The antimicrobial effect of calcium hydroxide in root canals pretreated with 5% iodine potassium iodide

Abstract— Calcium hydroxide (CH) is often used as a routine interappointment dressing during the endodontic treatment of teeth with apical periodontitis. However, it fails to consistently produce sterile root canals. The present study was set up to find out whether an antimicrobial strategy including the use of CH could be made more effective if: 1) canals were pretreated with 5% iodine potassium iodide (IPI), and 2) the dressing period was extended up to 2 months. Fifty human teeth, with radiographically verified apical periodontitis, were microbiologically sampled. After chemomechanical preparation the canals were pretreated with IPI for 3–7 days. Teeth where microorganisms persisted were then treated with CH for 2 months. Following instrumentation and dressing with IPI, 43 bacterial strains were recovered in 22 of the teeth. Samples obtained after the CH dressing period disclosed growth of 13 facultative and two strict anaerobic strains in 10 teeth. Enterococcus faecalis was identified in two specimens. In conclusion, the present study gave no evidence for an increased antimicrobial effect of CH if it was left for longer periods in the root canal. Although pretreatment with IPI from a quantitative point of view did not seem to add antimicrobial power, it might reduce the frequency of persisting strains of E. faecalis .     

The diagnostic accuracy of microbiologic root canal sampling and the influence of antimicrobial dressings

Reit C, Molander A, Dahlén G. The diagnostic accuracy of microbiologic root canal sampling and the influence of antimicrobial dressings. Endod Dent Traumatol 1999; 15: 278–283.© Munksgaard, 1999.
Abstract — The routine approach to endodontic treatment of teeth with apical periodontitis often involves an interappointment dressing with calcium hydroxide. However, investigations have demonstrated a negative influence of calcium hydroxide on the accuracy of microbiological root canal sampling (MRS). The aim of the present study was to investigate whether the use of a fluid dressing like 5% iodine potassium iodide (IPI) would increase the accuracy of MRS. Following instrumentation of 50 teeth with radiographically verified apical periodontitis the root canals received IPI as an intra-canal dressing. One week after closure canals were sampled, "test sample" (TS), and then left filled with sampling fluid and temporarily sealed. Seven days later a "gold standard" (GS) sample was obtained. Bacteria were recovered in 22 teeth (44%) in TS as well as in GS. Fifteen teeth (30%) were positive for growth in both samples. Using the detection level "very sparse growth" of microbes the sensitivity and specificity of MRS reached 68% and 75%, respectively. In an earlier study, following the same experimental protocol, but with calcium hydroxide as intracanal dressing, the corresponding values were 33% and 81%. In 25% of these cases bacteria persisted in the canals. As compared to calcium hydroxide, the use of IPI resulted in improved test accuracy, but loss of antibacterial capacity. Conclusively, intracanal dressings seem to vary in their influence on the microbiologic test performance as well as in their antibacterial efficacy. In a clinical situation the choice of interappointment dressing should include consideration of these potentially conflicting properties.     

The diagnostic accuracy of microbiologic root canal sampling and the influence of antimicrobial dressings

The routine approach to endodontic treatment of teeth with apical periodontitis often involves an interappointment dressing with calcium hydroxide. However, investigations have demonstrated a negative influence of calcium hydroxide on the accuracy of microbiological root canal sampling (MRS). The aim of the present study was to investigate whether the use of a fluid dressing like 5% iodine potassium iodide (IPI) would increase the accuracy of MRS. Following instrumentation of 50 teeth with radiographically verified apical periodontitis the root canals received IPI as an intracanal dressing. One week after closure canals were sampled, "test sample" (TS), and then left filled with sampling fluid and temporarily scaled. Seven days later a "gold standard" (GS) sample was obtained. Bacteria were recovered in 22 teeth (44%) in TS as well as in GS. Fifteen teeth (30%) were positive for growth in both samples. Using the detection level "very sparse growth" of microbes the sensitivity and specificity of MRS reached 68% and 75%, respectively. In an earlier study, following the same experimental protocol, but with calcium hydroxide as intracanal dressing, the corresponding values were 33% and 81%. In 25% of these cases bacteria persisted in the canals. As compared to calcium hydroxide, the use of IPI resulted in improved test accuracy, but loss of antibacterial capacity. Conclusively, intracanal dressings seem to vary in their influence on the microbiologic test performance as well as in their antibacterial efficacy. In a clinical situation the choice of interappointment dressing should include consideration of these potentially conflicting properties.     

