Effect of passive ultrasonic activation on microorganisms in primary root canal infection: a randomized clinical trial.

OBJECTIVE: This clinical study sought to evaluate the effectiveness of passive ultrasonic activation (PUA) in eliminating microorganisms in primary endodontic infection (PEI) after instrumentation of root canals using microbiological culture and checkerboard DNA-DNA hybridization. METHODOLOGY: Twenty root canals with PEI and apical periodontitis were selected. The root canals were instrumented and then randomly divided into 2 groups, according to the irrigation method: PUA and conventional needle irrigation (CNI). Microbiological samples were collected before instrumentation (S1), after instrumentation (S2) and after irrigation with 17% EDTA (S3). The samples were subjected to anaerobic culture technique and checkerboard DNA-DNA hybridization analysis. RESULTS: A statistically significant difference was found between CNI (23.56%) and PUA (98.37%) regarding the median percentage values for culturable bacteria reduction (p<0.05). In the initial samples, the most frequently detected species was S. constellatus (50%), and after root canal treatment was E. faecalis (50%). CONCLUSION: Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, no statistical difference in the total microbial load between PUA and CNI groups was detected. The number of cultivable anaerobic bacteria reduced significantly using PUA, and the bacterial composition and number of bacterial species after using either CNI or PUA was similar.

The influence of adrenergic blockade in rats with apical periodontitis under chronic stress conditions.

OBJECTIVE: To investigate the influence of chronic stress and adrenergic blockade in a rat model of apical periodontitis. METHODS: Thirty-two Wistar rats were submitted to an animal model of periapical lesion and randomly divided into 4 groups (n = 8): no stress (NS); stress + saline solution (SS); stress + ß-adrenergic blocker (Sß); stress + α-adrenergic blocker (Sα). The SS, Sß and Sα groups were submitted to an animal model of chronic stress for 28 days and received daily injections of saline solution, propranolol (ß adrenergic blocker) and phentolamine (α adrenergic blocker), respectively. After 28 days the animals were euthanized and the following analyses were carried out: a) serum corticosterone levels through Radioimmunoassay; b) measurement of serum levels of IL-1B, IL-6, IL-10 and IL-17 by enzyme-linked immunosorbent assay (ELISA); c) volume of periapical bone resorption by micro-computed tomography; d) histomorphometric analysis by staining with hematoxylin and eosin; e) expression of ß-AR, α-AR, receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) by immunohistochemistry; f) tartrate-resistant acid phosphatase (TRAP) staining; g) ex-vivo cytokine release followed by the stimulation with LPS in superfusion system, by ELISA. RESULTS: SS group displayed significantly higher corticosterone levels than NS group (non-stressed). Higher IL-1ß serum level was observed in the NS group (p < .05); compared to all stressed groups. Other cytokines were present in similar amounts in the serum of all groups. All groups presented similar periapical lesions. All groups presented moderate inflammatory infiltrate, without statistically significant differences between them. No differences were observed regarding ß-AR, α-AR, Rank-L and OPG expression. The number of TRAP-positive cells was significantly decreased in the groups that received daily injections of adrenergic blockers. The IL-1ß release followed LPS stimulation was significantly suppressed when the superfusion media contained propranolol (p < .05). Perfusion containing phentolamine induced a greater release of IL-10. TGF-ß was significantly suppressed by phentolamine perfusion in the NS group (p < .05). CONCLUSIONS: Chronic stress can significantly change the inflammatory cytokines release. Rank-L/OPG system and periapical lesion volume were not affected following the current method applied. The administration of adrenergic blockers was not able to modulate the inflammatory response but presented effectivity in reducing the number of osteoclasts in the periapical region.

Effect of passive ultrasonic activation on microorganisms in primary root canal infection: a randomized clinical trial

Abstract Objective: This clinical study sought to evaluate the effectiveness of passive ultrasonic activation (PUA) in eliminating microorganisms in primary endodontic infection (PEI) after instrumentation of root canals using microbiological culture and checkerboard DNA-DNA hybridization. Methodology: Twenty root canals with PEI and apical periodontitis were selected. The root canals were instrumented and then randomly divided into 2 groups, according to the irrigation method: PUA and conventional needle irrigation (CNI). Microbiological samples were collected before instrumentation (S1), after instrumentation (S2) and after irrigation with 17% EDTA (S3). The samples were subjected to anaerobic culture technique and checkerboard DNA-DNA hybridization analysis. Results: A statistically significant difference was found between CNI (23.56%) and PUA (98.37%) regarding the median percentage values for culturable bacteria reduction (p<0.05). In the initial samples, the most frequently detected species was S. constellatus (50%), and after root canal treatment was E. faecalis (50%). Conclusion: Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, no statistical difference in the total microbial load between PUA and CNI groups was detected. The number of cultivable anaerobic bacteria reduced significantly using PUA, and the bacterial composition and number of bacterial species after using either CNI or PUA was similar.

