The effect of a chemomechanical protocol to eliminate microorganisms from pulpectomized primary teeth: three case reports

Introduction: Description of the bacterial community before and after chemomechanical preparation (CP) with the removal of a smear layer (SL) in pulpectomized primary teeth has been little reported. Objective: These case reports describe the presence of total microorganisms and Enterococcus faecalis in root canals of primary incisors before and after CP with SL removal. Case Reports: Microbiological samples were collected from the root canals of three children (3.66±0.58 years old) with necrosis (n=2) and irreversible pulpal inflammation (n=1) in maxillary primary incisors. After teeth isolation with a rubber dam and antisepsis of the operative field, the sample collections were performed with sterile absorbent paper cones before and after the CP, which included irrigation with 2.5% sodium hypochlorite followed by 6% citric acid to remove the SL. The collected samples were analysed immediately at the end of the clinical procedures. The plates were incubated anaerobically for 48 hours at 37°C. The results were expressed as colony forming units (CFU)/mL. Results: Two of the three teeth showed total microorganisms before the CP. One incisor had no microorganisms in the initial collection. No CFU was counted in the samples collected after CP. Moreover, E. faecalis was not observed any time, either before or after the CP. Conclusions: E. faecalis was not detected in any sample, yet two of the three root canals had microorganisms before CP. In cases where microorganisms were initially found, 100% elimination was observed after the applied protocol.

Introdução: A descrição da comunidade bacteriana antes e após o preparo químico-mecânico (PQM) com remoção da smear layer (SL) em dentes decíduos pulpectomizados tem sido pouco relatada. Objetivo: Esses relatos de casos descrevem a presença de microrganismos totais e Enterococcus faecalis em canais radiculares de incisivos decíduos antes e após PQM com remoção de SL. Relatos dos Casos: Amostras microbiológicas foram coletadas do canal radicular de três crianças (3,66 ± 0,58 anos) com necrose (n = 2) e inflamação pulpar irreversível (n= 1) em incisivos decíduos superiores. Após o isolamento dos dentes com dique de borracha e antissepsia do campo operatório, as coletas das amostras foram realizadas com cones de papel absorvente estéril antes e após o PQM, que incluiu irrigação com hipoclorito de sódio 2,5% seguido de ácido cítrico 6% para retirada do SL. As amostras coletadas foram analisadas imediatamente ao final dos procedimentos clínicos. As placas foram incubadas em anaerobiose durante 48 horas a 37°C. Os resultados foram expressos em unidades formadoras de colônias (UFC)/mL. Resultados: Dois dos três dentes apresentaram microrganismos totais antes do PQM. Um incisivo não apresentava microrganismos na coleta inicial. Nenhuma UFC foi contada nas amostras coletadas após o PQM. Além disso, o E. faecalis não foi observado nenhum momento, nem antes, nem depois do PQM. Conclusão: Não foi detectado E. faecalis em nenhuma amostra, porém dois dos três canais radiculares apresentavam microrganismos antes do PQM. Nos casos em que foram encontrados microrganismos inicialmente, observou-se 100% de eliminação após o protocolo aplicado.

Antimicrobial Effect of Peptide DJK-5 Used Alone or Mixed with EDTA on Mono- and Multispecies Biofilms in Dentin Canals.

