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.     

Clinical efficiency of 2% chlorhexidine gel in reducing intracanal bacteria

This study evaluated the clinical efficacy of 2% chlorhexidine (CHX) gel on intracanal bacteria reduction during root canal instrumentation. The additional antibacterial effect of an intracanal dressing (Ca[OH](2) mixed with 2% CHX gel) was also assessed. Forty-three patients with apical periodontitis were recruited. Four patients with irreversible pulpitis were included as negative controls. Teeth were instrumented using rotary instruments and 2% CHX gel as the disinfectant. Bacterial samples were taken upon access (S1), after instrumentation (S2), and after 2 weeks of intracanal dressing (S3). Anaerobic culture was performed. Four samples showed no bacteria growth at S1, which were excluded from further analysis. Of the samples cultured positively at S1, 10.3% (4/39) and 8.3% (4/36) sampled bacteria at S2 and S3, respectively. A significant difference in the percentage of positive culture between S1 and S2 (p < 0.001) but not between S2 and S3 (p = 0.692) was found. These results suggest that 2% CHX gel is an effective root canal disinfectant and additional intracanal dressing did not significantly improve the bacteria reduction on the sampled root canals.     

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.     

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.     

In vivo evaluation of microbial reduction after chemo-mechanical preparation of human root canals containing necrotic pulp tissue

AIM: To determine in vivo, the degree of microbial reduction after chemo-mechanical preparation of human root canals containing necrotic pulp tissue when using two endodontic irrigating reagents, sodium hypochlorite (NaOCl) or chlorhexidine gel (CHX). METHODOLOGY: Thirty-two single rooted teeth with necrotic pulp were divided into two groups. One group (n=16) was irrigated with 2.5% NaOCl, whilst the other group (n=16) was irrigated with 2% CHX gel. Assessment of the bacterial load was accomplished by use of real-time quantitative-polymerase chain reaction (RTQ-PCR) directed against the small subunit ribosomal DNA using the SYBRGreen and TaqMan formats. Statistical analysis was performed using the Mann-Whitney test. For contrast, bacterial load was also determined by traditional culture techniques. RESULTS: The bacterial load was reduced substantially in both groups (over 96%). However, using RTQ-PCR the bacterial load before and after chemo-mechanical preparation was greater when compared with evaluation using colony forming units (CFU). Furthermore, as measured by RTQ-PCR, the bacterial reduction in the NaOCl-group (SYBRGreen 99.99%; TaqMan: 99.63%) was significantly greater (P<0.01) than in the CHX-group (SYBRGreen 96.62%; TaqMan: 96.60%). According to culture technique 75% of cases were free of bacteria after chemo-mechanical preparation in the NaOCl-group, whilst 50% of cases were bacteria free in the CHX-group. CONCLUSION: NaOCl has not only a higher capacity to kill microorganisms but is also more able to remove cells from the root canal.     

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.     

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.     

In vitro evaluation of the effectiveness of irrigants and intracanal medicaments on microorganisms within root canals

AIM: To evaluate in vitro the effectiveness of sodium hypochlorite (NaOCl), chlorhexidine (CHX) and five intracanal medicaments on microorganisms within root canals. METHODOLOGY: Ninety-six human single-rooted extracted teeth were used. After removing the crowns, canal preparation was completed and the external root surfaces were coated with epoxy resin. Following sterilization, the teeth were contaminated with Candida albicans and Enterococcus faecalis, and were incubated at 37 +/- 1 degrees C for 7 days. The teeth were divided according to the irrigant solution or intracanal medicament: group 1, sterile physiologic solution (SPS) and calcium hydroxide (Ca(OH)2) paste; group 2, SPS and camphorated paramonochlorophenol (CPMC); group 3, SPS and tricresol formalin; group 4, SPS and CaOH2 + CPMC paste; group 5, SPS and PMC furacin; group 6, 2.5% NaOCl without intracanal medication; group 7, 2.0% CHX without intracanal medication and group 8, SPS without intracanal medication (control group). Microbiological samples were collected with sterile paper points, and bacterial growth was determined. The data were submitted to the analysis of variance (anova, P = 0.05). RESULTS: For C. albicans, groups 3 and 8 were statistically less effective than groups 1, 2, 4 and 5 (Kruskal-Wallis (K-W) = 65.241; gl = 7; P = 0.001). For E. faecalis, groups 6 and 8 were statistically less effective than groups 1-4 and 7 (K-W = 61.048; gl = 7; P = 0.001). CONCLUSIONS: Ca(OH)2 + CPMC paste was the most effective intracanal medicament for the elimination of the two microorganisms; 2.0% CHX solution was more effective than 2.5% NaOCl against E. faecalis.     

