Antibacterial efficacy of endodontic irrigating solutions and their combinations in root canals contaminated with Enterococcus faecalis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inflammatory response to different endodontic irrigating solutions

Aim the aim of this study was to evaluate the inflammatory response to irrigating solutions injected into the peritoneal cavity of mice.Methodology Sixty mice received intra-peritoneal injections of 0.3 mL of 0.5% sodium hypochlorite, 2.0% chlorhexidine digluconate or phosphate buffered saline (PBS, control). Five animals of each group were sacrificed at 4, 24, 48 h and 7 days after the injection. Liquid from the peritoneal cavity of each animal was collected for the total and differential counting of inflammatory cells and protein leakage.Results the 0.5% sodium hypochlorite solution group had greater migration of neutrophils and mononuclear cells to the peritoneal cavity from 48 to 168 h (P < 0.05). There was a significant increase in protein leakage to the peritoneal cavity after 4 up to 48 h in the 0.5% sodium hypochlorite group compared to the control group. Protein leakage was similar in all groups at 168 h. The 2.0% chlorhexidine group had similar results to the control group at all time periods.Conclusions the 0.5% sodium hypochlorite solution induced an inflammatory response, however, the 2.0% chlorhexidine digluconate solution did not induce a significant inflammatory response.

Antibacterial effectiveness of several irrigating solutions and the Endox Plus system - an ex vivo study

Aim To compare the ex vivo antibacterial effectiveness of the Endox Plus system and sodium hypochlorite (NaOCl) in combination with BioPure MTAD (Tulsa Dental, Tulsa, OK, USA) or with EDTA in Enterococcus faecalis-contaminated root canals.Methodology After initial preparation, the root canals of 70 single-rooted human teeth were inoculated with E. faecalis (ATCC 29212) and incubated for 21 days. Specimens were divided into five groups: Endox Plus/saline; 2.5% NaOCl/MTAD; 2.5% NaOCl/EDTA; saline (positive control); negative control (root canals not prepared, nor irrigated). Samples were collected using paper points. Microbiological analysis evaluated the number of CFUs. Data were analysed by anova and Tukey tests at 0.05 significance.Results All specimens had bacterial growth after the incubation period, with similar CFU per mL counts (P > 0.05). After chemo-mechanical preparation, the number of bacteria in all groups reduced, except for the negative control. No significant differences were observed between 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA, but these groups had lower CFU counts than the other groups (P < 0.05). In the final samples, an increase in the bacterial counts was observed for Endox Plus/saline, 2.5% NaOCl/MTAD, 2.5% NaOCl/EDTA and saline (P < 0.05) with no significant differences between these groups.Conclusions This ex vivo study revealed that the Endox Plus system was associated with a reduced antibacterial effectiveness compared with conventional irrigation using 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA. All irrigation procedures allowed recovery of bacteria 7 days after treatment, demonstrating persistence of contamination within the root canal system.

In vivo microbiological evaluation of the effect of biomechanical preparation of root canals using different irrigating solutions

The aim of this study was to evaluate the antimicrobial effect of biomechanical preparation using different irrigating solutions. Seventy-eight root canals from premolars of four dogs were used. After experimental induction of periapical lesions, the root canals were prepared using the following solutions for irrigation: Group 1) 2.5% sodium hypochlorite (NaOCl); Group 2) 2% chlorhexidine (CHX); Group 3) saline solution and Group 4) control group with no biomechanical preparation. The microbiological evaluation of the root canals was performed by counting the colony forming units (CFUs) using different culture mediums. Two absorbent paper cones were used in each root canal in order to collect the microbiological samples before, and thirty days after the biomechanical preparation. The culture plates were incubated in aerobic, anaerobic and microaerophilic environment. Statistical evaluation was carried out using analysis of variance, Tukey and Student tests. The results demonstrated that there was reduction in the number of microorganisms in the NaOCl and CHX groups (p<0.05). There was greater effectiveness in the chlorhexidine group. The group that used saline solution and the control group presented an increased number of microorganisms. It can be concluded that the use of antimicrobial irrigating solutions during biomechanical preparation promotes the reduction of endodontic microbiota. However, a considerable number of microorganisms were still observed.

Antibiofilm activity of irrigating solutions associated with cetrimide. Confocal laser scanning microscopy