Effect of endodontic procedures on enterococci, enteric bacteria and yeasts in primary endodontic infections

AIM: To detect enterococci, enteric bacteria and yeast species from the canals of teeth with primary endodontic infections before and after canal preparation and to test the antibiotic susceptibility of enterococcal strains isolated from infected root canals. METHODOLOGY: Twenty-five single-rooted teeth with pulp necrosis, intact pulp chambers and periradicular lesions were selected for study. Samples were collected from canals before and after instrumentation. Amongst isolated microorganisms from infected root canals only enterococci, enteric bacteria and yeasts were identified by biochemical tests. The in vitro antimicrobial susceptibility of isolated enterococci strains was evaluated by the Etest system. RESULTS: Microorganisms were isolated from 92% of the samples following intracoronal access, 22% were enterococci, enteric bacteria or yeast species. After biomechanical preparation, these species were no longer detected. After 7 days without intracanal dressing, 100% of the canals contained microorganisms, 52% of which were target species. However, after using paramonochlorophenol [PRP (2.0 g), Rinosoro and polyethylene glycol (400 equal parts up to 100 mL)] as an intracanal dressing for 7 days, enteric bacteria and yeasts were not detected; only enterococci were still present. All strains of enterococci were susceptible to ampicillin, but exhibited variable susceptibility to rifampin and ciprofloxacin. CONCLUSIONS: Enterococci, enteric bacteria and yeasts were present in primary endodontic infections. Enterococci, particularly Enterococcus faecalis and E. faecium were resistant to removal by root canal preparation followed by intracanal dressing.     

Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis

AIMS: The aim of this study was to determine the occurrence and role of yeasts, enteric gram-negative rods and Enterococcus species in root-filled teeth with chronic apical periodontitis, and the antimicrobial effect of iodine potassium iodide (IKI) irrigation. METHODOLOGY: Forty symptom-free root-filled teeth with chronic apical periodontitis were included in the study. The patients were divided into two groups. In group A the canals were filled with calcium hydroxide for 10-14 days after cleaning and shaping; in group B the canals were irrigated with IKI for 5 min after cleaning and shaping followed by a permanent root filling. Microbiological samples were taken from the canals before and after the chemomechanical preparation and after iodine irrigation (group B). RESULTS: Microbes were isolated from 33 of 40 teeth in the initial sampling. Yeasts were isolated from six teeth, three of them together with E. faecalis. Enteric rods (Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis) were present in three teeth and E. faecalis was isolated from 21 of the 33 culture positive teeth, 11 in pure culture. Growth was detected in 10 teeth of the second samples. Six of the 10 cases were E. faecalis, with five being a pure culture. All third samples (after IKI) except one were negative. The number of microbial cells per sample did not correlate with lesion size. Two flare-ups were recorded, both in teeth with a mixed infection. CONCLUSION: The high prevalence of enteric bacteria and yeasts in root-filled teeth with chronic apical periodontitis was established. IKI improved the antimicrobial effect of the treatment.     

Isolation of Enterococcus faecalis in previously root-filled canals in a Lithuanian population

The occurrence of Enterococcus faecalis in root canals of previously root filled teeth with apical periodontitis requiring retreatment was studied in Lithuanian patients. Twenty-five asymptomatic teeth were included in the study. Avoiding contamination microbiological samples were taken from the canals before and after preparation and irrigation with sodium hypochlorite and EDTA. Microbes were isolated from 20 of 25 teeth. E. faecalis was isolated from 14 of those 20 culture positive teeth, usually in pure culture or as a major component of the flora. Second samples taken after preparation revealed growth in 7 of the 20 teeth. Five of the seven cases were E. faecalis in pure culture. Isolation of E. faecalis was not related to the use of any particular root filling material in the original root filling. The results indicate that, rather than previous chemical treatment, it is the ecological conditions present in the incompletely filled root canal that are important for the presence of E. faecalis in these teeth.     