Microbial Profile and Endotoxin Levels in Primary Periodontal Lesions with Secondary Endodontic Involvement.

This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Microbial profile and endotoxin levels in primary periodontal lesions with secondary endodontic involvement

Abstract This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Resumo Este estudo foi realizado para investigar o perfil microbiano e os níveis de endotoxina de lesões endoperiodontais de origem periodontal. Amostras periodontais e endodônticas foram obtidas de bolsas periodontais e canais radiculares necróticos de 10 dentes com lesões endoperiodontais. A investigação de 40 espécies bacterianas diferentes foram determinadas em cada amostra endodôntica e periodontal usando o método de hibridização de DNA-DNA (checkerboard) e o ensaio cinético cromogênico LAL foi usado para quantificação de endotoxinas. O teste exato de Fisher correlacionou as espécies bacterianas com a microbiota endodôntica ou periodontal. Os níveis de endotoxina (EU/mL) encontrados nas amostras do canal radicular e na bolsa periodontal foram comparados pelo teste de Wilcoxon (p<0,05). Bactérias e unidades de LPS foram encontradas em 100% das amostras endodônticas e periodontais. As espécies E. faecium, P. acnes, G. morbillorum, C. sputigena e L. buccalis foram fortemente correlacionadas com a microbiota endodôntica e P. nigrescens com a microbiota periodontal. P. intermedia, P. endodontalis e V. parvula foram mais prevalentes em ambas microbiotas endodôntica e periodontal. Os níveis de endotoxina na bolsa periodontal (89600 EU/mL) foram significativamente maiores do que no canal radicular (2310 EU/mL). Concluiu-se que a microbiota presente nos tecidos periodontal e endodôntico é semelhante, com maior prevalência de espécies do complexo laranja e maior nível de endotoxina nas bolsas periodontais.

Correlation Between Volume of Root Canal, Cultivable Bacteria, Bacterial Complexes and Endotoxins in Primary Infection.

This clinical study was conducted to correlate the levels of endotoxins and culturable bacteria found in primary endodontic infection (PEI) with the volume of root canal determined by using Cone Beam Computed Tomography (CBCT); and to evaluate the bacterial diversity correlating with clinical features. Twenty patients with PEI were selected and clinical features were recorded. The volume (mm3) of root canal was determined by CBCT analysis. Root canal samples were analyzed by using kinetic LAL-assay test to determine the levels of endotoxins and anaerobic technique to determine the bacterial count (CFU/mL). DNA was extracted from all samples to determine bacterial diversity and quantified by using Checkerboard-DNA-DNA- Hybridization. Culturable bacteria and endotoxins were detected in 100% of the root canal samples. Linear regression analysis revealed a correlation between root canal volume and presence of anaerobic bacteria (p<0.05). Positive correlations were found between bacteria species and presence of different clinical features (p<0.05). After grouping the bacteria species into bacterial complexes, positive associations were found between green, orange and red complexes with presence of sinus tract (p<0.05). This clinical study revealed that larger root canals hold higher levels of culturable bacteria in PEI. Thus, the interaction of different virulent bacteria species in complexes seems to play an important role in the development of clinical features.

Effectiveness in the Removal of Endotoxins and Microbiological Profile in Primary Endodontic Infections Using 3 Different Instrumentation Systems: A Randomized Clinical Study.

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.

Limewater and Polymyxin B Associated with NaOCl for Endotoxin Detoxification in Root Canal with Necrotic Pulp.

