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Abstract Periapical lesions (PL) of endodontic origin are one of the most common pathological conditions that affect peri-radicular tissues. The main objective of this study was to evaluate the amount and species of microorganisms isolated from necrotic pulps, establish a correlation between these and the size of periapical lesions, and how the amount and species of microorganisms decreased with non-surgical root canal treatment. Twenty-seven patients with a clinical diagnosis of dental pulp necrosis and chronic periapical lesions were selected; a Cone Beam Computed Tomography (CBCT) and microbial samples of the root canal system were taken previous to a disinfection protocol, a post-instrumentation/ disinfection protocol, and a post-medication placement. Samples were processed for colony-forming unit (CFU) counting, Gram staining technique, and bacterial identification by the API-20 Strep/API-20A system. The API system identified 21 species of microorganisms in the pre-instrumentation samples, 11 species in the post-instrumentation samples, and 11 in the post-medication samples. There was a correlation coefficient of 0.598% between the initial size of the lesion and the number of bacteria, with a coefficient of determination up to 35.7%, a correlation coefficient of 0.486% and a determination coefficient of 23.6% between the size of the periapical lesion and the number of CFUs. This study contributes to the knowledge of the amount and species of microorganisms isolated and identified from necrotic pulps, establishes a correlation between the amount and species of microorganisms and the size of the periapical lesions, and shows how the decrease of microorganisms contributes to the healing of PL, corroborating the importance of an adequate disinfection protocol.

Resumen Las lesiones periapicales (LP) de origen endodóncico son la condición patológica más común que afectan los tejidos perirradiculares. El objetivo principal de este estudio es evaluar la cantidad y especie de bacterias aisladas de pulpas necróticas, correlacionar la cantidad y especies bacterianas con el tamaño de la lesión, y cómo disminuyen la cantidad y especies de microorganismos con el tratamiento de conductos. A 27 pacientes con diagnóstico de necrosis pulpar y lesión periapical crónica detectada con CBCT se les tomaron muestras microbianas del sistema de conductos antes y después del protocolo de desinfección y de la medicación intraconducto. Las muestras se procesaron para el recuento de unidades de formación de colonias (UFC), tinción de Gram e identificación mediante el sistema API-20 Strep/API-20A. Se identificaron 21 especies en las muestras pre-instrumentación, 11 en las muestras post-instrumentación y 11 en las muestras post-medicación; se observó un coeficiente de correlación del 0,598% entre el tamaño inicial de la lesión y la cantidad de bacterias, con un coeficiente de determinación hasta el 35,7%, un coeficiente de correlación del 0,486% y un coeficiente de determinación del 23,6% entre el tamaño de la lesión periapical y el número de UFCs. Este estudio contribuye al conocimiento sobre la cantidad y especies de microorganismos aislados e identificados a partir de pulpas necróticas, establece una correlación entre la cantidad y especies de microorganismos y el tamaño de las lesiones periapicales y exhibe cómo la disminución de microorganismos contribuye a la curación de LP, corroborando la importancia de un adecuado protocolo de desinfección.

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INTRODUCTION: The objective of this study was to investigate the action of photodynamic therapy on pain control after endodontic treatment in asymptomatic teeth with a primary infection, within a single visit. METHODS: Sixty (60) single-rooted teeth with pulp necrosis and periapical lesions were selected and randomly divided into two (2) groups (n = 30), according to the protocol; a control group (CG) and a group using photodynamic therapy (aPDT). The canals were instrumented with Reciproc files # 25 up to 40 along the entire length of the canal, using 2% chlorhexidine gel as the auxiliary chemical substance, followed by irrigation with sterile saline. aPDT consited of 0.005% methylene blue as photosensitizer, using AsGaAl diode laser, 660 nm wavelength, 100 mW of power and 9 J of energy, using optical fibers with 365 µm in diameter. The canals were filled with Endomethasone N cement. RESULTS: Pain intensity was assessed at 8, 12, 24, 48, 72 h and 1 week after endodontic treatment using a visual analogue scale. The level of pain was classified as none (0), mild (1-3), moderate (4-7) or severe (8-10). The data were at a significance level of 5%. There was a statistically significant difference (p<0.05) in the periods of 8, 12, 24, 48 and 72 h between the control group and the aPDT group. After 1 week, there was no statistically significant difference. CONCLUSIONS: It is concluded that photodynamic therapy had a significant effect on decreasing post-endodontic treatment pain in teeth with necrotic pulp and asymptomatic periapical lesions.