Microbial culture and checkerboard DNA-DNA hybridization assessment of bacteria in root canals of primary teeth pre- and post-endodontic therapy with a calcium hydroxide/chlorhexidine paste

International Journal of Paediatric Dentistry 2011 Aim. To investigate the root canal microbiota of primary teeth with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. Design. Baseline samples were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed for checkerboard DNA–DNA hybridization. Results. Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black‐pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically diminished compared to initial contamination (P <0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA–DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). Conclusion. Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing.     

Effects of extensive apical reaming and calcium hydroxide dressing on bacterial infection during treatment of apical periodontitis: a pilot study

An apical dentine sampling technique was applied in order to monitor the bacteriology of the pulp canal and radicular dentine before and during treatment of teeth with chronic apical periodontitis. Twenty-three teeth with a radiographic diagnosis of apical periodontitis were studied. They were subjected to a standardized two-appointment treatment regimen of extensive apical reaming in the absence of antimicrobial agents and 1-week dressing with calcium hydroxide. Bacteriological samples were taken from the root canal at the start, and apical dentine samples at the end, of each sitting. Provision was made to allow growth of anaerobic bacteria. All root canals but one showed growth at the start of treatment. Dentine samples were positive in 14 of the 23 teeth at the end of the first appointment. Eight of the 23 canals had detectable growth from the canal at the start of the second appointment, but in sufficient numbers for quantification in only one root canal. The subsequent dentine samples were otherwise negative at the second appointment. There was a tendency for teeth causing symptoms to harbour more bacteria than symptomless teeth.     

Circulating antibodies after experimental chronic infection in the root canal of teeth in monkeys

Abstract – Root canals of 49 teeth with necrotic pulp tissue in five monkeys were infected with Streptococcus faecalis ss liquefaciens, Actinornyces bovis, Fusobacteriam nucleatum, Peptostreptococcus anaerobiuS; and Bacteroidcs oralis in various combinations. After 6 months the root canals and the periapical tissues were subjected to radiographic., foacteriologic, and histologic examinations. Signs of periapical inflammation were radiographically registered in 41 teeth, most frequently in teeth inoculated with a mixed flora. In 11 out of 16 teeth infected separately with Strep, faecalis ss iiquefaciens, apical periodontitis was observed. Sera from the monkeys prior to and after the experimental inoculation were analyzed by means of gel diffusion, hemagghitination (HA), and complement binding test for antibodies against different antigen preparations of the homologous bacterial strains used for the inoculation. Detectable antibodies were seen with antigens of B. oralis in all five monkeys. Agglutinating antibodies were demonstrated with lipopolysaccharide-antigen of B. oralis in titers between 1:40 and 1:320. A marked reduction of the antibody level against lipopolysaccharide (LPS) antigen after mercaptoethanol treatment indicated that a main part of the antibodies registered was of IgM-class. The study shows that certain antigens of bacterial origin from infected root canals, while affecting the periapical tissues, also stimulate the production of circulating antibodies.     

Influence of infection at the time of root filling on the outcome of endodontic treatment of teeth with apical periodontitis

This study investigated the role of infection on the prognosis of endodontic therapy by following-up teeth that had had their canals cleaned and obturated during a single appointment. The root canals of 55 single-rooted teeth with apical periodontitis were thoroughly instrumented and irrigated with sodium hypochlorite solution. Using advanced anaerobic bacteriological techniques, post-instrumentation samples were taken and the teeth were then root-filled during the same appointment. All teeth were initially infected; after instrumentation low numbers of bacteria were detected in 22 of 55 root canals. Periapical healing was followed-up for 5 years. Complete periapical healing occurred in 94% of cases that yielded a negative culture. Where the samples were positive prior to root filling, the success rate of treatment was just 68%— a statistically significant difference. Further investigation of three failures revealed the presence of Actinomyces species in each case; no other specific bacteria were implicated in failure cases. These findings emphasize the importance of completely eliminating bacteria from the root canal system before obturation. This objective cannot be reliably achieved in a one-visit treatment because it is not possible to eradicate all infection from the root canal without the support of an inter-appointment antimicrobial dressing.     