INTRODUCTION: The present study aimed to evaluate the antibacterial effect of a new peptide, DJK-5, used alone or mixed together with EDTA on mono- and multispecies biofilms in dentin canals covered by a smear layer with or without preceding sodium hypochlorite (NaOCl) irrigation. METHODS: One hundred twelve dentin blocks (224 final specimens) were prepared and divided into 56 groups, and Enterococcus faecalis or multispecies bacteria were introduced into dentinal tubules by centrifugation. After 1 week of cultivation, a uniform smear layer was created on the surface of the dentin blocks, and the samples were exposed to sterile water, 17% EDTA, 2% or 6% NaOCl, 10 µg/mL DJK-5, or a mixture of 8.5% EDTA +10 µg/mL DJK-5 or were combined treated with the solution in the following sequence: 2% or 6% NaOCl +10 µg/mL DJK-5, 2% or 6% NaOCl + 8.5% EDTA +10 µg/mL DJK-5, 2% or 6% NaOCl + 8.5% EDTA + 10 µg/mL DJK-5. Specimens without a smear layer treated by 6% NaOCl or 10 µg/mL DJK-5 served as the positive control. The irrigant exposure time was 3 or 10 minutes. The antibacterial efficacy was determined by live/dead staining and confocal laser scanning microscopy. RESULTS: The smear layer reduced the antibacterial capacity of 6% NaOCl and 10 µg/mL DJK-5. The efficacy of 2% or 6% NaOCl followed by 10 µg/mL DJK-5 was superior to 10 µg/mL DJK-5 alone (P < .05) but inferior to 2% or 6% NaOCl + 8.5% EDTA + 10 µg/mL DJK-5 and 2% or 6% NaOCl + 8.5% EDTA + 10 µg/mL DJK-5 (P < .05). The mixture of 8.5% EDTA and 10 µg/mL DJK-5 had the same disinfection effectiveness as 10 µg/mL DJK-5 used alone (P < .05). Using 2% or 6% NaOCl before EDTA + peptide always resulted in the highest killing (P < .05). CONCLUSIONS: The smear layer inhibits the disinfectant effect in dentin. Peptide DJK-5 showed a strong antibacterial effect against mono- and multispecies biofilms in dentin canals. The highest killing was measured when 6% NaOCl was followed by a mixture of EDTA and peptide DJK-5.

Efficacy of Photon-induced Photoacoustic Streaming in the Reduction of Enterococcus faecalis within the Root Canal: Different Settings and Different Sodium Hypochlorite Concentrations.

INTRODUCTION: The purpose of this study was to determine the effectiveness of laser-activated irrigation by photon-induced photoacoustic streaming (PIPS) in the reduction of Enterococcus faecalis in root canal disinfection, varying laser energy output, and sodium hypochlorite (NaOCl) concentration. For effective removal of the smear layer, the sequence and resting time of the final irrigation steps were modified compared with the standard PIPS protocol. METHODS: Eighty-six extracted single-rooted teeth were mechanically prepared, sterilized, and inoculated with E. faecalis for 4 weeks. Teeth were divided into 9 groups and treated with an Er:YAG laser using a PIPS 600/9 tip at the following parameters: 10 mJ or 20 mJ, 15 Hz, and a 50-microsecond pulse duration at 0.15 W or 0.3 W average power, respectively. Root canals were irrigated with different concentrations of NaOCl (ie, 1%, 3%, and 5% and activated using the adjusted PIPS protocol). The bacterial count was performed immediately after and 48 hours after decontamination and new incubation on an agar plate. RESULTS: A statistically significant difference in bacterial counts (P < .05) was detected in all groups before and directly after the treatment and in groups treated with 5% NaOCl 48 hours after treatment. Scanning electron microscopic imaging showed an absence of bacteria and biofilm in the scanned areas after treatment with 5% NaOCl. CONCLUSIONS: Laser-activated irrigation using 5% NaOCl and a modified PIPS protocol resulted in effective eradication of the bacterial biofilm and removal of the smear layer.

Influence of Smear Layer on the Antimicrobial Activity of a Sodium Hypochlorite/Etidronic Acid Irrigating Solution in Infected Dentin.

INTRODUCTION: The aim of this study was to evaluate the influence of the smear layer on the antimicrobial activity of a 2.5% sodium hypochlorite (NaOCl)/9% etidronic acid (HEBP) irrigating solution against bacteria growing inside dentin tubules. METHODS: Dentin tubules were infected with Enterococcus faecalis by centrifugation. After 5 days of incubation, the smear layer had formed in half of the samples, which were then treated with 2.5% NaOCl either alone or combined with 9% HEBP for 3 minutes. The percentage of dead cells in infected dentinal tubules was measured using confocal laser scanning microscopy and the live/dead technique. The smear layer on the surface of the root canal wall was also observed by scanning electron microscopy. Results of the percentage of dead cells were compared using parametric tests after subjecting data to the normalized Anscombe transformation. The level of significance was P < .05. RESULTS: In the absence of the smear layer, 2.5% NaOCl alone and combined with 9% HEBP showed high antimicrobial activity without significant differences between the 2. The smear layer reduced the antimicrobial activity of 2.5% NaOCl significantly, whereas the solution with HEBP was not affected. No dentin tubules free of the smear layer were obtained in the 2.5% NaOCl group. In the case of 2.5% NaOCl/9% HEBP, 95.40% ± 3.63% of dentin tubules were cleaned. CONCLUSIONS: The presence of the smear layer reduced the antimicrobial activity of 2.5% NaOCl. The combination of 2.5% NaOCl/9% HEBP exerted antimicrobial activity that was not reduced by the smear layer.