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To identify the prevalence of C. albicans in primary endodontic infections of type two diabetes mellitus (T2DM) patients and compare their clinical and radiographical characteristics with a non-diabetic control group, establishing the possible relationship between primary endodontic infection, T2DM, and C. albicans, since diabetes mellitus (DM), influences the development, course, and response to the treatment of apical periodontitis, but the presence of Candida albicans (C. albicans) has not been considered before. A total of 120 patients were selected and divided into two groups: 60 T2DM diagnosed patients and 60 non-diabetic controls. A clinical examination and radiographic analysis were performed to establish a periapical index score (PAI). Root canal samples were taken. Deoxyribonucleic acid (DNA) was extracted, and specific primers were used to identify C. albicans by polymerase chain reaction (PCR). A twofold increase in the prevalence of C. albicans in T2DM patients was observed in contrast to control patients (p = 0.0251). Sixty-five percent of T2DM patients with positive C. albicans scored a ≥ 3 PAI, while only 27% of the patients without C. albicans had a ≥ 3 PAI score (p = 0.0065). Long-term DM patients presented C. albicans more frequently (p < 0.0001). In this study, long-term T2DM patients carried C. albicans in their root canals more frequently when having a primary endodontic infection. Furthermore, this C. albicans presence seems to be related to a higher frequency of apical periodontitis.

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INTRODUCTION: This clinical study was conducted to correlate the microbiological profile and levels of endotoxins found in primary endodontic infection with the presence of clinical features and to evaluate the removal of microorganisms and endotoxins using rotary, reciprocating, and hybrid systems for biomechanical preparation. METHODS: Thirty single root canals with primary endodontic infection were evaluated with signs and symptoms and were randomly divided into 3 groups according to the instrumentation system used (n = 10) as follows: rotary Mtwo instruments (VDW, Munich, Germany) with 8 files, the reciprocating Reciproc system (VDW) with a single file, and Genius hybrid instruments with 3 files (1 rotary and 2 reciprocating files) with irrigation using 24 mL 2.5% sodium hypochlorite. Samples were collected before (S1) and after instrumentation (S2) before being submitted to microbiological culture (colony-forming units/mL) and the checkerboard DNA-DNA hybridization test. Endotoxins were quantified using the limulus amebocyte lysate assay. RESULTS: Microbiological culture showed statistical differences in the reduction of colony-forming units/mL with all systems tested (P < .05), but no statistical difference was found among the groups. The most frequently detected species were Capnocytophaga ochracea (53%) and Fusobacterium nucleatum (53%) at S1 and F. nucleatum (50%) and Leptotrichia buccalis (50%) at S2. As for the reduction of endotoxins at S2, Mtwo presented the best results (95.05%) followed by the Genius (91.85%) and Reciproc (64.68%) groups, but no statistical difference was found among the groups. Previous pain, tenderness to percussion, and presence of a sinus tract were associated with specific microorganisms (P < .05). CONCLUSIONS: Signs and symptoms were correlated with microorganisms. Endodontic treatment was effective in reducing bacteria and endotoxins but was not capable of completely removing them from the root canal.

