Shear Bond Strength and Film Thickness of a Naturally Antimicrobial Modified Dental Luting Cement.

Although several natural plants and mixtures have been known and used over the centuries for their antibacterial activity, few have been thoroughly explored in the field of dentistry. Thus, the aim of this study was to enhance the antimicrobial activity of a conventional glass ionomer cement (GIC) with natural plant extracts. The effect of this alteration on the bond strength and film thickness of glass ionomer cement was evaluated and related to an 0.5% chlorohexidine modified GIC. Olive leaves (Olea europaea), Fig tree (Ficus carica), and the leaves and roots of Miswak (Salvadora persica) were used to prepare an alcoholic extract mixture. The prepared extract mixture after the evaporation of the solvent was used to modify a freeze-dried glass ionomer cement at three different extracts: water mass ratios 1:2, 1:1, and 2:1. An 0.5% chlorhexidine diacetate powder was added to a conventional GIC for the preparation of a positive control group (CHX-GIC) for comparison. The bond strength to dentine was assessed using a material-testing machine at a cross head speed of 0.5 mm/min. Failure mode was analyzed using a stereomicroscope at 12× magnification. The cement film thickness was evaluated in accordance with ISO standard 9917-1. The minimum number of samples in each group was n = 10. Statistical analysis was performed using a Kruskal-Wallis test followed by Dunn's post hoc test for pairwise comparison. There was a statistically insignificant difference between the median shear bond strength (p = 0.046) of the control group (M = 3.4 MPa), and each of the CHX-GIC (M = 1.7 MPa), and the three plant modified groups of 1:2, 1:1, 2:1 (M = 5.1, 3.2, and 4.3 MPa, respectively). The CHX-GIC group showed statistically significant lower median values compared to the three plant-modified groups. Mixed and cohesive failure modes were predominant among all the tested groups. All the tested groups (p < 0.001) met the ISO standard of having less than 25 µm film thickness, with the 2:1 group (M = 24 µm) being statistically the highest among all the other groups. The plant extracts did not alter either the shear bond strength or the film thickness of the GIC and thus might represent a promising additive to GICs.

Potential for direct application of blue light for photo-disinfection of dentine.

The direct application of light for photo-disinfection potentially provides a safe and novel modality to inhibit or eliminate cariogenic bacteria residing upon and within dentine. This study aimed to both; characterize the pattern of transmission of 405 nm light through molar dentine at different tooth locations, as well as, determine the irradiation parameters that are antibacterial for Streptococcus mutans under various growth conditions, including lawns, planktonic cultures, and biofilms. To determine the amount of light (405 nm) transmitted at different anatomical tooth locations; irradiance values were recorded after blue light (470-4054 mW/cm2) had traversed through occlusal, oblique, and buccal dentine sections; and three thicknesses - 1, 2 and 3 mm were investigated. To determine tubular density; scanning electron micrographs from 2 mm outer (dentine-enamel junction) and inner (pulp) dentine sections were analysed. For photo-disinfection studies; S. mutans was irradiated using the same 405 nm wavelength light at a range of doses (110-1254 J/cm2) in both biofilm and planktonic cultures. The inhibitory effect of the irradiation on bacterial lawns was compared by measuring zones of inhibition; and for planktonic cultures both spectrophotometric and colony forming unit (CFU) assays were performed. A live/dead staining assay was utilised to determine the effect of irradiation on bacterial viability in mature biofilms. Data indicated that increasing dentine thickness decreased light transmission significantly irrespective of its orientation. Occlusal and oblique samples exhibited higher transmission compared with buccal dentine. Oblique dentine 405 nm light transmission was comparable with that of occlusal dentine independent of section thickness. An increased tubule density directly positively correlated with light transmission. Irradiation at 405 nm inhibited S. mutans growth in both biofilm and planktonic cultures and a dose response relationship was observed. Irradiation at doses of 340 and 831 J/cm2 led to significant reductions in bacterial growth and viability; as determined by CFU counting and live/dead staining. Data suggests that phototherapy approaches utilising a 405 nm wavelength have therapeutic potential to limit cariogenic bacterial infections both at the surface and within dentine.

Lactobacillus plantarum lipoteichoic acid disrupts mature Enterococcus faecalis biofilm.