Effect of "apical clearing" and "apical foramen widening" on apical ramifications and bacterial load in root canals.

The purpose of the study was to determine the effect of apical clearing and apical foramen widening in reducing apical ramifications and bacterial load in the apical third of root canals. The mesio-buccal roots of 21 maxillary first molar teeth were inoculated with Enterococcus faecalis suspension using a sterile pipette. Samples were incubated at 37°C for 72 hrs and divided into 3 groups: Group A, control group (n=5), no preparation; Group B (n=8) conventional preparation alone; and Group C (n=8), apical clearing and foramen widening in addition to conventional preparation. Bacterial counts were semi-quantitatively analyzed pre- and post-preparation. Samples were demineralized with 5% nitric acid after injection of India ink. Cross sections were obtained at every 0.5 mm from the apex to 3 mm of the root using a vibratome and viewed under a stereomicroscope at 64×magnification to locate any debris or apical ramifications. The Kruskal-Wallis and Mann-Whitney U tests were used for the statistical analysis. A statistically significant difference was observed (p value 0.006) in the number of ramifications among the 3 groups. Group C had a lower average number of ramifications (1) than Group B (2.5) or A (4). The debris score was analyzed at each level (0.5-3 mm). A statistically significant difference was observed at 0.5 mm and 1 mm between Group A and C (p=0.0041) and Group B and C (p=0.0050), whereas no difference was found between Group A and B (p>0.05). These results indicate that there was less debris and fewer apical ramifications in Group C. The microbiological study revealed a lower number of colony forming units (10(2) -10(3)) in Group B or C than in Group A (>10(5)). These results suggest that apical widening and clearing facilitates removal of apical ramifications and bacterial load within root canals.

Effect of smear layer against disinfection protocols on Enterococcus faecalis-infected dentin.

INTRODUCTION: This study examined the effect of the smear layer on the antibacterial effect of different disinfecting solutions in infected dentinal tubules. METHODS: Cells of Enterococcus faecalis were forced into dentinal tubules according to a previously established protocol. After a 3-week incubation period of infected dentin blocks, a uniform smear layer was produced. Forty infected dentin specimens were prepared and subjected to 3 and 10 minutes of exposure to disinfecting solutions including sterile water, 2% and 6% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), 17% EDTA, and QMiX (Dentsply Tulsa Dental, Tulsa, OK). The following combinations were also included: 2% NaOCl + 2% CHX, 2% NaOCl + QMiX, 6% NaOCl + QMiX, and 6% NaOCl + 17% EDTA + 2% CHX. Four other dentin specimens similarly infected but with no smear layer were subjected to 3 minutes of exposure to 2% CHX and 6% NaOCl for comparison. Confocal laser scanning microscopy and viability staining were used to analyze the proportions of dead and live bacteria inside the dentin. RESULTS: In the presence of a smear layer, 10 minutes of exposure to QMiX, 2% NaOCl + QMiX, 6% NaOCl + QMiX, and 6% NaOCl + 17% EDTA + 2% CHX resulted in significantly more dead bacteria than 3 minutes of exposure to these same disinfecting solutions (P < .05). No statistically significant difference between 3 and 10 minutes was found in other groups (P > .05); 6% NaOCl + QMiX and 6% NaOCl + 17% EDTA + 2% CHX showed the strongest antibacterial effect. In the absence of a smear layer, 2% CHX and 6% NaOCl killed significantly more bacteria than they did in the presence of a smear layer (P < .05). CONCLUSIONS: The smear layer reduces the effectiveness of disinfecting agents against E. faecalis in infected dentin. Solutions containing 6% NaOCl and/or QMiX showed the highest antibacterial activity.