Effect of root canal procedures on endotoxins and endodontic pathogens

BACKGROUND/AIMS: The purpose of this study was to determine the amount of endotoxin (lipopolysaccharide) and cultivable bacteria in human necrotic root canals before (S1) and after chemo-mechanical preparation using chlorhexidine (CHX) gel as auxiliary chemical substance (S2), and after 7 days of intracanal dressing (S3) in order to evaluate the anti-endotoxin and antimicrobial effects of endodontic procedures. METHOD: Twenty-four teeth were selected for the present study. Chemo-mechanical preparation was performed using 2% CHX gel and three different intracanal medicaments [CaOH2 paste; 2% CHX gel; and CaOH2 + 2% CHX gel]. A quantitative chromogenic Limulus amoebocyte lysate assay was used to measure the amount of endotoxin. Aerobic and anaerobic techniques were used to isolate and identify bacteria, and to determine the bacterial reduction by counting colony-forming units (CFU). RESULTS: Endotoxins and bacteria were present in 100% of the initial samples, with endotoxin concentration ranging from 62.93 to 214.56 UE/ml and CFU ranging from 4 x 10(5) to 2.6 x 10(6). After chemo-mechanical preparation a mean endotoxin reduction of 44.4% was found. Eight (33.3%) root canals were still positive by culture analysis with a mean reduction of bacteria (CFU) of 99.96%. After 7 days of intracanal dressing, endotoxin concentration decreased by only 1.4% compared with S2, and residual bacteria were recovered by culture analysis in 13 cases (54.1%). No significant difference was found among different intracanal medicaments. CONCLUSION: Relatively high values of endotoxin were still present in the root canal after chemo-mechanical preparation although the majority of bacteria were eliminated. No improvement was achieved by 7 days of intracanal dressing.     

Reduction in the cultivable bacterial populations in infected root canals by a chlorhexidine-based antimicrobial protocol

The present clinical study was conducted to assess the bacterial reduction after chemomechanical preparation using 0.12% chlorhexidine digluconate solution as an irrigant and the additive antibacterial effect of intracanal dressing with calcium hydroxide (Ca(OH)(2)) associated with 0.12% chlorhexidine digluconate gel. According to stringent inclusion/exclusion criteria, 13 teeth with primary intraradicular infections and chronic apical periodontitis were selected and followed in the study. Bacterial samples were taken at the baseline (before treatment) (S1), after chemomechanical preparation using chlorhexidine (CHX) as an irrigant (S2), and after a 7-day dressing with Ca(OH)(2)/CHX paste (S3). Cultivable bacteria recovered from infected root canals at the three stages were counted and identified by means of 16S ribosomal RNA gene sequencing analysis. At S1, all canals were positive for bacteria, with the mean number of 3.5 taxa per canal (range, 2-9). At S2, 7 cases (53.8%) still harbored cultivable bacteria, with a mean number of 1.7 taxon per canal (range, 1-4). At S3, only one case (7.7%) was positive for the presence of bacteria. The great majority of taxa found in posttreatment samples were gram-positive bacteria. A significantly high reduction in bacterial counts was observed between S1 and S2 and S1 and S3 (p<0.001). Also, significant differences were observed for comparisons involving S2 and S3 samples with regard to both quantitative bacterial reduction (p=0.014) and number of cases yielding negative cultures (p=0.01). It was concluded that chemomechanical preparation with 0.12% CHX solution as an irrigant significantly reduced the number of intracanal bacteria but failed to render the canal free of cultivable bacteria in about one half of the cases. Application of a 7-day intracanal dressing with Ca(OH)(2)/CHX paste further increased significantly the number of cases yielding negative cultures.     