AimTo evaluate the antibiofilm activity of sodium hypochlorite (NaOCl) and chlorhexidine (CHX) solutions associated with cetrimide (CTR), and QMiX using confocal laser scanning microscopy.MethodologyEnterococcus faecalis (ATCC- 29212) biofilms were induced on bovine dentine blocks for 14days. The dentine blocks containing biofilm were immersed for 1min in the following solutions: 2.5% NaOCl; 2.5% NaOCl+0.2% CTR; 2% CHX; 2% CHX+0.2% CTR; 0.2% CTR; QMiX. After contact with the solutions, the dentine blocks were stained with Live/Dead((R)) BacLight for analysis of the remaining biofilm using confocal laser scanning microscope. Images were evaluated using the BioImage_L software to determine the total biovolume (m(3)), the green biovolume (live cells) (m(3)) and the percentage of substrate coverage (%). The data were subjected to nonparametric statistical test using Kruskal-Wallis and Dunn's tests at 5% significance level.ResultsAfter exposure to irrigants, the total biovolume observed for CHX, CHX+CTR, CTR, QMiX was similar to distilled water (P>0.05). NaOCl and NaOCl+CTR had the lowest total and green biovolume. The CTR and QMiX had intermediate green biovolume, with greater antibacterial activity than CHX and CHX+CTR (P<0.05). The NaOCl and NaOCl+CTR solutions were associated with microorganism removal and substrate cleaning ability.ConclusionsNaOCl and NaOCl+CTR solutions were effective on microorganism viability and were able to eliminate biofilm. The addition of cetrimide did not influence antibacterial activity.

In vitro evaluation of the action of irrigating solutions associated with intracanal medications on Escherichia coli and its endotoxin in root canals

Objective: The purpose of this study was to evaluate the efficacy of auxiliary chemical substances and intracanal medications on Escherichia coli and its endotoxin in root canals. Material and Methods: Teeth were contaminated with a suspension of E. coli for 14 days and divided into 3 groups according to the auxiliary chemical substance used: G1) 2.5% sodium hypochlorite (NaOCl); G2) 2% chlorhexidine gel (CLX); G3) pyrogen-free solution. After, these groups were subdivided according to the intracanal medication (ICM): A) Calcium hydroxide paste (Calen (R)), B) polymyxin B, and C) Calcium hydroxide paste+2% CLX gel. For the control group (G4), pyrogen-free saline solution was used without application of intracanal medication. Samples of the root canal content were collected immediately after biomechanical preparation (BMP), at 7 days after BMP, after 14 days of intracanal medication activity, and 7 days after removal of intracanal medication. The following aspects were evaluated for all collections: a) antimicrobial activity; b) quantification of endotoxin by the limulus Amebocyte lysate test (LAL). Results were analyzed by the kruskal-wallis and Dunn's tests at 5% significance level. Results: The 2.5% NaOCl and CLX were able to eliminate E. coli from root canal lumen and reduced the amount of endotoxin compared to saline. Conclusions: It was concluded that 2.5% NaOCl and CLX were effective in eliminating E. coli. Only the studied intracanal medications were to reduce the amount of endotoxin present in the root canals, regardless of the irrigant used.

Histological study of the effect of some irrigating solutions on bacterial endotoxin in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Calcium hydroxide intracanal dressing removal with different rotary instruments and irrigating solutions: A scanning electron microscopy study

This study evaluated the efficacy of 2 types of rotary instruments employed in association with sodium hypochlorite (NaOCl) or EDTA in removing calcium hydroxide (CH) residues from root canals dentin walls. Forty-two mandibular human incisors were instrumented with the ProTaper System up to F2 instrument, irrigated with 2.5% NaOCl followed by 17% EDTA and filled with a CH intracanal dressing. After 7 days, the CH dressing was removed using 4 techniques: NiTi rotary instrument size 25, 0.06 taper (K3 Endo) and irrigation with 17% EDTA (Group 1), NiTi rotary F1 instrument (ProTaper) and irrigation with 17% EDTA (Group 2), NiTi rotary instrument size 25, 0.06 taper and irrigation with 2.5% NaOCl (Group 3) and NiTi rotary F1 instrument and irrigation with 2.5% NaOCl (Group 4). Two roots without intracanal dressing were used as negative controls. Teeth were evaluated by scanning electron microscopy, in the cervical and apical canal thirds. None of the techniques removed the CH dressing completely. In the apical and cervical thirds, F1 instrument was better than instrument size 25, 0.06 taper in removing CH residues (p<0.05), regardless of the final irrigating solution. No difference was found between the irrigating solutions in the groups of F1 instrument and of instrument size 25, 0.06 taper (p>0.05). The negative controls had no CH residues on the dentin walls. In conclusion, the ProTaper F1 instrument was better than K3 Endo instrument size 25, 0.06 taper in the removal of CH intracanal medication, regardless of irrigating solution used.

Dissolution of porcine incisor pulps in sodium hypochlorite solutions of varying compositions and concentrations

Background: The solubility of dental pulp tissue in sodium hypochlorite has been extensively investigated but results have been inconsistent; due most likely to variations in experimental design, the volume and/or rate of replenishment of the solutions used and the nature of the tissues assessed. Traditionally, the sodium hypochlorite solutions used for endodontic irrigation in Australia have been either Milton or commercial bleach, with Milton being the most common. Recently, a range of Therapeutic Goods Administration (TGA) approved proprietary sodium hypochlorite solutions, which contain surfactant, has become available. Some domestic chlorine bleaches now also contain surfactants. The purpose of this study was to perform new solubility assessments, comparing Milton with new TGA approved products, Hypochlor 1% and Hypochlor 4% forte, and with a domestic bleach containing surfactant (White King). Methods: Ten randomly assigned pulp samples of porcine dental pulp of approximately equal dimensions were immersed in the above solutions, as well as representative concentrations of sodium hydroxide. Time to complete dissolution was measured and assessed statistically. Results: White King 4% showed the shortest dissolution time, closely followed by Hypochlor 4% forte. White King 1% and Hypochlor 1% each took around three times as long to completely dissolve the samples of pulp as their respective 4% concentrations, while Milton took nearly 10 times as long. The sodium hydroxide solutions showed no noticeable dissolution of the pulp samples. Conclusions: The composition and content of sodium hypochlorite solutions had a profound effect on the ability of these solutions to dissolve pulp tissue in vitro. Greater concentrations provided more rapid dissolution of tissue. One per cent solutions with added surfactant and which contained higher concentrations of sodium hydroxide were significantly more effective in dissolution of pulp tissue than Milton.