Microbial analysis of endodontic infections in teeth with post-treatment apical periodontitis before and after medication

This study aimed to determine the intraradicular microbiota of previously root canal-treated teeth with apical periodontitis and to investigate the antibacterial effectiveness of different intracanal medicaments. Sixteen patients with post-treatment apical periodontitis were allocated into two groups according to the intracanal medicament used: calcium hydroxide (CH) and 2% chlorhexidine gluconate gel (CHX) group. Total bacterial loads, as well as the amount of Enterococcus faecalis (E. faecalis) were determined before (S1) and after (S2) chemomechanical preparation and finally, after intracanal medication (S3) by means of ddPCR. The unpaired t test was used to compare parametric. S3-total bacteria copy number of the CH group was lower than the CHX group (p < 0.05). There was no statistical difference between the CHX- and the CH groups in terms of E.faecalis copy number (p > 0.05). But in terms of total bacteria, CH is better than CHX. Consequently, CH can be used to optimise the antibacterial efficiency of chemomechanical preparation in previously root canal-treated teeth with apical periodontitis.     

The role of intracanal medication in root canal treatment

The role of intracanal medication as a root canal dressing is re-examined. In pulpectomy and some root canal treatments, where the root canal contains vital pulp tissue, it is doubtful whether a routine intracanal medicament is needed. In infected root canals, intracanal medication has been advocated for many purposes. An intracanal medicament is used to: (i) eliminate any remaining bacteria after canal instrumentation; (ii) reduce inflammation of periapical tissues and pulp remnants; (iii) render canal contents inert and neutralize tissue debris; (iv) act as a barrier against leakage from the temporary filling; (v) help to dry persistently wet canals. However, most of the indications for intracanal medicaments are questionable. Intracanal medicaments should only be used for root canal disinfection as part of controlled asepsis in infected root canals, and their role is secondary to cleaning and shaping of the root canal. Thorough canal debridement and adequate canal preparation are more pertinent, and their importance is emphasized. Bacteriological sampling may be necessary if a tooth does not respond to treatment, to help in the choice of intracanal medicament.     

Evaluation of time required for recontamination of coronally sealed canals medicated with calcium hydroxide and chlorhexidine

AIM: To determine in vitro the time required for recontamination of coronally sealed canals medicated with either calcium hydroxide (CaOH2), 2% chlorhexidine gel (CG) or with a combination of both. METHODOLOGY: Eighty intact, caries-free, premolar teeth with straight roots and mature apices were selected for the study. After biomechanical preparation of 75 teeth, they were randomly divided into nine groups according to the intracanal medicament and the coronal seal with 'Intermediate Restorative Material' (IRM) as follows: (i) 10 teeth medicated with CG, coronally unsealed; (ii) 10 teeth medicated with CaOH2, coronally unsealed; (iii) 10 teeth medicated with CaOH2 + CG, coronally unsealed; (iv) 10 teeth medicated with CG + coronal seal; (v) 10 teeth medicated with CaOH2 + coronal seal; (vi) 10 teeth medicated with CG + CaOH2 + coronal seal; (vii) 10 teeth without intracanal medicament and coronally sealed; (viii) 5 teeth without intracanal medicament and coronally unsealed, used as the positive control group (PC); (ix) 5 teeth with intact crowns used as the negative control group (NC). Glass flasks were filled with Brain Heart Infusion broth (BHI), so that only the root apex was in contact with the broth, while the crown was immersed in human saliva + BHI (3:1). The flasks were then incubated at 37 degrees C in an atmosphere of 10% CO2, and microbial growth was checked daily. RESULTS: All specimens of the PC showed contamination within 1 day of incubation, while the NC showed no evidence of broth turbidity. Recontamination was detected after an average time of 3.7 days in the unsealed canals medicated with CG, 1.8 days in the group medicated with CaOH2 and 2.6 days in the group medicated with CaOH2 + CG. When the crowns were sealed with IRM, recontamination was detected within 13.5 days in the canals medicated with CG, after 17.2 days in the group medicated with CaOH2 and after 11.9 days in the group medicated with CG + CaOH2. The group with no medication, but sealed with IRM, showed recontamination after 8.7 days. There were statistically significant differences between the teeth with or without coronal seal (P<0.05). CONCLUSION: The coronal seal delayed but did not prevent leakage of microorganisms. There was no difference between the various medicaments.     

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.     