Apical periodontitis is caused by biofilm-mediated root canal infection. Early phase oral bacterial biofilms are inhibited by Lactobacillus plantarum lipoteichoic acid (Lp.LTA). However, mature biofilms that develop over 3 weeks are more resistant to traditional endodontic medicaments. Therefore, this study examined the effectiveness of Lp.LTA on disrupting mature Enterococcus faecalis biofilms, and on enhancing the effects of endodontic medicaments. LTA was purified from L. plantarum through butanol extraction followed by hydrophobic and ion-exchange chromatography. E. faecalis biofilms were formed over 3 weeks on glass bottom dishes and in dentin blocks obtained from human single-rooted premolars. These mature biofilms were treated with or without Lp.LTA for 1 h, followed by additional treatment with either chlorhexidine digluconate (CHX), calcium hydroxide (CH), or triple antibiotics for 24 h. Biofilms on glass were live/dead stained and quantified by ZEN through confocal laser microscopy. Bio-films in dentin were fixed, sputter coated and analyzed by ImageJ with scanning electron microscopy. Preformed E. faecalis mature biofilms on the culture dishes were dose-dependently disrupted by Lp.LTA. Lp.LTA potentiated the effects of CHX or CH on the disruption of mature biofilm. Interestingly, CHX-induced disruption of preformed E. faecalis mature biofilms was synergistically enhanced only when pre-treated with Lp.LTA. Furthermore, in the dentin block model, Lp.LTA alone reduced E. faecalis mature biofilm and pre-treatment with Lp.LTA promoted the anti-biofilm activity of CHX. Lp.LTA could be an anti-biofilm or supplementary agent that can be effective for E. faecalis-biofilm-induced diseases.

Effects of Erbium:Yttrium-Aluminum-Garnet Laser Irradiation on Bovine Dentin Contaminated by Cariogenic Bacteria.

Objective: This study was performed to determine the bactericidal effects of erbium:yttrium-aluminum-garnet (Er:YAG) laser irradiation and the morphological and chemical composition changes in bovine dentin. Methods: Dentin slabs were prepared from bovine incisors, and then cultured with Streptococcus mutans to produce bacteria-infected dentin samples. The samples were randomly divided into five groups with Er:YAG laser irradiation energy densities of 0, 6.37, 12.73, 19.11, and 25.47 J/cm2. After irradiation, samples were stained and observed by confocal laser scanning microscopy. The bactericidal abilities were measured using live/dead staining. The morphology and chemical components were investigated by scanning electron microscopy and energy-dispersive spectrometry. Results: After irradiation, the elimination of bacteria and the smear layer were significantly better in the high energy density groups (19.11, 25.47 J/cm2) than in the low energy density groups (6.37, 12.73 J/cm2; p < 0.001). On morphological examination, the group with minimum energy density (6.37 J/cm2) showed superficial melting. In the high energy density groups (12.73, 19.11, and 25.47 J/cm2), laser-irradiated dentin showed a clean surface with open orifices. Significant increases were observed in the weight percentages of calcium (from 19.75 ± 0.69 to 34.47 ± 2.91, p < 0.001) and phosphate (from 8.58 ± 0.43 to 15.10 ± 1.81, p < 0.001), whereas significant decreases were observed for oxygen (from 49.84 ± 0.69 to 36.39 ± 2.86, p < 0.001) and carbon (from 26.06 ± 3.58 to 12.80 ± 2.26, p < 0.01) with increasing energy density. Conclusions: This study confirmed that Er:YAG laser irradiation has bactericidal and dentin conditioning effects.

Positive influence of simvastatin used as adjuvant agent for cavity lining.