This clinical study investigated the effects of endodontic treatment by using different irrigants (limewater + NaOCl and polymyxin B + NaOCl) and intracanal medication on endotoxins in teeth with primary endodontic infection and radiographically visible apical periodontitis. Thirty-three teeth with necrotic pulp and periapical lesions from different patients were selected for this study. Samples were collected after the coronal opening (S1) and after instrumentation (S2). Root canals were divided in 3 groups (n = 11) according to the irrigant combination used: NaOCl + LW: 2.5% NaOCl + calcium hydroxide solution (0.14%, limewater); NaOCl + PmB: 2.5% NaOCl + 10.000 UI/mL polymyxin B; 2.5% NaOCl (control). The third sampling (S3) was performed after ethylenediaminetetraacetic acid and the fourth (S4) after samples got 14 days with intracanal medication with 2% chlorhexidine gel + calcium hydroxide. Endotoxins (lipopolysaccharide) were quantified by chromogenic Limulus amebocyte lysate (LAL). Endotoxins were detected in all root canals after the coronal opening (S1). NaOCl + PmB group presented the greatest endotoxin reduction after instrumentation (76.17%), similar to NaOCl + LW group (67.64%, p<0.05) and different from NaOCl group (42.17%, p<0.05). After intracanal medication period (S4), there was significant increase of endotoxins neutralization. It was concluded that NaOCl + PmB promoted the greatest reduction of endotoxin levels, followed by NaOCl + LW. Intracanal medications had no significant complementary role in the reduction of endotoxins at the end of the treatment.

Limewater and Polymyxin B Associated with NaOCl for Endotoxin Detoxification in Root Canal with Necrotic Pulp

Abstract This clinical study investigated the effects of endodontic treatment by using different irrigants (limewater + NaOCl and polymyxin B + NaOCl) and intracanal medication on endotoxins in teeth with primary endodontic infection and radiographically visible apical periodontitis. Thirty-three teeth with necrotic pulp and periapical lesions from different patients were selected for this study. Samples were collected after the coronal opening (S1) and after instrumentation (S2). Root canals were divided in 3 groups (n = 11) according to the irrigant combination used: NaOCl + LW: 2.5% NaOCl + calcium hydroxide solution (0.14%, limewater); NaOCl + PmB: 2.5% NaOCl + 10.000 UI/mL polymyxin B; 2.5% NaOCl (control). The third sampling (S3) was performed after ethylenediaminetetraacetic acid and the fourth (S4) after samples got 14 days with intracanal medication with 2% chlorhexidine gel + calcium hydroxide. Endotoxins (lipopolysaccharide) were quantified by chromogenic Limulus amebocyte lysate (LAL). Endotoxins were detected in all root canals after the coronal opening (S1). NaOCl + PmB group presented the greatest endotoxin reduction after instrumentation (76.17%), similar to NaOCl + LW group (67.64%, p<0.05) and different from NaOCl group (42.17%, p<0.05). After intracanal medication period (S4), there was significant increase of endotoxins neutralization. It was concluded that NaOCl + PmB promoted the greatest reduction of endotoxin levels, followed by NaOCl + LW. Intracanal medications had no significant complementary role in the reduction of endotoxins at the end of the treatment

Resumo Este estudo clínico investigou os efeitos do tratamento endodôntico com uso de diferentes irrigantes (NaOCl + água de cal e NaOCl + polimixina B) e medicação intracanal sobre endotoxinas em dentes com infecção endodôntica primária e presença de lesão periapical visível radiograficamente. Foram selecionados para o estudo trinta e três dentes de pacientes que apresentavam necrose pulpar e presença de lesão periapical. As amostras foram coletadas após a abertura coronária (S1) e após a instrumentação (S2). Os canais radiculares foram divididos em 3 grupos (n = 11) de acordo com a combinação de irrigantes utilizada: NaOCl + LW:- hipoclorito de sódio 2,5% + solução de hidróxido de cálcio (água de cal 0,14%); NaOCl + PmB: hipoclorito de sódio a 2,5% + polimixina B 10.000 UI/mL; NaOCl (controle): hipoclorito de sódio a 2,5%. A terceira coleta (S3) foi realizada após aplicação do ácido etilenodiamino tetra acético (EDTA) e a quarta coleta (S4) após 14 dias de medicação intracanal de hidróxido de cálcio + clorexidina gel 2%. Endotoxinas (lipopolissacarídeos) foram quantificadas pelo ensaio cromogênico do lisado de amebócitos de Limulus (LAL). Endotoxinas foram detectadas em todos os canais radiculares após abertura coronária (S1). Grupo NaOCl + PmB apresentou a maior redução de endotoxinas após a instrumentação (76,17%), sendo similar ao grupo NaOCl + LW (67,64%, P >.05) e diferente do grupo NaOCl (42,17%, P <.05). Após o período de medicação intracanal, houve aumento significativo da neutralização de endotoxinas. Concluiu-se que NaOCl + PmB promoveu a maior redução dos níveis de endotoxinas, seguido de NaOCl + LW. A medicação intracanal não teve um papel complementar significativo na redução de endotoxinas no final do tratamento.

Investigation of Bacterial Contents From Persistent Endodontic Infection and Evaluation of Their Inflammatory Potential.

