Effect of different disinfection protocols in bacterial viability of an intraradicular biofilm formed in situ

Abstract The present study aimed to evaluate bacterial viability after the use of different disinfection protocols in root canals infected with a multispecies biofilm (MB) formed in situ. Palatal roots with a single canal were obtained from extracted maxillary molars and sterilized before being inserted into the mouth. The roots were contaminated with a MB in an intraoral appliance worn by ten volunteers. All volunteers wore six roots simultaneously in two intraoral devices for 21 days. One root from each volunteer was assigned to each group (n=10): PUI - passive ultrasonic irrigation; EC - Easy Clean; XPF - XP-endo Finisher; aPDT - antimicrobial photodynamic therapy; CI - conventional irrigation; and NC - negative control. The samples were evaluated under confocal laser scanning microscopy. The percentage of viable cells (VC) was calculated over the total percentage of MB biovolume. Data were statistically analyzed (α=5%). The cell viability in the entire root canal or for each third was compared between groups (Kruskal-Wallis test, Dunn post-hoc test) and for the same group (Friedman test, Dunn post-hoc test). Disinfection protocols were not significantly different from each other (P>.05). Samples in EC, PUI, and aPDT had lower cell viability than in NC (P<.05). In the coronal third of samples in the EC, XPF, PUI and aPDT, the percentage of VC biovolume was lower than in the NC (P<.05). The percentage of VC in EC samples was lower in the coronal and middle thirds than in the apical third (P<.05). EC, PUI and aPDT had significant effects on cell viability in intraradicular multispecies biofilm formed in situ when compared with untreated samples.

Resumo O presente estudo teve como objetivo avaliar a viabilidade bacteriana após o uso de diferentes protocolos de desinfecção em canais radiculares infectados com um biofilme multiespécies (MB) formado in situ. Raízes palatinas com canal único foram obtidas de molares superiores extraídos e esterilizadas antes de serem inseridas na boca. As raízes foram contaminadas com MB em um aparelho intraoral usado por dez voluntários. Todos os voluntários usaram seis raízes simultaneamente em dois dispositivos intrabucais por 21 dias. Uma raiz de cada voluntário foi atribuída a cada grupo (n=10): PUI - irrigação ultrassônica passiva; EC - Easy clean; XPF - XP-endo Finisher; aPDT - terapia fotodinâmica antimicrobiana; IC - irrigação convencional; e, NC - controle negativo. As amostras foram avaliadas em microscopia confocal de varredura a laser. A porcentagem de células viáveis (VC) foi calculada sobre a porcentagem total do biovolume de MB. Os dados foram analisados estatisticamente (α=5%). A viabilidade celular em todo o canal radicular ou em cada terço foi comparada entre os grupos (teste de Kruskal-Wallis, teste post-hoc de Dunn) e no mesmo grupo (teste de Friedman, teste post-hoc de Dunn). Os protocolos de desinfecção não foram significativamente diferentes entre si (P>0,05). Amostras dos grupos EC, PUI e aPDT apresentaram menor viabilidade celular do as do NC (P<0,05). No terço cervical das amostras do EC, XPF, PUI e aPDT, a porcentagem de biovolume de VC foi menor do que no NC (P<0,05). A porcentagem de VC nas amostras do EC foi menor nos terços cervical e médio do que no terço apical (P<0,05). EC, PUI e aPDT tiveram efeitos significativos na viabilidade celular do biofilme multiespécies intrarradicular formado in situ quando comparado com amostras não tratadas. Estudos clínicos devem investigar o papel da redução de cargas bacterianas viáveis no sistema de canais radiculares para o sucesso do tratamento endodôntico.

XP Endo Finisher-R and PUI as supplementary methods to remove root filling materials from curved canals