OBJECTIVES: To assess the biological, antimicrobial, and mechanical effects of the treatment of deep dentin with simvastatin (SV) before application of a glass-ionomer cement (GIC). MATERIALS AND METHODS: Dentin discs were adapted to artificial pulp chambers and SV (2.5 or 1.0 mg/mL) was applied to the occlusal surface, either previously conditioned or not with EDTA (±EDTA). The extracts (culture medium + SV that diffused through dentin) was obtained and then applied to cultured odontoblast-like MDPC-23 cells. Cell viability, alkaline phosphatase (ALP) activity, and mineralization nodule (MN) deposition were evaluated. Untreated discs were used as control. The antibacterial activity of SV (2.5 or 1.0 mg/mL) against Streptococcus mutans and Lactobacillus acidophilus, as well as the bond strength of GIC to dentin in the presence of SV 2.5 mg/mL (±EDTA) were also assessed. The data were analyzed by ANOVA/Tukey tests (α = 5%). RESULTS: EDTA + SV 2.5 mg/mL significantly enhanced the ALP activity and MN deposition in comparison with the control, without changing in the cell viability (p < 0.05). The association EDTA + SV 2.5 mg/mL + GIC determined the highest ALP and MN values (p < 0.05). SV presented intense antimicrobial activity, and the EDTA dentin conditioning followed by SV application increased bond strength values compared with SV treatment alone (p < 0.05). CONCLUSION: SV presents antimicrobial activity and diffuses across conditioned dentin to biostimulate odontoblast-like pulp cells. CLINICAL SIGNIFICANCE: The use of SV as adjuvant agent for indirect pulp capping may biostimulate pulp cells thus preserving vitality and function of the pulp-dentin complex.

Curcumin-mediated antimicrobial photodynamic therapy reduces the viability and vitality of infected dentin caries microcosms.

BACKGROUND: To our knowledge, there is a lack of evidence on the effect of Antimicrobial Photodynamic Therapy (aPDT) by the application of curcumin against complex biofilms of dental caries lesions. This study aimed to evaluate the viability, vitality, and acid metabolism of infected dentin caries microcosms treated with curcumin-mediated aPDT. METHODS: After microcosm biofilms growing anaerobically on bovine dentin disks immersed in McBain medium with 1% sucrose at 37 °C for 5 days, the biofilms were treated by the association of DMSO water solution or 600 µmol L-1 curcumin with 0, 37.5 or 75 J cm-2 blue LED (455 nm). Then, the colony-forming units (CFU) counts of total microorganisms, total streptococci, mutans streptococci, and total lactobacilli were determined by plating. The lactic acid concentration was analyzed by enzymatic spectrophotometry method, while the vitality of intact biofilms was evaluated by confocal laser scanning microscope (CLSM). Statistical analysis was performed by Kruskal Wallis and post-hoc Dunn's tests (P < 0.05). RESULTS: Curcumin alone did not affect the viability of microorganisms and the vitality of intact biofilms. However, 75 J cm-2 LED alone decreased the total microorganisms and total lactobacilli counts. The combination of curcumin and LED reduced significantly the counts of all microorganism groups and the vitality of intact biofilms. Differences were not observed between the lactic acid concentrations of distinct groups. CONCLUSIONS: Therefore, curcumin-mediated aPDT was effective in reducing the viability and the vitality of infected dentin caries microcosms, without interfering in their acidogenicity.

Antimicrobial Potential of Laser Diode in Infected Dentin.

AIM: To evaluate the antibacterial effect of diode laser, associated or not with 2.5% sodium hypochlorite (NaOCl), against Enterococcus faecalis. MATERIALS AND METHODS: Eighty dentin blocks were obtained from single-rooted human teeth and sterilized. Seventy were inoculated with 0.01 mL of fresh bacterial inoculum (within 24 hours of preparation from pure culture) standardized to 1 McFarland turbidity. Contaminated blocks were incubated for 7 days at 37°C in humid conditions. Ten uncontaminated samples were incubated at 37°C during the contamination period to serve as a negative control group, while 10 of the infected specimens served as a positive control group. The dentin blocks were randomly divided into eight experimental groups (n = 10 each) according to the method of decontamination: 2.5% NaOCl alone; 2.5% NaOCl + photodynamic therapy (PDT) with methylene blue/660 nm laser at 18 J for 180 seconds; 2.5% NaOCl + PDT with methylene blue/660 nm laser at 8 J for 80 seconds; methylene blue alone; PDT alone with methylene blue/660 nm laser at 18 J for 180 seconds; PDT alone with methylene blue/660 nm laser at 8 J laser for 80 seconds; positive control group; and negative control group. Microbial growth was evaluated by culture medium turbidity and microbial concentration was analyzed by UV spectrophotometry (adjusted to read at wavelength l = 600 nM). RESULTS: Root canals treated with laser alone at 18 J for 180 seconds had higher bacterial contamination compared with groups in which NaOCl was used, with or without laser irradiation at 18 J for 180 seconds (p < 0.05). CONCLUSION: Photodynamic therapy with a 660 nm diode laser effectively reduced E. faecalis contamination. These findings can guide development of further studies in search of better alternatives for endodontic treatment. CLINICAL RELEVANCE: Chemical and mechanical root canal preparation plays an essential role in reducing microbial burden. However, microorganisms present in areas not mechanically reachable by endodontic instruments. As an alternative to fix this problem, the laser can be applied.