This clinical study investigated and quantified cultivable bacteria and their levels of endotoxins in persistent endodontic infection, determining their antigenicity against macrophages and fibroblast cells by IL-1ß and TNF-α secretion and evaluating their relationship with clinical and radiographic features. Samples from the root canals were obtained after root filling removal. Culture techniques were used to determine the bacterial count and the endotoxins were determined by LAL-assay. PCR analysis (16S rDNA) was used for bacterial detection. Raw 264.5 macrophages and V79 fibroblast were stimulated with endodontic contents. ELISA assay measured the amounts of IL-1ß/TNF-?#61537; secretion. Bacteria and endotoxin medians were 1.24x105 CFU/mL and 9.62 EU/mL, respectively. Porphyromonas endodontalis was the most frequently detected species. Higher levels of endotoxins were found in teeth with pain on palpation (23.56 EU/mL) rather than in its absence (8.21 EU/mL). Larger areas of bone destruction were related to higher levels of endotoxins and IL-1ß and TNF-α secretion. The study findings revealed the presence of Gram-negative bacteria species in persistent endodontic infection, with their endotoxins related to both severity of bone destruction and development of symptomatology. Moreover, larger areas of bone destruction were related to higher levels of IL-1ß and TNF-α secreted by macrophages and fibroblast cells.

Investigation of Bacterial Contents From Persistent Endodontic Infection and Evaluation of Their Inflammatory Potential

Abstract This clinical study investigated and quantified cultivable bacteria and their levels of endotoxins in persistent endodontic infection, determining their antigenicity against macrophages and fibroblast cells by IL-1β and TNF-α secretion and evaluating their relationship with clinical and radiographic features. Samples from the root canals were obtained after root filling removal. Culture techniques were used to determine the bacterial count and the endotoxins were determined by LAL-assay. PCR analysis (16S rDNA) was used for bacterial detection. Raw 264.5 macrophages and V79 fibroblast were stimulated with endodontic contents. ELISA assay measured the amounts of IL-1ß/TNF-?#61537; secretion. Bacteria and endotoxin medians were 1.24x105 CFU/mL and 9.62 EU/mL, respectively. Porphyromonas endodontalis was the most frequently detected species. Higher levels of endotoxins were found in teeth with pain on palpation (23.56 EU/mL) rather than in its absence (8.21 EU/mL). Larger areas of bone destruction were related to higher levels of endotoxins and IL-1β and TNF-α secretion. The study findings revealed the presence of Gram-negative bacteria species in persistent endodontic infection, with their endotoxins related to both severity of bone destruction and development of symptomatology. Moreover, larger areas of bone destruction were related to higher levels of IL-1β and TNF-α secreted by macrophages and fibroblast cells.

Resumo Este estudo clínico investigou e quantificou bactérias cultiváveis e seus níveis de endotoxinas na infecção endodôntica persistente, determinando a sua antigenicidade contra macrófagos e células de fibroblastos através de IL-1β e TNF-α; e avaliando sua relação com características clínicas e radiográficas. As amostras dos canais radiculares foram obtidas após a desobturação. Técnicas de cultura foram utilizadas para determinar a contagem de bactérias e a quantificação de endotoxinas foram determinadas por ensaio de LAL. Análise por PCR (16S rDNA) foi utilizada para a detecção bacteriana. Células 264,5 macrófagos e fibroblastos V79 foram estimuladas com conteúdo endodôntico. IL-1β e TNF-α produzidas pelas células avaliadas foram medidas por ensaio de ELISA. As medianas de bactérias e endotoxinas foram 1,24x105 UFC/mL e 9,62 EU/mL, respectivamente. Porphyromonas endodontalis foi a espécie mais frequentemente detectada. Níveis mais elevados de endotoxinas foram encontrados em dentes com dor à palpação (23,56 EU/mL) quando comparado a sua ausência (8,21 EU/mL). Maiores áreas de destruição óssea foram relacionados com níveis mais elevados de endotoxinas e IL-1β e TNF-α. O estudo revelou presença de espécies de bactérias Gram-negativas em infecção endodôntica persistente, com níveis elevados de endotoxinas relacionados a maior destruição óssea periapical e presença de sintomatologia. Além disso, grandes áreas de destruição óssea foram relacionados com níveis mais elevados de IL-1β e TNF-α secretadas por macrófagos e fibroblastos.

Comparison of Different Irrigants in the Removal of Endotoxins and Cultivable Microorganisms from Infected Root Canals.