Abstract: This study assessed the ability of XP-endo Finisher R (FKG, La Chaux-de-Fonds, Switzerland) to remove filling remnants from curved mesiobuccal canals of maxillary molars, using the passive ultrasonic irrigation (PUI) technique as a comparison. Twenty-four curved main mesiobuccal canals (MB1) of maxillary molars were instrumented with Wave One (#25/07) and filled with gutta-percha points and AH Plus Sealer. Samples were then re-treated with a standardized protocol with Wave One (#35/06) as the master apical file. Micro-CT scans measured baseline volume of remaining filling material (in mm3). Samples were divided into two groups (n = 12) according to the supplementary cleaning approach: (PUI) or XP-endo Finisher R. Statistics compared baseline and final volume of filling material (within-group); and the percentage of filling material reduction (between-group). Mean baseline volumes, final volumes, and percentages of reduction (%) of filling material for XP-endo Finisher R and PUI were respectively: 0.060 mm3, 0.042 mm3, and 31.28%; and 0.064 mm3, 0.054 mm3, and 16.57%. Both tested protocols reduced the amount of filling material (p < 0.05). XP-endo Finisher R had higher percentage of reduction as compared to PUI (p < 0.05). XP-endo Finisher R and PUI used as supplementary cleaning protocols during re-treatment improved the removal of root filling material in curved canals; but XP-endo Finisher R was approximately twice more efficient. The complete filling material removal during re-treatment procedures is still a challenge. Supplementary cleaning protocols may help to remove the remaining material after the complete mechanical preparation of curved canals. XP-endo Finisher R was approximately twice more efficient than PUI.

Marginal gaps and internal voids after root-end filling using three calcium silicate-based materials: A Micro-CT analysis

Abstract This study evaluated the 3D quality of root-end filling, assessing the presence (volume and percentage) of marginal gaps and internal voids formed after retro-filling with three calcium silicate-based materials: MTA Angelus (Angelus Soluçoes Odontologicas, Londrina, PR, Brazil), Biodentine (Septodont Ltd., Saint Maur-des-Faussés, France) and Neo MTA Plus (Avalon Biomed Inc., Bradenton, Florida, US). Thirty human, extracted, single rooted teeth were used. Orthograde root canal treatment, root resection (3mm shorter than the apex) and retrograde cavity preparation with ultrasonic tips were performed. Teeth were divided into 3 groups (n =10 each) following a stratified randomization according to the initial volume of the root-end cavity. After retrofilling, samples were stored for 7 days. Then, two rounds of micro-CT scans were performed: soon after root-end preparation (with the cavity still empty) and 7 days after root-end filling. Marginal gaps, internal voids volume (mm3 and %), as well as, the overall defects (sum of gaps and voids) were evaluated. Statistics compared the three groups in relation to those defects. There was not statistical difference between groups regarding the marginal gaps (P≥ 0.05), the internal voids (P≥ 0.05), and the overall defects (P≥ 0.05). Median (mm3) and % of overall air-entrapment defects (gaps and/or voids) was: 0.004mm3 and 1.749% for MTA Angelus, 0.018mm3 and 6.660% for Biodentine, and 0.012mm3 and 4.079% for Neo MTA Plus. All materials had gaps and/or voids. No differences were found between MTA Angelus, Biodentine and Neo MTA Plus.

Resumo Este estudo avaliou a qualidade 3D de retro-obturações, avaliando a presença (volume e percentagem) de "gaps" marginais e "voids" internos formados a partir da retro-obturação com três materiais à base de silicato de cálcio: MTA Angelus (Angelus Soluções Odontológicas, Londrina, PR, Brasil), Biodentine (Septodont Ltd., Saint Maur-des-Faussés, França) and Neo MTA Plus (Avalon Biomed Inc., Bradenton, Flórida, US). Trinta dentes humanos unirradiculares extraídos foram usados. Após tratamento de canal ortrógrado, resecção radicular (3 mm apicais), o prepare da cavidade retrógrada com pontas ultrassônicas foi realizado. Os dentes foram divididos em três grupos (n= 10 cada) seguindo uma randomização estratificada, de acordo com o volume inicial da cavidade retrógrada. Depois da retro-obturação as amostras foram armazenadas por 7 dias. Duas rodadas de escaneamento de micro-CT foram realizadas: logo após o preparo retrógrado (com a cavidade ainda vazia) e 7 dias após a retro-obturação. O volume de "gaps" marginais e de "voids" internos (mm3 e %), bem como, os defeitos totais (soma de "gaps" e "voids") foram avaliados. Não houve diferença estatística entre os grupos em relação aos "gaps" marginais (P≥ 0,05), "voids" internos (P≥ 0,05), e aos defeitos totais (P≥ 0,05). A mediana (mm3) e porcentagem dos defeitos totais de aprisionamento de ar foi 0,004mm3 e 1,749% para o MTA Angelus, 0,018mm3 e 6,660% para o Biodentine, e 0,012mm3 e 4,079% para o Neo MTA Plus. Todos os materiais tiveram defeitos ("gaps"e/ou "voids"). Não houve diferença entre MTA Angelus, Biodentine e Neo MTA Plus.

Influence of photobiomodulation therapy on root development of rat molars with open apex and pulp necrosis

Abstract This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.