Bactericidal Effect of Various Laser Irradiation Systems on Enterococcus faecalis Biofilms in Dentinal Tubules: A Confocal Laser Scanning Microscopy Study.

OBJECTIVE: The aim of this study was to compare the bactericidal effect of various laser irradiation systems on Enterococcus faecalis biofilms in dentinal tubules by using a novel dentin infection model and confocal laser scanning microscopy (CLSM). BACKGROUND DATA: Laser-activated irrigations have been proposed as an adjuvant to conventional protocols of root canal treatment to enhance the smear layer removal, which is a promising protocol for root canal disinfection. MATERIALS AND METHODS: E. faecalis were centrifuged into the dentinal tubules, cultured for 3 weeks, and then received 1- and 3-min treatments as follows: (A) 5.25% sodium hypochlorite (NaOCl) irrigation, (B) Nd:YAG laser irradiation, (C) diode laser irradiation, (D) Nd:YAP laser irradiation, (E) Er,Cr:YSGG laser-activated NaOCl irrigation, and (F) Er:YAG laser-activated NaOCl irrigation. Bacterial reductions were assessed by CLSM using a LIVE/DEAD® bacterial viability stain method. RESULTS: For each group, the bacterial reduction increased as the treatment time increased (p < 0.05). The Er,Cr:YSGG and Er:YAG laser significantly enhanced the bactericidal effect of NaOCl (p < 0.05). Under the conditions of the same treatment time, bacterial reductions were presented in the descending order of Er:YAG + NaOCl, Er,Cr:YSGG + NaOCl > Nd:YAP > Nd:YAG, diode > NaOCl. CONCLUSIONS: Under the conditions of present study, treatments of Er:YAG + NaOCl and Er,Cr:YSGG + NaOCl presented the strongest bactericidal effect among the tested protocols and are potential protocols for root canal disinfection.

Antibacterial Effect of Surface Pretreatment Techniques against Streptococcus Mutans.

OBJECTIVE: The aim of this study was to evaluate the antibacterial surface pretreatment methods against Streptococcus mutans within the infected dentin surface using a tooth cavity model. MATERIAL AND METHODS: Seventy-two cavities were prepared on caries-free third molars (n = 8). After sterilization, teeth were inoculated with S. mutans for 48 h. One cavity of each tooth was used to evaluate the infection. Following inoculation, infected cavity surfaces were treated either with (1) Er:YAG Laser (1W; 5x5s, Smart 2940D Plus, Deka Laser), (2) Ozone (80s; HealOzone, Kavo), (3) ErYAG-Ozone combination, (4) Er:YAG-Ozone-CHX combination, (5) Chlorhexidine (CHX), (6) Clearfil Protect Bond (PB), (7) potassium-titanyl-phosphate (KTP) Laser (1W; 60 s, SMARTLITE D, Deka Laser), (8) KTP-Ozone combination, and (9) KTP-Ozone-CHX. Standardized amounts of dentin chips were obtained from the cavity walls, and the number of bacteria recovered was counted. Kruskal-Wallis test was used for statistical analyzes. RESULTS: Both sole antibacterial materials, CHX or Protect Bond application, exhibited the most effective antibacterial activity with 125 and 156 CFU is an acronym of "colony forming unit" usullay mentioned by acronym. (CFU/ml), respectively, among the groups evaluated (P < 0.05). Er:YAG laser irradiation and its combinations with other antibacterial surface pretreatment applications also inhibited the bacterial growth with, respectively, 1444, 406, and 294 CFU/ml bacterial recovery being more efficient than KTP laser irradiation and ozone combinations. CONCLUSIONS: As an alternative device with photodynamic effects, Er:YAG and KTP laser irradiations and their further combinations during the cavity pretreatment procedure with chlorhexidine and ozone treatments exerted antibacterial effect against S. mutans, whereas chlorhexidine and antibacterial dentin bonding application solely have the highest antibacterial effects.

