteric gram-negative rods and yeasts in Chinese children.

A 4-year longitudinal study of the oral prevalence of enteric gram-negative rods and yeasts in 116 Chinese primary school children in Hong Kong was conducted. The oral prevalence of enteric gram-negative rods for each consecutive year was 25.3%, 37.0%, 24.0% and 25.8% respectively, with a weighted mean of 27.9%. Enterobacteriaceae, which comprised 57% of all enteric gram-negative rods, were more common in children with no caries experience. The oral prevalence of yeasts for each consecutive year was 7.7%, 12.0%, 14.4% and 15.5% respectively, with a weighted mean of 12.5%. Candida albicans comprised 84% of all yeasts isolated. Oral yeast carriage was significantly associated with caries prevalence. While the oral prevalence of enteric gram-negative rods in primary school children in Hong Kong may be higher than in other parts of the world, repeated isolation of either enteric gram-negative rods or Candida spp. from individual children over the 4-year study period was rare, suggesting that carriage of these organisms is transient.

Candida↗

PubMed1996

The oral prevalence of aerobic and facultatively anaerobic gram-negative rods and yeasts in semi-recluse human vegetarians.

Limited data exist on the oral ecology of vegetarians. Hence the dental and periodontal status, and the oral prevalence of aerobic and facultatively anaerobic Gram-negative rods (AGNR) and yeasts, were studied in 36 semi-recluse, vegetarian, Buddhist monks and nuns in Hong Kong. The oral prevalence of AGNR and yeasts was 61.1% and 33.3%. There was no correlation between the prevalence of AGNR and/or yeasts and the incidence of carious or filled teeth and the health status of the periodontium. Rather, the results of this study combined with those of previous studies suggest that increasing age and the consumption of food prepared in communal kitchens might be more important contributory factors in the oral prevalence of AGNR than the nature of the diet itself or the health of the dentition and periodontium.

Adult↗

PubMed1996

The adherence of oral isolates of Enterobacteriaceae to HeLa cells. An in vitro method using image analysis.

An in vitro model and an image analysis were designed to improve on existing quantification methods in the assessment of the adherence of Enterobacteriaceae to human epithelial cell monolayers. Adherence to HeLa cell monolayers of three oral isolates and one type strain from each of four species of Enterobacteriaceae over two incubation time periods was examined. Correction for actual cell area and a cube root transformation of the data to stabilize variance were applied. While behaviour varied between strains within species, E. cloacae was the most, and K. pneumoniae the least, adherent species irrespective of the incubation period. Increasing the incubation period from 30 min to 60 min resulted in greater adherence for E. cloacae, E. coli, and C. freundii, but not K. pneumoniae strains. The method permits the reliable measurement and valid analysis of the adherence of Enterobacteriaceae to cultured epithelial cell monolayers.

Cell Adhesion↗

PubMed1994

The oral prevalence of aerobic and facultatively anaerobic gram-negative rods and yeasts in Hong Kong Chinese.

Saline oral rinse samples were obtained from 300 community-dwelling Hong Kong Chinese attending an outpatient dental clinic to determine the oral prevalence of aerobic and facultatively anaerobic Gram-negative rods (AGNR) and yeasts. The oral prevalence of AGNR was 41.7%. Enterobacteriaceae species comprised 73% of all AGNR isolated, with an overall prevalence of 32%. There was no difference in prevalence between females (n = 190) and males (n = 110). Morning samples (n = 154) yielded a significantly higher prevalence of AGNR (54.5%) and Enterobacteriaceae (42.2%) than afternoon samples (n = 146) (28.1 and 21.2%, respectively; p < 0.01, p < 0.01). Subjects over 50 yr had a significantly higher prevalence of AGNR than those aged 30-49 yr (p < 0.01). The most commonly isolated AGNR species were Enterobacter cloacae and Klebsiella pneumoniae pneumoniae. The oral prevalence of yeasts was 24%, with Candida albicans forming 77% of all yeasts isolated. Subjects taking medication (n = 38) or wearing dentures (n = 38) had a significantly higher oral yeast prevalence of 36.8 (p < 0.05) and 44.7% (p < 0.01), respectively. Yeast prevalence was significantly higher in subjects over 50 yr than those aged 30-49 yr (p < 0.05) and 15-29 yr (p < 0.05). Comparisons with previous studies suggest that the oral prevalence of AGNR in Chinese may be higher in Hong Kong than in other parts of the world.

Adolescent↗

PubMed1994

Oral and oropharyngeal prevalence of Enterobacteriaceae in humans: a review.

Members of the Enterobacteriaceae family are widely distributed in nature and exhibit substantial diversity in ecology, host range and pathogenic potential for man. While wide discrepancies in methodology exist between epidemiological studies, the available data indicate an increased prevalence of oral and/or oropharyngeal Enterobacteriaceae carriage in patients with illnesses of varying severity compared with healthy subjects. This paper reviews the prevalence of Enterobacteriaceae in the oral and oropharyngeal region of healthy human subjects and those affected by different disease entities, and discusses the complexities associated with collating and interpreting such data. The effect of antimicrobials and antiseptics on oral and oropharyngeal Enterobacteriaceae has also been reviewed, while highlighting the gaps in knowledge and future research directions.

Adolescent↗

PubMed1993

Long-term retention of a paper point in the periapical tissues: a case report.

A case is reported of a 15 mm paper point found in the periapical area of a maxillary right central incisor during endodontic surgery for an unresolved periapical radiolucency. The paper point had been present for between five and nine years. Histopathological examination showed the presence of chronically inflamed fibrous connective tissue with a number of foreign body giant cells and focal collections of chronic inflammatory cells. SEM comparison of the specimen with a similarly treated fresh paper point showed no indication of degradation of the paper point by host defense cells.

Adult↗

PubMed1992

Are endodontically treated teeth more brittle?

This study compared biomechanical properties (punch shear strength, toughness, hardness, and load to fracture) of 23 endodontically treated teeth (mean time since endodontic treatment: 10.1 yr) and their contralateral vital pairs. Analyses using paired t tests revealed no significant differences in punch shear strength, toughness, and load to fracture between the two groups. Vital dentin was 3.5% harder than dentin from contralateral endodontically treated teeth (p = 0.002). The similarity between the biomechanical properties of endodontically treated teeth and their contralateral vital pairs indicates that teeth do not become more brittle following endodontic treatment. Other factors may be more critical to failure of endodontically treated teeth.

Adult↗

PubMedDate not supplied

The influence of incubation conditions on the adherence of oral Enterobacteriaceae to HeLa cells.

The adherence of 12 oral isolates and 4 type strains of Enterobacteriaceae (equally representing Enterobacter cloacae, Klebsiella pneumoniae, Escherichia coli and Citrobacter freundii) to HeLa cell monolayers following five different incubation conditions (sucrose, D-mannose, serum, MEM and Candida albicans GDH 1957) was investigated. Incubation with sucrose and D-mannose resulted in the greatest and least adherence, respectively. The presence of preadherent C. albicans GDH 1957 on the HeLa cells tended to enhance the adherence of certain strains of E. cloacae and C. freundii, but had no overall impact on Enterobacteriaceae adherence. While heterogeneity of behaviour existed between strains within species, E. cloacae was the most, and K. pneumoniae the least, adherent species irrespective of incubation conditions. Haemagglutination assays indicated the presence of mannose-resistant type 1 fimbriae associated with all Enterobacteriaceae. In clinical terms, the variations in adherence properties observed in vitro may contribute to an understanding of the different prevalence rates of oral Enterobacteriaceae reported in the literature.

Bacterial Adhesion↗