This study was conducted to compare the effectiveness of different irrigants used to remove endotoxins and cultivable microorganisms during endodontic therapy. Forty root canals were contaminated and divided into groups according to the irrigant: 2% NaOCl + surfactant, 2% CHX, 2.5% NaOCl, and pyrogen-free saline solution (control). Samples were collected after root canal contamination (S1), after instrumentation (S2), and 7 days after instrumentation (S3). Microorganisms and endotoxins were recovered from 100% of the contaminated root canals (S1). At S2, 2% NaOCl + surfactant, 2% CHX, and 2.5% NaOCl were able to completely eliminate cultivable microorganisms. At S3, both 2% CHX and 2.5% NaOCl were effective in preventing C. albicans and E. coli regrowth, but E. faecalis was still detected. No microorganism species was recovered from root canals instrumented with 2% NaOCl + surfactant. At S2, a higher percentage value of endotoxin reduction was found for 2% NaOCl + surfactant (99.3%) compared to 2% CHX (98.9%) and 2.5% NaOCl (97.18%) (p < 0.05). Moreover, at S3, 2% NaOCl + surfactant (100%) was the most effective irrigant against endotoxins. All irrigants tested were effective in reducing microorganisms and endotoxins from root canals. Moreover, 2% NaOCl + surfactant was the most effective irrigant against endotoxins and regrowth of microorganisms.

In vitro antimicrobial and anti-endotoxin action of Zingiber Officinale as auxiliary chemical and medicament combined to calcium hydroxide and chlorhexidine.

OBJECTIVE: This study was conducted in vitro to compare the effectiveness of Zingiber Officinale as an auxiliary chemical substance followed by placement of different intra-canal medication in removing endotoxins and cultivable micro-organisms from infected root canals. MATERIALS AND METHODS: Seventy-two root canals were contaminated with Enterococcus faecalis, Candida albicans and Escherichia coli for 28 days. After, the teeth were instrumented using Zingiber Officinale and divided into six groups according to the intra-canal medication: chlorhexidine gel; calcium hydroxide + chlorhexidine gel; glycolic ginger extract; calcium hydroxide + glycolic ginger extract; calcium hydroxide + saline solution and saline solution (control). Sample collections were performed after root canal contamination (Baseline; S1), after instrumentation (S2), 7 days after instrumentation (S3), after 14 days with intra-canal medication (S4) and 7 days after removal of intra-canal medication (S5). The results were analyzed by the Kruskal-Wallis and Dunn tests. RESULTS: It was observed that in S2 and S3 there was significant reduction of the micro-organisms and the quantity of endotoxins after instrumentation. In samples S4 and S5 there was complete elimination of micro-organisms and significant reduction of endotoxins. CONCLUSION: It was concluded that Zingiber Officinale as an auxiliary chemical substance was effective on the micro-organisms tested, yet was unable to eliminate the endotoxins. Similarly, the intra-canal medication were effective on micro-organisms, yet did not completely eliminate the endotoxins.

Quantificação de endotoxinas e sua relação com sinais /sintomas e volumetria da lesão periapical e do canal radicular em dentes com infecção endodôntica primária / Quantification of endotoxins and its relation with signals/symptoms and volume of periapical lesion and volume of the lumen of the canal in teeth with primary endodontic infection

Os objetivos do presente estudo foram: a) quantificar endotoxinas e cargamicrobiana nas infecções endodônticas primárias, antes e após o preparobiomecânico (PBM) e uso de diferentes medicações intracanais (MIC); b)relacionar níveis de endotoxinas e microrganismos cultiváveis com avolumetria da lesão periapical (VLP), através do uso de tomografiascomputadorizadas de feixe cônico (TCFC), verificando possíveis relaçõescom sinais e sintomas clínicos; c) relacionar os mesmos níveis com avolumetria dos canais radiculares (VCR); d) Comparar medidas das lesões utilizando radiografias periapicais (RP) e tomografias computadorizadas de feixe cônico (TCFC). Foram selecionados para o estudo trinta dentes com necrose pulpar e lesão periapical que foramsubmetidos a TCFC. Após abertura coronária, foi realizada coleta inicialpara verificação da presença de infecção nos canais radiculares. Após, procedeu-se o tratamento endodôntico utilizando solução de NaOCl 2,5% e divididos em 3 grupos de acordo com a MIC: Ca(OH)2 - hidróxido de cálcio P.A. + solução salina fisiológica; Ca(OH)2 + GEN - hidróxido de cálcio P.A. + extrato glicólico de gengibre 20% e Ca(OH)2 + CLX -hidróxido de cálcio P.A. + clorexidina gel 2%. Foram realizadas coletas docanal radicular após o PBM e após 14 dias de ação da MIC. Para todasas coletas foram realizados testes de atividade antimicrobiana por culturamicrobiológica e análise da quantificação de endotoxinas pelo lisado deamebócitos de Limulus. Foi realizada a VLP e a VCR através da TCFC; ecomparadas medidas de segmentação 2D das lesões periapicais das RP e das TCFC. Todos os dados foram analisados estatisticamente. Os resultados mostraram: a) presença de microrganismos na coleta inicialvariando de 0 – 8,16 x 106 UFC/mL e níveis de endotoxinas de 1,75 – 149EU/mL; havendo redução significante de seus níveis após PBM e uso de MIC; b) Houve correlação positiva entre a VLP com os níveis de endotoxinas e presença de dor prévia, além de forte...