Antimicrobial activity of Psidium cattleianum associated with calcium hydroxide against Enterococcus faecalis and Candida albicans: an in vitro study.

OBJECTIVE: Evaluate, in vitro, the antimicrobial activity of Psidium cattleianum leaf extracts combined with calcium hydroxide against Enterococcus faecalis and Candida albicans biofilm. MATERIALS AND METHODS: Dentin specimens obtained from extracted bovine incisors were infected during 14 days with E. faecalis ATCC 29212 and C. albicans ATCC 10231. The specimens were filled with calcium hydroxide pastes prepared with the following vehicles: Psidium cattleianum ethanolic, Psidium cattleianum propylene glycolic, distilled water, and saline as control. After 24 h, 3, 7, and 14 days, the canals were irrigated with sterile saline and dried. Dentin samples were collected from the canals with burs of increasing diameters. To determine the number of colony-forming units (CFU), samples were inoculated onto BHI agar supplemented with yeast extract (0.5%), at 37 °C, for 48 h, in CO2 enriched atmosphere. Comparisons among the groups for the variation factors were performed by ANOVA and Tukey's test. RESULTS: Ethanolic and propylene glycolic extracts showed significantly higher antimicrobial activity against E. faecalis (p < 0.01) when compared with distilled water. The ethanolic extract exhibited in 24 h the same antibacterial activity that propylene glycolic extract and distilled water after 7 and 14 days. For C. albicans, all were effective in reducing the number of CFU at all periods. CONCLUSION: The P. cattleianum ethanolic extract presented the fastest and highest antimicrobial activity against E. faecalis, significantly reducing the microbial load in 24 h. All medications were effective against C. albicans. CLINICAL RELEVANCE: The antibacterial potential of P. cattleianum and its biological compatibility associated with calcium hydroxide indicate promising applications in the field of dentistry.

Disinfection of dentinal tubules with 2% Chlorhexidine gel, Calcium hydroxide and herbal intracanal medicaments against Enterococcus faecalis: An in-vitro study.

AIM: This in vitro study was conducted to evaluate the disinfection of dentinal tubules using 2% Chlorhexidine gel, Honey, Aloe vera gel, Curcuma longa, Propolis gel and Calcium hydroxide against Enterococcus faecalis. MATERIALS AND METHOD: Two hundred and ten human mandibular first premolars were infected with Enterococcus faecalis for 21 days. Samples were divided into 7 groups. Group I- Saline (negative control), Group II- 2% Chlorhexidine gel(CHX), Group III- honey, Group IV- Aloe vera gel, Group V- 20% Curcuma longa gel, Group VI- Propolis gel and Group VII -Calcium hydroxide (CH). At the end of 1, 3 and 5 days, the antimicrobial efficacy of medicaments against E.faecalis was assessed at the depths of 200µm and 400µm. RESULTS: 2% Chlorhexidine gel was most effective followed by Propolis and Curcuma longa. CONCLUSION: 2% Chlorhexidine gel gave the best results. Among the herbal extracts Propolis and Curcuma longa hold a promising future but to implement their use as sole intracanal medicaments clinically, further in vivo and long term studies are warranted.

Ozone Gas Effect on Mineral Content of Dentin exposed to Streptococcus mutans Biofilm: An Energy-dispersive X-ray Evaluation.

AIM: This study aims to assess the effect of ozone gas on dentin exposed to Streptococcus mutans biofilm by evaluation of mineral content [log calcium-to-phosphorus (Ca/P)] using energy-dispersive X-ray (EDX) spectroscopy. MATERIALS AND METHODS: Five human third molars were sectioned into four slices of dentin and distributed in four groups: I - control (no treatment); II - ozone therapy; III - biofilm development; IV - ozone therapy followed by biofilm development. Mineral content (log Ca/P) was evaluated by EDX. Data were analyzed by analysis of variance and Tukey's test (p < 0.05). RESULTS: Results showed that the mineral content of control group (I) was similar to ozone group (II), and was statistically higher than biofilm (III) and ozone + biofilm (IV). The lowest log Ca/P was determined in biofilm group (III). CONCLUSION: It can be concluded that ozone gas did not grant preventive effects of demineralization by S. mutans biofilm on dentin surface. CLINICAL SIGNIFICANCE: Ozone gas therapy may be an alternative noninvasive treatment aiming to reduce the levels of caries-associated microorganisms. This therapy may, thereby, be an alternative and/or complementary treatment strategy in preventive dentistry.