Clinical investigation of the effect of calcium hydroxide intracanal dressing on bacterial lipopolysaccharide reduction from infected root canals

The purpose of this clinical study was to determine the effect of 7 day intracanal dressing with calcium hydroxide on the amount of bacterial lipopolysaccharide (LPS; endotoxin) in human teeth with necrotic and infected pulp and apical periodontitis. Twenty‐five single‐rooted teeth with necrotic pulps and apical periodontitis were selected. Samples were collected before (S1), after root canal preparation (S2) and after 7 day intracanal dressing with calcium hydroxide (S3). The limulus amoebocyte lysate assay was used to quantify LPS. LPS was present in 100% of the root canals before (S1), after preparation (S2) and after 7 day intracanal dressing (S3). A significant reduction, equal to 29.54%, was found after root canal preparation (P < 0.05). A significant difference (equal to 25.26% reduction) was also detected between S2 and S3 (P < 0.05). Total endotoxin reduction (S3 compared with S1) was found to be 47.34%. Endotoxin concentration of the infected root canals was reduced after root canal preparation and also after 7 days of dressing of canals with calcium hydroxide; however, relatively high values of endotoxin remained in the root canals.     

Isolation and identification of Enterococcus faecalis from necrotic root canals using multiplex PCR

This study was designed to survey the incidence of Enterococcus faecalis infection in symptomatic and asymptomatic root canals of necrotic teeth using PCR and to isolate the bacterium for further screening. Sixty patients categorized according to their clinical symptoms were used for sampling by insertion of paper points into the root canals and absorbing all the fluids present within them. The samples were incubated in 1.0 ml 2xYT (containing 16 g bacto tryptone, 10 g yeast extract and 5.0 g NaCl per liter) for 24 h at 37 degrees C without aeration prior to multiplex PCR analysis. To assist the isolation of E. faecalis, sub-samples were further grown in the same medium supplemented with 6.5% NaCl and back-inoculated into bile esculin. Using multiple cultivation-dependent and PCR analyses, 6 cases (10%) of E. faecalis were identified. Four isolates were obtained from asymptomatic cases of chronic apical periodontitis, and the other two were associated with phoenix abscess and acute apical abscess, respectively. No E. faecalis infection was found in 5 patients with acute apical periodontitis or in 9 with chronic suppurative periodontitis. Our results indicate that there is no significant difference in the incidence of E. faecalis between symptomatic and asymptomatic necrotic dental root canals (P > 0.05).     

血管内皮生长因子和骨形成蛋白2诱导犬恒牙原位牙髓再生

目的通过选择犬根尖孔发育完成的恒牙建立根尖周炎模型,探索血管内皮生长因子（VEGF）和骨形态生成蛋白2（BMP2）诱导原位牙髓再生的可能性。 方法2只10~ 12月龄的杂种犬,选择根尖孔发育完成的14颗恒前牙建立根尖周炎模型,分别将VEGF（VEGF组）、BMP2（BMP2组）单独和VEGF+BMP2联合（VEGF+BMP2组）与水凝胶复合植入感染控制后的根管腔内,对照组仅植入水凝胶。8周后组织学观察根管内组织再生情况。 结果植入8周后,VEGF组和VEGF+BMP2组根管腔内可见含有大量成纤维样细胞和血管的新生组织形成;而BMP2组和对照组根管腔内见均质状物质,未见细胞、血管形成。 结论VEGF或VEGF+BMP2复合水凝胶支架可以诱导犬根尖发育成熟的根尖周炎患牙在根管腔内生成含有血管的疏松结缔组织。     

Chemical and antimicrobial properties of calcium hydroxide mixed with irrigating solutions