The aims of the present study were: a) quantify endotoxins and bacterial CFU counts in primary endodontic infections before and after biomechanical preparation (BP) and after root canal medication (RCM); b) to correlate the levels of endotoxins and bacterial CFU counts with the volume of periapical bone destruction (VPBD) determined by Cone Beam Computed Tomography (CBCT) analysis as well as with the development of clinical signs and symptoms; c) to correlate the levels of endotoxins and bacterial CFU counts with the volume of the lumen of the canal (VLC); d) to correlate the size of periapical lesions determined by periapical radiographs images with the CBCT images. Thirty teeth with pulp necrosis and apical periodontitis were selected and submitted to CBCT analysis.After access to pulp cavity, a first root canal sampling was performed inorder to determine the presence of root canal infection. Afterwards, BP was performed using 2.5% NaOCl followed by the placement of RCM according to the selection group: Calcium Hydroxide [Ca(OH)2] – Ca(OH)2 + Saline Solution (SSL); Ca(OH)2 + Ginger – Ca(OH)2 + 20% glycolic Ginger extract ; Ca(OH)2 + Chlorhexidine (CHX). A second collect were made after the BP and then the third after 14-days of RCM. Cultureanalysis was performed in order to determine antimicrobial activity. Thelevels of endotoxins were determined by the Limulus Ameboyte Lysate assay. The VPBD and the VLC were determined by CBCT-analysis and compared to periapical radiographs. Data were typed on a spreadsheet and statistical analyzed. At the baseline samples, CFU counts and endotoxins levels ranged from 0 - 8.16 CFU/mL and 1.75 -149 EU/mL, respectively. Both contents were reduced after BP and after RCM. Positive correlations were found between the levels of endotoxins and VPBD as well as with previous episode of pain. Thereby, a positive correlation was found between the bacterial CFU counts with the endotoxins contents. ...

Quantificação de endotoxinas e sua relação com sinais /sintomas e volumetria da lesão periapical e do canal radicular em dentes com infecção endodôntica primária / Quantification of endotoxins and its relation with signals/symptoms and volume of periapical lesion and volume of the lumen of the canal in teeth with primary endodontic infection

Os objetivos do presente estudo foram: a) quantificar endotoxinas e cargamicrobiana nas infecções endodônticas primárias, antes e após o preparobiomecânico (PBM) e uso de diferentes medicações intracanais (MIC); b)relacionar níveis de endotoxinas e microrganismos cultiváveis com avolumetria da lesão periapical (VLP), através do uso de tomografiascomputadorizadas de feixe cônico (TCFC), verificando possíveis relaçõescom sinais e sintomas clínicos; c) relacionar os mesmos níveis com avolumetria dos canais radiculares (VCR); d) Comparar medidas das lesões utilizando radiografias periapicais (RP) e tomografias computadorizadas de feixe cônico (TCFC). Foram selecionados para o estudo trinta dentes com necrose pulpar e lesão periapical que foramsubmetidos a TCFC. Após abertura coronária, foi realizada coleta inicialpara verificação da presença de infecção nos canais radiculares. Após, procedeu-se o tratamento endodôntico utilizando solução de NaOCl 2,5% e divididos em 3 grupos de acordo com a MIC: Ca(OH)2 - hidróxido de cálcio P.A. + solução salina fisiológica; Ca(OH)2 + GEN - hidróxido de cálcio P.A. + extrato glicólico de gengibre 20% e Ca(OH)2 + CLX -hidróxido de cálcio P.A. + clorexidina gel 2%. Foram realizadas coletas docanal radicular após o PBM e após 14 dias de ação da MIC. Para todasas coletas foram realizados testes de atividade antimicrobiana por culturamicrobiológica e análise da quantificação de endotoxinas pelo lisado deamebócitos de Limulus. Foi realizada a VLP e a VCR através da TCFC; ecomparadas medidas de segmentação 2D das lesões periapicais das RP e das TCFC. Todos os dados foram analisados estatisticamente. Os resultados mostraram: a) presença de microrganismos na coleta inicialvariando de 0 – 8,16 x 106 UFC/mL e níveis de endotoxinas de 1,75 – 149EU/mL; havendo redução significante de seus níveis após PBM e uso de MIC; b) Houve correlação positiva entre a VLP com os níveis de endotoxinas e presença de dor prévia, além de forte...