Comparative analysis of endodontic pathogens using checkerboard hybridization in relation to culture

Background/Aim:  The purpose of this study was to detect bacterial species and to quantify the total number of bacteria from samples of infected root canals before (S1) and after chemo-mechanical preparation using 2% chlorhexidine (CHX) gel as auxiliary chemical substance (S2) and after 7 days of intracanal dressing (S3) to compare microbial changes.
Method:  Twenty-four teeth were selected for this study. Chemo-mechanical preparation was performed using 2% CHX gel, then three different intracanal medicaments [M1: Ca(OH)₂ paste; M2: 2% CHX gel; and M3: Ca(OH)₂ paste plus 2% CHX gel] were used for 7 days. Checkerboard DNA–DNA hybridization was performed to detect 40 bacterial species. Aerobic and anaerobic culture techniques were used to determine the bacterial community by counting the colony-forming units (CFU).
Results:  The species most frequently identified by checkerboard in S1 were: Fusobacterium nucleatum ssp . polymorphum , Treponema socranskii ssp. socranskii , Parvimonas micra and Enterococcus faecalis. In S2 and S3 a total of eight different species were identified; and only one of them was gram-positive ( E. faecalis ). Microorganisms were not identified after use of M2 for 7 days. The quantification obtained on agar plates ranged from 4 × 10⁵ to 2.6 × 10⁶ CFU/ml in S1, mean CFU was reduced by 99.96% in S2, and there was no statistical difference between the CFU in S2 and S3.
Conclusion:  The antibacterial effect of the mechanical preparation supplemented by the use of an antibacterial auxiliary substance greatly reduced the microorganisms in the main root canal.     

Bacterial reduction in infected root canals treated with 2.5% NaOCl as an irrigant and calcium hydroxide/camphorated paramonochlorophenol paste as an intracanal dressing

This clinical study investigated the bacterial reduction after instrumentation using 2.5% sodium hypochlorite (NaOCl) as an irrigant and further interappointment dressing with a calcium hydroxide (Ca(OH)(2))/camphorated paramonochlorophenol (CPMC) paste. Eleven teeth with primary intraradicular infections and chronic apical periodontitis selected according to stringent inclusion/exclusion criteria followed in the study. Bacterial samples were taken before treatment (S1), after chemomechanical preparation using hand NiTi files and 2.5% NaOCl (S2), and following a 7-day medication with a Ca(OH)(2) paste in CPMC (S3). Cultivable bacteria recovered from infected root canals at the three stages were counted and identified by means of 16S rRNA gene sequencing analysis. At S1, all cases harbored bacteria, with a mean number of 2.8 taxa per canal (range, 1-6). At S2, 6 of 11 (54.5%) of the cases yielded positive cultures, with one to three species per canal. At S3, only one case (9.1%) was positive for the presence of bacteria, with Propionibacterium acnes as the only taxon isolated. A significantly high reduction in bacterial counts was observed between S1 and S2, and S1 and S3. Significant differences were also observed for comparisons involving S2 and S3 samples with regard to both quantitative bacterial reduction (p = 0.029) and number of culture-negative cases (p = 0.03). It was concluded that chemomechanical preparation with 2.5% NaOCl as an irrigant significantly reduced the number of bacteria in the canal but failed to render the canal free of cultivable bacteria in more than one-half of the cases. A 7-day intracanal dressing with Ca(OH)(2)/CPMC paste further significantly increased the number of culture-negative cases.     

Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis

Objective:

The aim of this study was to monitor the effectiveness of root canal procedures by using different irrigants and intracanal medication on endotoxin levels found in root canals of teeth with chronic apical periodontitis.

Material and Methods:

Thirty root canals of teeth with pulpal necrosis associated with periapical lesions were selected and randomly divided into groups according to the irrigants used: GI - 2.5% NaOCl, GII - 2% chlorhexidine (CHX) gel, and GIII - saline solution (SS) (all, n=10). Samples were collected with sterile/apyrogenic paper points before (S1) and after root canal instrumentation (S2), after use of 17% ethylenediaminetetraacetic acid (EDTA) (S3), and after 30 days of intracanal medication (Ca(OH)₂+SS) (S4). A turbidimetric kinetic Limulus Amebocyte Lysate assay was used for endotoxin measurement.

Results:

Endotoxins were detected in 100% of the root canals investigated (30/30), with a median value of 18.70 EU/mL. After S2, significant median percentage reduction was observed in all groups, irrespective of the irrigant tested: 2.5% NaOCl (99.65%) (GI), 2% CHX (94.27%) (GII), and SS (96.79%) (GIII) (all p<0.05). Root canal rinse with 17% EDTA (S3) for a 3-minute period failed to decrease endotoxin levels in GI and a slight decrease was observed in GII (59%) and GIII (61.1%) (all p>0.05). Intracanal medication for 30 days was able to significantly reduce residual endotoxins: 2.5% NaOCl (90%) (GI), 2% CHX (88.8%) (GII), and SS (85.7%) (GIII, p<0.05). No differences were found in the endotoxin reduction when comparing s2 and s4 treatment groups.