Avaliação da microbiota e da reparação apical e periapical após preparo biomecânico de canais radiculares com diferentes soluções irrigadoras, em dentes de cães com reação periapical crônica

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Endodontic 'Solutions'. Part 2:An audit comparing current practice in Belfast with UK and Republic of Ireland Dental Schools

Endodontic lubricants, irrigating solutions and medicaments help reduce the microbial load within root canals. Primary and secondary cases involve different microbes. Each'solution'or combinations thereof could play a significant role but no detailed guidelines exist on their use. An audit was undertaken to compare current practice in Belfast Dental School to the others across the UK and Republic of Ireland (ROI). This audit highlighted three main differences between Belfast and other dental schools. Many other institutions utilized other irrigants besides sodium hypochlorite (NaOCl), different intracanal medicaments, including calcium hydroxide, and higher concentrations of NaOCl. Having gathered this information, we ask, 'Is there sufficient evidence to change the endodontic regime currently used at Belfast Dental School?'. Using the findings from the literature review (Part 1), we introduce new evidence-based protocols for primary and secondary cases for use in Belfast Dental School. Clinical Relevance: In the absence of detailed clinical guidelines on the use of endodontic lubricants, irrigants and medicaments in primary and secondary cases, it is important to be aware of current practice in UK and ROI dental schools where dentists and specialists are trained.

Effect of different irrigation solutions and calcium hydroxide on bacterial LPS

Aim To evaluate the effect of biomechanical preparation with different irrigating solutions and calcium hydroxide dressing in dog root canals containing bacterial endotoxin (lipopolysaccharides; LPS).Methodology One hundred and forty premolar roots from seven dogs were filled with Escherichia coli LPS for 10 days (three roots were lost during histological processing). The following irrigating solutions were used for biomechanical preparation: 1% (group I, n = 20), 2.5% (group II, n = 19) and 5% sodium hypochlorite (group III, n = 19), 2% chlorhexidine digluconate (group IV, n = 20) and physiological saline solution (group V, n = 19). In group VI (n = 20), the LPS solution was maintained in the root canal during the entire experiment and in group VII (n = 20), after biomechanical preparation with saline solution, the root canals were filled with a calcium hydroxide dressing (Calen; control). After 60 days, the animals were sacrificed and the following parameters of periapical disease were evaluated: (a) inflammatory infiltrate, (b) periodontal ligament thickness, (c) cementum resorption and (d) bone resorption. Scores were given and data were analysed statistically with the Kruskal-Wallis and Dunn tests (P < 0.05).Results Histopathological evaluation showed that groups I-VI had more inflammatory infiltrate, greater periodontal ligament thickening and greater cementum and bone resorption (P < 0.05) compared to group VII, which received the calcium hydroxide intracanal dressing.Conclusions Biomechanical preparation with the irrigating solutions did not inactivate the effects of the endotoxin but the calcium hydroxide intracanal dressing did appear to inactivate the effects induced by the endotoxin in vivo.

Antimicrobial effect of endodontic solutions used as final irrigants on a dentine biofilm model

Aim To evaluate the residual biovolume of live bacterial cells, the mean biofilm thickness and the substratum coverage found in mixed biofilms treated with different endodontic irrigant solutions. Methodology Twenty-five bovine dentine specimens were infected intraorally using a removable orthodontic device. Five samples were used for each irrigant solution: 2% chlorhexidine, 1% sodium hypochlorite (NaOCl), 10% citric acid, 17% EDTA and distilled water. The solutions were used for 5 min. The samples were stained using the Live/Dead technique and evaluated using a confocal microscope. Differences in the amount of total biovolume (mu m3), number of surviving cells (mu m3), mean biofilm thickness (mu m) and substratum coverage (%) of the treated biofilms were determined using nonparametric statistical tests (P < 0.05). Results Similar values of biovolume total, biovolume of live subpopulations and substratum coverage were found in 2% chlorhexidine, 10% citric acid, 17% EDTA and distilled water-treated biofilms (P > 0.05). The lower values of the studied parameters were found in 1% NaOCl-treated dentine (P < 0.05) with the exception of the mean biofilm height criteria that did not reveal significant differences amongst the irrigant solutions (P > 0.05). Conclusions One per cent sodium hypochlorite was the only irrigant that had a significant effect on biofilm viability and architecture.