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Os objetivos do presente estudo foram: a) Verificar a carga microbiana e quantificar endotoxinas presentes nos canais radiculares de dentes com infecção endodôntica primária; b) Comparar a efetividade antibacteriana e sobre endotoxinas do preparo biomecânico com dois métodos de irrigação final: com ou sem irrigação ultrassônica passiva; c) Correlacionar sinais e sintomas com a microbiota presente e complexos bacterianos; d) Relacionar níveis de endotoxinas e carga microbiana com sinais e sintomas clinicos. Foram selecionados para o estudo 20 dentes uniradiculares, que apresentavam infecção endodôntica primária e lesão periapical, sendo estes submetidos a TCFC. Os canais foram preparados com sistema reciprocante e irrigação com NaOCl 2.5%, seguido de dois protocolos de irrigação final: com e sem uso de ultrassom. Todos os canais foram preenchidos com MIC com hidróxido de cálcio + soro fisiológico e após 14 dias foram obturados com cones de guta-percha e cimento AH Plus. Foram realizadas coletas do conteúdo do canal, logo após a abertura coronária (1ª Coleta); após PBM (2ª Coleta) e após limpeza final dos canais com NaOCl seguido de EDTA por 5 min (grupo sem PUI); ou irrigação com NaOCl + PUI e EDTA + PUI (3ª Coleta) (grupo com PUI) . Foram avaliados sinais/sintomas clínicos previo ao tratamento.O conteúdo das coletas foi analisado a partir de sondas de DNA pelo método de hibridização DNA-DNA checkerboard e foi realizado teste de avaliação da atividade antimicrobiana por cultura microbiológica. Alem disso foi realizado quantificação de endotoxinas pelo Lisado de Amebócitos de Limulus (LAL). E a volumetria dos canais foi feita através da TCFC utilizando o software Nemotec®. Os resultados foram submetidos a análise estatístico pelos testes Mann Whitney e teste ANOVA a dois fatores de medidas repetidas (5%). Valor de p < 0.05 foi considerado estatisticamente significante. Houve associação entre dor à percussão e complexos laranja principal e laranja periférico. Fístula foi associada ao complexo laranja principal e laranja periférico. Foi observada redução de bactérias após o tratamento endodôntico. Ao utilizar o método de hibridização DNA-DNA checkerboard, as bactérias mais prevalentes foram S. constellatus, E faecalis, F. nucleatum, P. gingivalis, P. melaninogenica e S. intermedius na primeira coleta. Os métodos de irrigação diminuíram número de espécies bacterianas comparado à coleta inicial. A irrigação ultrassônica passiva apresentou diferença estatística significativa sobre bactérias anaeróbias. PUI não apresento diferença significativa sobre bactérias aeróbias nem endotoxinas comparado com irrigação convencional. Concluiu-se que todos os canais apresentaram micro-organismos sendo que anaeróbios estiveram presentes em grande quantidade. Após preparo biomecânico do canal radicular e irrigação com PUI, houve redução significante da carga de micro-organismos anaeróbios. Os micro-organismos mais resistentes ao preparo biomecânico com e sem PUI foram S. constellatus , E. faecalis, S. intermedius e L.bucalis. Houve associação positiva entre sinais e sintomas clínicos com complexo laranja e com micro-organismos. Os níveis de endotoxinas diminuíram após o preparo biomecânico não foram inferenciados por PUI a irrigação convencional. (AU)

The objective of this study were: a) to check the microbial load and to quantify endotoxin present in the root canals of teeth with primary endodontic infection; b) to compare the effectiveness of bacteria and endotoxin reduction of the biomechanical preparation with final irrigation methods: with or without passive ultrasonic irrigation (PUI); c) to correlate signs and symptoms with the present microbiota and bacterial complexes; d) to relate endotoxin and microbial load levels with signs and symptoms before biomechanical preparation. Twenty single-root teeth were selected for the study, presenting primary endodontic infections and periapical lesion, which underwent CBCT. The canals were prepared with reciprocating system and irrigation with 2.5% NaOCl, followed by two final irrigation protocols: with and without ultrasound. All canals were filled with MIC with calcium hydroxide + saline and after 14 days were filled with gutta percha and AH Plus. Samples from the content of canal were harvested, after coronal opening (1st Sample); after PBM (2nd Sample) and after final cleaning of the canals with NaOCl followed by EDTA for 5 min (group without PUI); or irrigation with NaOCl + PUI and EDTA + PUI (3rd Sample) (group with PUI). The contents of the samples were analyzed from DNA probes by the DNA-DNA checkerboard hybridization method and a test was performed to evaluate the antimicrobial activity by microbiological culture. In addition, quantification of endotoxins by Limulus Amebocyte Lysate (LAL) was performed. The canal volumetry was assessed through the CBCT using Nemotec® software. The results were submitted to statistical analysis by Mann Whitney and repeated measures two-way ANOVA (5%). Value of p <0.05 was considered statistically significant. There was an association between pain at percussion and main orange and peripheral orange complexes. Fistula was associated with the main orange and peripheral orange complex. A reduction of bacteria was observed after endodontic treatment. When using the DNA-DNA checkerboard hybridization method, the most prevalent bacterias were S. constellatus, E faecalis, F. nucleatum, P. gingivalis, P. melaninogenica and S. intermedius in the first collection. Irrigation methods decreased number of bacterial species compared to initial collection. The passive ultrasonic irrigation presents statistically significant difference on anaerobic bacteria. PUI did not present significant difference on aerobic bacteria or endotoxins compared to conventional irrigation. In conclusion, all canals had microorganisms, and anaerobes were present in large quantities. The most frequent microorganisms were S. constellatus, E. faecalis, F. nucleatum SP, P. gingivalis e S. intermedius. After biomechanical preparation of root canal and irrigation with PUI, there was a significant reduction of the anaerobic microorganism load. The microorganisms most resistant to biomechanical preparation with and without PUI were S. constellatus , E. faecalis, S. intermedius e L. bucalis. There was a positive association between clinical signs and symtoms with orange complex and with microorganisms. Endotoxi levels decreased after biomechanical preparation, while not influenced by PUI in comparison to conventional irrigation(AU)

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