Conclusion:

Our findings demonstrated the effectiveness of the mechanical action of the instruments along with the flow and backflow of irrigant enduring root canal instrumentation for the endotoxin removal from root canals of teeth with chronic apical periodontitis. Moreover, the use of intracanal medication for 30 days contributed for an improvement of endotoxin reduction.     

Bacterial quantification in teeth with apical periodontitis related to instrumentation and different intracanal medications: a randomized clinical trial

The antibacterial efficacy of intracanal medication with calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX), and a combination of both [Ca(OH)2/CHX] was assessed in teeth with chronic apical periodontitis. Thirty-three canals were instrumented, randomly divided into three groups, and medicated with either Ca(OH)2, CHX, or Ca(OH)2/CHX. Bacteriological samples obtained from the operative field and the root canals before (S1) and after instrumentation (S2) in the first treatment session, and after medication (S3) in the second session 1 week later, were assessed for bacterial growth, observed by turbidity and in agar plates, and viable colony-forming unit (CFU) counts. Bacterial growth and CFU counts decreased significantly from S1 to S2 (Mann-Whitney, p<0.05). Differences in growth and counts between S2 to S3 were not statistically significant for all three intracanal medication groups. It was concluded that the antibacterial efficacy of Ca(OH)2, CHX, and Ca(OH)2/CHX was comparable.     

Effectiveness of castor oil extract on Escherichia coli and its endotoxins in root canals

This in vitro study sought to evaluate the effectiveness of castor oil extract used as an irrigating solution on Escherichia coli and its endotoxins in root canals. Sixty single-rooted teeth were prepared (using castor oil extract as irrigating solution) and divided into five groups (n = 12): Group 1 samples were treated with calcium hydroxide (Ca(OH)2), Group 2 samples were treated with polymyxin B, Group 3 samples were treated with Ca(OH)2 and 2% chlorhexidine gel (CHX), and Group 4 samples were treated with castor oil extract. A control group used physiological saline solution as an irrigant. Canal content samples were collected at four different times: immediately after instrumentation, seven days after instrumentation, after 14 days of intracanal medication, and seven days after removal of intracanal medication. A plating method was used to assess antimicrobial activity and the quantification of endotoxins was evaluated by the chromogenic Limulus lysate assay. Data were submitted to ANOVA and a Dunn test (a = 5%). Irrigation with castor oil extract decreased E. coli counts but had no effect on the level of endotoxins. Samples taken seven days after removal of medication revealed a significant reduction in endotoxin levels in Groups 3 and 4. Compared to the saline solution irrigation, castor oil extract decreased microorganism counts in root canals immediately after canal preparation. None of the medications used completely eliminated endotoxins in the root canal.     

Bacteriologic evaluation of ultrasonic root canal instrumentation

The antibacterial effect of ultrasonic instrumentation in the treatment of infected root canals was clinically evaluated. Sodium hypochlorite solution (0.5%) was used as an irrigant, but no antibacterial intracanal dressing was used between the appointments. The ultrasonic technique eliminated the bacteria from the canals more efficiently than hand instrumentation alone. Even though ultrasonication definitely improves the procedure of root canal disinfection, the use of an antibacterial dressing between appointments is necessary to achieve as complete a reduction in bacterial levels as possible.     

Macrophage Cell Activation with Acute Apical Abscess Contents Determined by Interleukin-1 Beta and Tumor Necrosis Factor Alpha Production

Introduction

This clinical study has investigated the antigenic activity of bacterial contents from exudates of acute apical abscesses (AAAs) and their paired root canal contents regarding the stimulation capacity by levels of interleukin (IL)-1 beta and tumor necrosis factor alpha (TNF-α) throughout the root canal treatment against macrophage cells.