The aims of the present study were: a) quantify endotoxins and bacterial CFU counts in primary endodontic infections before and after biomechanical preparation (BP) and after root canal medication (RCM); b) to correlate the levels of endotoxins and bacterial CFU counts with the volume of periapical bone destruction (VPBD) determined by Cone Beam Computed Tomography (CBCT) analysis as well as with the development of clinical signs and symptoms; c) to correlate the levels of endotoxins and bacterial CFU counts with the volume of the lumen of the canal (VLC); d) to correlate the size of periapical lesions determined by periapical radiographs images with the CBCT images. Thirty teeth with pulp necrosis and apical periodontitis were selected and submitted to CBCT analysis.After access to pulp cavity, a first root canal sampling was performed inorder to determine the presence of root canal infection. Afterwards, BP was performed using 2.5% NaOCl followed by the placement of RCM according to the selection group: Calcium Hydroxide [Ca(OH)2] – Ca(OH)2 + Saline Solution (SSL); Ca(OH)2 + Ginger – Ca(OH)2 + 20% glycolic Ginger extract ; Ca(OH)2 + Chlorhexidine (CHX). A second collect were made after the BP and then the third after 14-days of RCM. Cultureanalysis was performed in order to determine antimicrobial activity. Thelevels of endotoxins were determined by the Limulus Ameboyte Lysate assay. The VPBD and the VLC were determined by CBCT-analysis and compared to periapical radiographs. Data were typed on a spreadsheet and statistical analyzed. At the baseline samples, CFU counts and endotoxins levels ranged from 0 - 8.16 CFU/mL and 1.75 -149 EU/mL, respectively. Both contents were reduced after BP and after RCM. Positive correlations were found between the levels of endotoxins and VPBD as well as with previous episode of pain. Thereby, a positive correlation was found between the bacterial CFU counts with the endotoxins contents...

Evaluation of apical leakage in root canals filled with different sealers / Avaliação do desempenho de diferentes cimentos obturadores no selamento apical de canais radiculares

Objective: To evaluate the apical leakage exhibited by root canals filled with gutta-percha points and three different endodontic sealers. Material and Methods: Thirty-five human molars were used and had their palatal (maxillary molars) and distal roots (mandibular molars) sectioned, standardized and instrumented with Mtwo rotary system. The roots were filled through active lateral condensation technique and divided into three groups (n=10), according to the endodontic sealer employed: G1- AH Plus, G2- Fill Canal, G3- MTA Fillapex. All groups were filled by gutta-percha points and endodontic sealer. Gutta-percha points were immersed into sodium hypochlorite for 24 h to achieve disinfection. After the filling procedure, the roots were immersed into Indian ink for posterior diaphanization and obtainment of the images through stereomicroscopy. By analyzing the images in Adobe Illustrator CS5 software, the level of apical leakage was determined. The data obtained were submitted to Kruskal Wallis and Dunn tests, with level of significance set at 5%. Results: Statistically significant differences were found between G1 and G3. G2 did not show statistically significant differences. G1 exhibited the smallest apical leakage mean (12.85), followed by G2 (109.84) and G3 (101.15). Conclusions: Root canal obturation with gutta-percha points and AH plus sealer through lateral condensation technique provided lower apical leakage rates than the other endodontic sealers evaluated