AIM: Chemical and antimicrobial effects of paste-like suspensions of calcium hydroxide (Ca(OH)2) powder with chlorhexidine (CHx), sodium hypochlorite (NaOCl) or iodine potassium iodide (IPI) solutions were tested and compared to conventional Ca(OH)2/saline paste. METHODOLOGY: Equivalent mixtures of Ca(OH)2 with irrigating solutions were compared to conventional Ca(OH)2/saline paste for their ability to raise the pH in root dentine. Ca(OH)2 pastes were compacted into prepared root canals of single-rooted extracted human teeth. Changes in root pH over time were recorded with a microelectrode placed in standardized wells within root surfaces. Antimicrobial efficacy of test solutions and pastes against Enterococcus faecalis and Candida albicans was assayed using an agar diffusion test. RESULTS: The ability of Ca(OH)2 to raise the pH in the root dentine was maintained when using mixtures of Ca(OH)2 with CHx, NaOCl or IPI (Kruskal-Wallis, P > 0.05). Mixtures of Ca(OH)2 with the test solutions had similar antimicrobial properties as equivalent mixtures with saline (anova, P > 0.05). The efficacy of CHx was reduced when mixed with Ca(OH)2. No additive antiseptic effects were recorded between Ca(OH)2 and IPI or Ca(OH)2 and NaOCl. CONCLUSIONS: Ca(OH)2 is a strong alkali, whose pH is not changed when adding weak acids or alkalis in aqueous suspensions. Under the conditions of this study, mixing Ca(OH)2 powder with the test irrigating solutions did not provide an increased antimicrobial effect compared to a conventional Ca(OH)2/saline medication.     

Antimicrobial substantivity of chlorhexidine-treated bovine root dentin

Previous studies have demonstrated antimicrobial substantivity in root canal dentin up to 7 days after treatment with chlorhexidine. This in vitro study assessed the antimicrobial substantivity of chlorhexidine-treated bovine root dentin over a period of 21 days. Sixty standardized bovine root sections were randomly divided into three equal groups, and their canals immersed in one of the following solutions: (i) sterile saline; (ii) 2.5% NaOCl; or (iii) 0.2% chlorhexidine (CHX). Half the specimens in each group were treated with the solution for 5 min and the other half for 7 days. After solutions were removed, the specimens were incubated at 37 degrees C in Brain Heart Infusion broth containing Enterococcus faecalis (ATCC 29212). A fresh inoculum was added to the broth every other day over a 21-day period. The canals were then enlarged with sterile burs, and the dentin shavings collected and cultured for the presence of cultivable bacteria in the dentinal tubules. Specimens treated with CHX for 7 days demonstrated significantly less dentin colonization by E. faecalis than the other specimens. CHX has potential as an intracanal medicament, if it can be applied for a period of at least 7 days.     

Bacteria recovered from teeth with apical periodontitis after antimicrobial endodontic treatment

Chávez de Paz LE, Dahlén G, Molander A, Möller Å, Bergenholtz G . Bacteria recovered from teeth with apical periodontitis after antimicrobial endodontic treatment. International Endodontic Journal, 36 , 500–508, 2003. Aim To determine whether there is a pattern for certain bacteria to remain after chemo‐mechanical treatment of root canals in teeth with apical periodontitis. Methodology Consecutive root‐canal samples of 200 teeth receiving root‐canal treatment, referred from general practitioners and endodontic specialists for analyses of cultivable microbes, were studied prospectively. To be included, samples had to be taken at a treatment session subsequent to the one at which endodontic therapy was initiated. All samples were from teeth that either presented with clinical or radiographic evidence of apical periodontitis or both. Bacteriological findings were linked to clinical and radiographic parameters including status of the root canal prior to treatment, namely, vital pulp, necrotic pulp or root filled. Results A total of 248 strains were isolated from 107 teeth giving bacterial growth. Gram‐positives predominated (85%). Lactobacillus spp. (22%), nonmutans streptococci (18%), and Enterococcus spp. (12%) were the most common isolates. Gram‐negative anaerobes were relatively sporadic. Large radiographic bone lesions, persistent pain and use of intracanal calcium hydroxide dressing correlated with bacterial presence (P < 0.05). Conclusions Once established, nonmutans streptococci, enterococci and lactobacilli appear to survive commonly following root‐canal treatment of teeth with clinical and radiographical signs of apical periodontitis.     