Methods

Paired samples of infected root canals and exudates of AAAs were collected from 10 subjects. Endodontic contents were sampled before (root canal sample [RCS] 1) and after chemomechanical preparation (RCS2) and after 30 days of intracanal medication with calcium hydroxide + chlorhexidine gel (Ca[OH]₂ + CHX gel) (RCS3). Polymerase chain reaction (16S rDNA) was used for detection of the target bacteria, whereas limulus amebocyte lysate was used to measure endotoxin levels. Raw 264.7 macrophages were stimulated with AAA exudates from endodontic contents sampled in different moments of root canal treatment. Enzyme-linked immunosorbent assays were used to measure the levels of TNF-α and IL-1 beta.

Results

Parvimonas micra, Porphyromonas endodontalis, Dialister pneumosintes, and Prevotella nigrescens were the most frequently detected species. Higher levels of endotoxins were found in samples from periapical exudates at RCS1 (P < .005). In fact, samples collected from periapical exudates showed a higher stimulation capacity at RCS1 (P < .05). A positive correlation was found between endotoxins from exudates with IL-1 beta (r = 0.97) and TNF-α (r = 0.88) production (P < .01). The significant reduction of endotoxins and bacterial species achieved by chemomechanical procedures (RCS2) resulted in a lower capacity of root canal contents to stimulate the cells compared with that at RCS1 (P < .05). The use of Ca(OH)₂ + CHX gel as an intracanal medication (RCS3) improved the removal of endotoxins and bacteria from infected root canals (P < .05) whose contents induced a lower stimulation capacity against macrophages cells at RCS1, RCS2, and RCS3 (P < .05).

Conclusions

AAA exudates showed higher levels of endotoxins and showed a greater capacity of macrophage stimulation than the paired root canal samples. Moreover, the use of intracanal medication improved the removal of bacteria and endotoxins from infected root canals, which may have resulted in the reduction of the inflammatory potential of the root canal content.     

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.     

Effects of chemomechanical preparation with 2.5% sodium hypochlorite and intracanal medication with calcium hydroxide on cultivable bacteria in infected root canals

This clinical study was conducted to assess the bacterial reduction after chemomechanical preparation with 2.5% NaOCl as an irrigant and the additive antibacterial effect of intracanal dressing with calcium hydroxide. According to stringent inclusion criteria, 11 teeth with primary intraradicular infections and chronic apical periodontitis were selected and monitored in the study. Bacterial samples were taken at the baseline (before treatment) (S1), after chemomechanical preparation with 2.5% NaOCl as an irrigant (S2), and after a 7-day dressing with a calcium hydroxide paste in glycerin (S3). Cultivable bacteria recovered from infected root canals at the 3 stages were counted and identified by means of 16S rRNA gene sequencing analysis. At S1, all canals were positive for bacteria, with the mean number of 2.8 taxa per canal (range, 1-6). At S2, 5 cases (45.5%) still harbored cultivable bacteria, with 1 or 2 species per canal. At S3, bacteria were cultured from 2 cases (18.2%), with 1 species per positive case. There was no indication that any specific bacterial taxon was more resistant to treatment. A significant reduction in bacterial counts was observed between S1 and S2, and S1 and S3. However, no statistically significant difference was observed for comparisons involving S2 and S3 samples with regard to the number of cases yielding negative cultures (P = .18) or quantitative bacterial reduction (P = .19). It was concluded that the whole antibacterial protocol used in this study significantly reduced the number of bacteria in the canal and rendered most canals free of cultivable bacteria.     

Effect of calcium hydroxide intracanal dressing on the bond strength of a resin-based endodontic sealer

The aim of this in vitro study was to evaluate the bond strength of Epiphany resin-based sealer to dentin walls after placement of calcium hydroxide [Ca(OH)2] dressings. Fifteen extracted single-rooted human teeth were instrumented using 2.5% NaOCl + EDTA as irrigants. The teeth were randomly assigned to 3 groups (n=5), according to the intracanal dressing: G1= Ca(OH)2 + saline; G2= Ca(OH)2 + 2% chlorhexidine gluconate (CHX) gel; and G3= saline (control). After 10 days of storage in 100% humidity at 37 degrees C, the dressings were removed and the root canals were filled with Epiphany sealer. After additional 48 h of storage, the specimens were sectioned transversally into 2-mm-thick discs. Push-out tests were performed (1 mm/min, Instron 4411) and the maximum loads at failure were recorded in MPa. One-way ANOVA and Newman-Keuls tests showed a statistically significant decrease in bond strength when a Ca(OH)2 dressing was used before root canal filling with Epiphany (G1= 10.18 +/- 1.99 and G2= 9.98 +/- 2.97) compared to the control group (13.82 +/- 3.9) (p< 0.05). It may be concluded that the use of Ca(OH)2 as an intracanal dressing material affected the adhesion of Epiphany to the root canal walls, but even though the values were within the acceptable range found in the literature.     