Objetivo: Avaliar a infiltração apical apresentada em canais radiculares obturados com guta-percha e três diferentes cimentos obturadores. Material e Métodos: Foram utilizados 35 molares humanos, os quais tiveram suas raízes linguais (molares superiores) e distais (molares inferiores) seccionadas, padronizadas e instrumentadas com o sistema rotatório Mtwo. As raízes foram obturadas através da técnica de condensação lateral ativa e divididas em 3 grupos (n=10), de acordo com o cimento obturador utilizado: G1- AH Plus, G2- Fill Canal, G3- MTA Fillapex. Todos os grupos foram obturados com o conjunto guta-percha e cimento obturador, sendo que a guta-percha utilizada permaneceu 24 h imersa em hipoclorito de sódio, para sua desinfecção. Após a obturação, as raízes foram imersas em corante tinta da Índia, para posterior diafanização e obtenção de imagens através de estereomicroscópio. Através da análise das imagens no programa Adobe Illustrator CS5 foi determinado o nível de infiltração apical. Os dados obtidos foram submetidos aos testes estatísticos de Kruskal Wallis e Dunn, com nível de significância 5%. Resultados: Verificou-se presença de diferença estatística significante entre o G1 e G3, sendo que G2 não diferiu estatisticamente dos demais grupos. O G1 apresentou a menor média de infiltração apical (12.85), seguida pelo G2 (109.84) e G3 (101.15). Conclusões: A obturação de canais radiculares com cones de guta-percha e cimento AH plus através da técnica de condensação lateral proporciona baixos índices de infiltração apical, quando comparada aos demais cimentos obturadores avaliados

Detecção e ação antimicrobiana e antiendotóxica do extrato glicólico de gengibre utilizado como substância química auxiliar durante o retratamento endodôntico / Detection and antimicrobial action and antiendotoxic of glycolic ginger extract used as auxiliary chemical substance during endodontic retreatment

O presente estudo teve como objetivo detectar espécies bacterianas, quantificar microrganismos e endotoxinas em canais radiculares com insucesso endodôntico e presença de lesão periapical, correlacionando-os com os sinais e sintomas clínicos e, avaliar a ação do hipoclorito de sódio 1% e extrato glicólico de gengibre 20% sobre estes microrganismos e endotoxinas. Foram selecionados para o estudo dez dentes tratados endodonticamente com lesões periapicais persistentes que foram divididos em 2 grupos (n=5), de acordo com a substância química auxiliar utilizada durante o preparo biomecânico (PBM): hipoclorito de sódio 1% e extrato glicólico de gengibre 20% intercalado com solução salina fisiológica. Foram realizadas coletas do conteúdo do canal radicular imediatamente após a desobturação do dente, imediatamente após a instrumentação e, imediatamente após 14 dias da ação da medicação intracanal (MIC) de hidróxido de cálcio. Para todas as coletas foram realizados os seguintes testes: a)avaliação da atividade antimicrobiana por cultura microbiológica e através do método molecular - PCR; b) quantificação de endotoxinas verificada pelo lisado de amebócitos de Limulus. Os resultados foram submetidos a análise estatística de Wilcoxon e Mann-Whitney e mostraram que tanto o PBM quanto à MIC foram capazes de reduzir a quantidade de microrganismos e de endotoxinas, independente da solução utilizada. No entanto, endotoxinas não foram completamente neutralizadas. Espécies de E. faecalis, T. denticola, T. forsythia, P.endodontalis, P. gingivalis, P. nigrescens, P. intermedia, P. micra foram detectadas nos canais radiculares. Observou-se correlação positiva entre a quantidade de endotoxinas e o diâmetro das lesões. Concluiu se que as substâncias testadas foram eficazes na redução de microrganismos e de endotoxinas

The aim of the present study was to detect bacterial species, quantify microorganisms and endotoxins within failed root canals presenting periapical lesion, to correlate them with clinical signs and symptoms,and to evaluate the effect of 1% sodium hypochlorite and 20% glycolicginger extract on microorganisms and endotoxins. Ten endodontically treated teeth presenting persistent periapical lesion were selected for this study and divided into 2 groups (n=5), according to the auxiliary chemical substance employed during the biomechanical preparation(BMP): 1% sodium hypochlorite and 20% glycolic ginger extract interposed with saline solution. Root canal contents were collected right after root canal filling removal, right after instrumentation, and 14 day sof calcium hydroxide intracanal medication (ICM) activity. The following tests were performed for every collection: a) antimicrobial activity evaluation by means of microbiologic culture and molecular biology –PCR; b) endotoxins quantification assessed by Limulus amebocytelysate. The results were submitted to statistical analysis by Wilcoxon and Mann-Whitney tests. Both BMP and ICM were able to reduce microorganisms and endotoxins quantity, regardless the employed solution. However, endotoxins were not completely neutralized. E.faecalis, T. denticola, T. forsythia, P. endodontalis, P. gingivalis, P.nigrescens, P. intermedia, P. micra species were detected within root canals. A positive correlation was detected for the endotoxin quantityand the diameter of lesion. It can be concluded the tested substances were efficient for endotoxin and microorganisms reduction