Radiographic evaluation of periapical healing of permanent teeth with periapical lesions after extrusion of AH Plus sealer

The aim of this study was to clinically and radiographically examine the effects of extrusion of AH Plus sealer on the healing of permanent teeth with apical periodontitis. A total of 87 root canals radiographically detected with apical periodontitis were included in the study. Posttreatment radiographs indicated sealer extrusion into 49 canals (Group 1) and no sealer extrusion into 38 canals (Group 2). Periapical treatment was judged as complete healing (CH), incomplete healing (IH) and no healing (NH) at the end of a 4-year follow-up period. Amounts of extraradicular sealer were recorded as "unchanged," "reduced," "almost absent," or "absent." The t test was used for the statistical analyses. In Group 1, CH was detected in 41 canals, IH in 4 canals, and NH in 4 canals. Differences between CH and both IH and NH were statistically significant (P < .001). In Group 2, CH was detected in 34 canals and NH in 4 canals. The difference between CH and NH was statistically significant (P < .001). A statistically significant difference (P < .05) between treatment groups was observed for CH at the 6-month follow-up appointment only; other than that instance, there were no statistical differences for CH or NH between the groups. In conclusion, extruded AH Plus does not prevent periapical healing, but can be a delaying factor for healing in children.     

Antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in infected human root canals

AIM: To determine the antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in human root canals infected by Enterococcus faecalis. METHODOLOGY: Thirty human maxillary anterior teeth were prepared and inoculated with E. faecalis for 60 days. Eppendorf tubes were connected to the coronal portion of the teeth. Urethane hoses were attached to the tubes and to the entrance of a peristaltic pump. The exit of the apparatus corresponded to the apical portion of the root canals. The test irrigating solutions were ozonated water, gaseous ozone, 2.5% sodium hypochlorite (NaOCl), 2% chlorhexidine that circulated at a constant flow of 50 mL min(-1) for 20 min. Samples from the root canals were collected and immersed in 7 mL Letheen Broth (LB), followed by incubation at 37 degrees C for 48 h. Bacterial growth was analysed by turbidity of the culture medium and subculture on a specific nutrient broth. A 0.1 mL inoculum obtained from LB was transferred to 7 mL of brain heart infusion and incubated at 37 degrees C for 48 h. Bacterial growth was checked by turbidity of the culture medium carried out in triplicate. RESULTS: No solution used as an irrigant over a 20-min contact time demonstrated an antimicrobial effect against E. faecalis. CONCLUSION: The irrigation of infected human root canals with ozonated water, 2.5% NaOCl, 2% chlorhexidine and the application of gaseous ozone for 20 min was not sufficient to inactivate E. faecalis.     

In vitro evaluation of the effectiveness of the chemomechanical preparation against Enterococcus faecalis after single- or multiple-visit root canal treatment

The purpose was to assess the elimination of Enterococcus faecalis in vitro in human mandibular premolars after chemomechanical preparation with or without the use of a calcium hydroxide dressing. After 60 days of contamination with E. faecalis, the root canals were prepared using the Crown-Down technique combined with 2% chlorhexidine gel irrigation. Then, the specimens were divided into two experimental groups, treated in a single visit or in multiple visits, and two control groups. The multiple-visit group received a dressing with calcium hydroxide for 14 days (Calen) and the single-visit group did not receive any medication. In the two control groups, the canals were filled with BHI after chemomechanical preparation with 2% chlorhexidine gel or distilled water. Microbial samples were taken from the root canals for colony forming unit count for each phase of the treatment using sterile paper points inside the root canal lumen. Data were ranked and analyzed by the Kruskal-Wallis statistical test. The residual microbial colonies were then assessed. The results showed that chemomechanical preparation using 2% chlorhexidine gel with no intra-canal dressing reduced by 100% the E. faecalis contamination of the root canal lumen. The calcium-hydroxide group that received the 14-day intra-canal dressing allowed a small number of bacteria to grow between visits, but without statistical differences between groups.     

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.