Detection and eradication of microorganisms in root-filled teeth associated with periradicular lesions: an in vivo study

This study determined the presence of microorganisms by culture and polymerase chain reaction in asymptomatic root-filled teeth with periradicular lesions. Furthermore, a disinfecting regimen using sodium hypochlorite (NaOCl), ethylenediaminetetraacetic acid (EDTA), chlorhexidine digluconate (CHX) irrigation, and calcium hydroxide (Ca(OH)(2)) dressing was assessed. After removal of the root-filling material, specimens of 20 cases undergoing retreatment were sampled. Moreover, the canals were sampled after each step of the disinfecting regimen. Prevalence of microorganisms was 60% by culture and 65% by polymerase chain reaction. In four of those samples (31%), DNA of Enterococcus faecalis was found. After further root canal preparation and irrigation using NaOCl and EDTA, microorganisms could be detected in none of the teeth. Thus, CHX and Ca(OH)(2) could not show further disinfection. In contrast, microorganisms were found in two teeth after the interappointment dressing. It may be concluded that proper root canal preparation and irrigation using NaOCl and EDTA are sufficient for decontamination of the root canal system during endodontic retreatment.     

ANTIMICROBIAL ACTIVITY OF SODIUM HYPOCHLORITE ASSOCIATED WITH INTRACANAL MEDICATION FOR Candida albicans AND Enterococcus faecalis INOCULATED IN ROOT CANALS

Objective:

The purpose of this study was to evaluate the action of sodium hypochlorite (NaOCl) associated with an intracanal medication against Candida albicans and Enterococcus faecalis inoculated in root canals.

Material and Methods:

Thirty-six human single-rooted teeth with single root canals were used. The canals were contaminated with C. albicans and E. faecalis for 21 days and were then instrumented with 1% NaOCl. The roots were divided into 3 groups (n=12) according to the intracanal medication applied: calcium hydroxide paste, 2% chlorhexidine (CHX) gel, and 2% CHX gel associated with calcium hydroxide. The following collections were made from the root canals: a) initial sample (IS): 21 days after contamination (control), b) S1: after instrumentation, c) S2: 14 days after intracanal medication placement; S3: 7 days after intracanal medication removal. The results were analyzed statistically by the Kruskal-Wallis test at 5% significance level.

Results and Conclusions:

Both 1% NaOCl irrigation and the intracanal medications were effective in eliminating E. faecalis and C. albicans inoculated in root canals.     

Effectiveness of calcium hydroxide-based intracanal medication on infectious/inflammatory contents in teeth with post-treatment apical periodontitis

Objective

The objective of this work was to investigate in vivo the effects of calcium hydroxide-based intracanal medication (ICM) on the levels of bacteria, pro-inflammatory cytokines (PICs), and matrix metalloproteinases (MMPs) in root canals and periradicular tissues of teeth with failure of the root canal treatment and apical periodontitis.

Materials and methods

Twenty infected root canals of single-rooted teeth were randomly assigned into two groups according to the irrigant used for chemomechanical preparation (CMP) (n = 10 per group): G1 – 2% chlorhexidine (CHX) gel and G2 – 6% sodium hypochlorite (NaOCl). Root canal contents were taken by using paper points before CMP (S1) and after 30 days of calcium hydroxide-based ICM (S2). Microbial reduction was calculated by means of colony-forming unit count (CFU/mL), with PICs and MMPs (pg/mL) being measured by using enzyme-linked immunosorbent assay (ELISA).