Root canal medicaments

The ultimate goals of endodontic treatment are complete removal of bacteria, their byproducts and pulpal remnants from infected root canals and the complete seal of disinfected root canals. Intracanal medicaments have been thought an essential step in killing the bacteria in root canals; however, in modern endodontics, shaping and cleaning may be assuming greater importance than intracanal medicaments as a means of disinfecting root canals. Until recently, formocresol and its relatives were frequently used as intracanal medicaments, but it was pointed out that such bactericidal chemicals dressed in the canal distributed to the whole body from the root apex and so might induce various harmful effects including allergies. Furthermore, as these medicaments are potent carcinogenic agents, there is no indication for these chemicals in modern endodontic treatment. Today, biocompatibility and stability are essential properties for intracanal medicaments. The more modern meaning of intracanal dressing is for a blockade against coronal leakage from the gap between filling materials and cavity wall. Calcium hydroxide has been determined as suitable for use as an intracanal medicament as it is stable for long periods, harmless to the body, and bactericidal in a limited area. It also induces hard tissue formation and is effective for stopping inflammatory exudates. Single‐visit endodontics, where intracanal medicaments are not used, is generally not now contraindicated and various reports have shown that the clinical outcomes between single‐ and multiple‐ visit endodontics are similar. There is no reason to counsel against single‐visit endodontics: however, if multiple‐visit endodontics is chosen, calcium hydroxide is recommended to be used as an intracanal medicament.     

In vitro effects of calcium hydroxide and polymyxin B on endotoxins in root canals

OBJECTIVES: To evaluate the effects of intracanal medicaments on endotoxins in root canals. METHODS: Seventy-five freshly extracted maxillary incisors were used in this study. The crowns of teeth were sectioned near the CEJ in order to standardize the root length to 14 mm. The root canals were instrumented to an apical size #50 file and irrigated with 1% sodium hypochlorite solution and sterilized with 60Co gamma irradiation. Standardized suspension containing Escherichia coli endotoxin was inoculated into the 60 root canals. The specimens were randomly assigned to 5 groups (n=15), according to the intracanal medicament used: (G1) calcium hydroxide; (G2) polymyxin B; (G3) combination neomycin-polymyxin B-hydrocortisone; (G4) positive control (no intracanal medicament); (G5) negative control (no endotoxin and no intracanal medicament). After 7 days, the detoxification of endotoxin was evaluated by Limulus lysate assay and antibody production in B-lymphocytes culture. RESULTS: Groups 1, 2 and 5 presented the best results by Limulus lysate and were significantly different to groups 3 and 4 (p<0.05). Stimulation of antibodies production in cell culture by groups 1 and 6 was smaller and statistically different than groups 2, 3, 4 and 5 (p<0.05). Groups 2 and 5 induced a small increase in the antibodies production in relation to the groups 1 and 6. Groups 3 and 4 induced a significant increase of antibodies production (p<0.05). CONCLUSIONS: The calcium hydroxide and polymyxin B intracanal medicaments detoxified endotoxin in root canals and altered the properties of LPS to stimulate the antibody production by B-lymphocytes. The combination neomycin-polymyxin B-hydrocortisone did not detoxified endotoxin.     

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.     

Comparison of 2.5% Sodium Hypochlorite and 2% Chlorhexidine Gel on Oral Bacterial Lipopolysaccharide Reduction from Primarily Infected Root Canals

IntroductionThis clinical study was conducted to compare the efficacy of chemomechanical preparation with 2.5% sodium hypochlorite (NaOCl) and 2% chlorhexidine (CHX) gel on eliminating oral bacterial lipopolysaccharide (LPS) in teeth with pulp necrosis and apical periodontitis.MethodsFifty-four root canals were selected. Samples were collect before (s1) and after chemomechanical preparation (s2). Teeth were randomly divided into groups: GI, 2.5% NaOCl (n = 27) and GII, CHX gel (n = 27). Limulus amebocyte lysate assay was used to quantify endotoxins.ResultsEndotoxin was present in 100% of the samples investigated, with a median value of 272 endotoxin units (EU)/mL (GI) and of 152.46 EU/mL (GII). As a result of chemomechanical preparation, LPS content was reduced to a median value of 86 EU/mL (GI) and 85 EU/mL (GII). Higher percentage value of endotoxin reduction was found in GI (P < .05).ConclusionsThe 2.5% NaOCl and 2% CHX gel were not effective in eliminating endotoxin from the primarily infected root canals.     

Microbial Evaluation of Traumatized Teeth Treated with Triple Antibiotic Paste or Calcium Hydroxide with 2% Chlorhexidine Gel in Pulp Revascularization

Introduction

Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments.

Methods

Fifteen patients (7–17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA).

Results

Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features.

Conclusions

The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.     

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.

     

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.     

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.