Results

Culturable bacteria (101.2 ± 79.2), PICs (IL-1β 1.2 ± 0.4 and TNF-α 8.8 ± 4.7), MMP-2 (803.7 ± 96.4), MMP-3 (453.9 ± 229.3), MMP-8 (245.9 ± 122.4), MMP-9 (129.4 ± 29.6), and MMP-13 (70.8 ± 12.8) were present in all S1 samples. After 30 days of ICM (S2), a 99.5% microbial reduction was observed, together with a significant reduction of PICs in all groups. Overall, it was observed a decrease in the levels of MMPs (S2), except MMP-13, which was found in increased levels after ICM (P < .05), independently of the groups.

Conclusions

Calcium hydroxide-based intracanal medications have had a positive effect on the microbial reduction by decreasing the levels of PICs and MMPs. Both auxiliary chemical substances (i.e., 2% CHX and 6% NaOCl) presented similar effects when calcium hydroxide was used as intracanal medication.

Clinical relevance

Teeth with failure of the root canal treatment and apical periodontitis, and consequently with high levels of bacteria, PIC, and MMP, may present a better prognosis after a 30 days of a calcium hydroxide-based ICM.

     

Antibacterial efficacy of chlorhexidine gluconate intracanal medication in vivo

The antibacterial efficacy of intracanal medication with 2% chlorhexidine liquid (CHX) was assessed in teeth with apical periodontitis. Canals in 22 teeth were instrumented at the first session, medicated with CHX, and reaccessed after 7 to 15 days. Bacteriological samples were aspirated at the first and second sessions, before (1A, 2A) and after (1B, 2B) canal instrumentation. Viable bacterial counts were obtained by culture (CFU) and microscopy using vital dyes. Microscopic counts were higher than CFU counts. Consistently high CFU counts in 1A samples (mean, 2 x 10(5) microL(-1) canal volume) decreased significantly (p < 0.0001) in 1B samples, increased significantly (p < 0.04) in 2A samples, and decreased in 2B samples to the level of 1B samples. Proportions of negative cultures followed the pattern of CFU counts. Intracanal medication with CHX did not reduce the bacterial concentration. Bacterial counts expressed per microliter canal volume added information beyond the counts per tooth as expressed in previous studies.     

The effectiveness of three instrumentation techniques on the elimination of Enterococcus faecalis from a root canal: an in vitro study

The in vitro reduction of a bacterial population in a root canal by mechanical instrumentation using three techniques was evaluated. Root canals inoculated with a Enterococcus faecalis (E. faecalis) suspension were instrumented using hand Hedstroem files, Giromatic files, and Hero 642 rotary instruments. Irrigation was performed using sterile saline solution. Root canals were sampled before and after instrumentation. After serial dilutions, samples were plated onto Mitis-Salivarius agar and the colony forming units grown were counted. All instruments tested were able to significantly reduce the number of bacterial cells in the root canal, however, the results of this study indicated that Hedstroem files, Giromatic, and Hero 642 techniques were not significantly different in their ability to reduce intracanal bacteria.     

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.     

Reduction of intracanal bacteria using GT rotary instrumentation, 5.25% NaOCl, EDTA, and Ca(OH)2

This study was conducted to determine the bacterial reduction using Profile GT files and a strict irrigation protocol utilizing 5.25% NaOCl and EDTA. The additive antibacterial effect of Ca(OH)2 was also evaluated. In addition, the study compared the bacterial reduction with the GT protocol versus larger instrumentation. Thirty-one subjects with apical periodontitis were recruited. Bacterial samples were taken upon access (S1), after instrumentation and a strict irrigation protocol (S2), and following >1 wk of Ca(OH)2 (SC). A log10 transformation of colony forming units was done since sample bacterial counts are not normally distributed. At S1, 93.55% of canals sampled bacteria. At S2, 52.72% of the cases sampled bacteria. At SC, 14% of the cases cultured bacteria. The McNemar test showed a significant reduction (p<0.0009) in bacteria between S1 and S2. This was also true between S2 and SC (p<0.0019). It was concluded the GT protocol significantly reduced the number of bacteria in the canal but failed to render the canal bacteria free in more than half of the cases Ca(OH)2 application significantly further reduced bacteria. Lastly, large apical instrumentation removed more bacteria than small apical instrumentation.