Effect of the addition of chlorhexidine and miswak extract on the clinical performance and antibacterial properties of conventional glass ionomer: an in vivo study.

BACKGROUND: Antibacterial restorations can increase the success rate of minimum invasive dentistry especially in young permanent molars with deep carious lesions as an attempt to preserve maximum dental structure and avoid pulp exposure. Further research is warranted to evaluate different antibacterial agents. OBJECTIVES: This study aimed to evaluate the efficacy of adding chlorhexidine gluconate (CHX) or aqueous miswak (Salvadora persica) extract on the clinical performance and in vivo antibacterial activity of conventional anhydrous glass ionomer cement (GIC). DESIGN: The study was a randomized clinical trial. Sixty young permanent molars, with deep carious lesions in 6- to 9-year-old children were included. After randomization, atraumatic restorative treatment (ART) or stepwise excavation was performed followed by bacterial sampling from the center of the remaining carious dentine in the floor of the pulp. GIC powder was mixed with 0.5% chlorhexidine gluconate liquid in group I; with 100% aqueous miswak in group II; and with deionized water in group III (control). Clinical performance for all groups was assessed at 3, 6, and 9 months. After 9 months, restorations were removed and a second bacterial sample was collected for Streptococcus mutans (S. mutans) quantification and analysis by real-time polymerase chain reaction (PCR) test. RESULTS: Results showed no statistically significant difference in the success rate of the three groups at the 3-month interval. At 6 and 9 months, however, restoration success was 75% then 60% in group I, 100% then 90% in group II, and 95% then 85% in control group. Group II and the control group showed statistically significant higher survival rates than group I. All groups showed reduction in S. mutans counts in underlying dentine, but the percent reduction was significantly higher in group I. (P < 0.001). CONCLUSIONS: The addition of CHX and miswak to GIC showed superior antibacterial properties than conventional GIC, without seriously affecting the clinical performance of the restoration until the 6-month follow-up, but failure significantly increased in terms of marginal defects at 9 months with CHX (group 1).

Antibacterial Activity of Two Chemomechanical Caries Removal Gels on Carious Dentin of Primary Teeth: An in vitro Study.

INTRODUCTION: Chemomechanical caries removal (CMCR) is an effective method of caries removal especially for primary teeth as they cause less discomfort when compared with conventional caries removal. The most significant thing about caries removal is the elimination of cariogenic bacteria. This study compares the antibacterial activity of two CMCR gels. MATERIALS AND METHODS: A total of 40 primary molar teeth with carious dentin were split along the long axis in a laboratory. Total viable count (TVC) was taken for the teeth before splitting as a measure of colony-forming units per milliliter (CFU/mL). Each half was treated with either Carisolv or Carie-Care CMCR gels. Clean dentin samples were evaluated for Streptococcus mutans (SM) and Lactobacillus acidophilus (LB) after removal of carious tissue using the caries removal gels using serial dilutions and incubating on specific agar plates. RESULTS: The results showed significant reduction in mean TVC after use of both the CMCR gels. Both gels reduced the CFU/mL of SM and LB to a significant level (p < 0.05). However, there was no significant difference between the antibacterial activities of the two CMCR gels. CONCLUSION: The CMCR gels (Carisolv and Carie-Care) significantly reduced the residual TVC as well as SM and LB in carious primary dentin. Both CMCR gels had a similar antibacterial activity on the carious dentin of primary teeth. CLINICAL SIGNIFICANCE: The CMCR gels tested have a significant antibacterial activity and can be effectively used for elimination of caries-causing bacteria in primary teeth.

Dentin Topographic Features following Chemomechanical Caries Removal in Primary Teeth.

AIM: Study the topographic features of dentin after caries removal with a chemomechanical agent (Papacarie) compared with the conventional drilling method. STUDY DESIGN: The sample included 7 exfoliated and extracted primary teeth with carious dentin lesions, not reaching the pulp. Each tooth was sectioned longitudinally through the center of the carious lesions into two halves. The teeth were then divided into two groups according to the method of caries removal. Following caries removal, dentin topography and the cut section were examined using the scanning electron microscope. RESULTS: Papacarie produced an irregular, porous, rough and globular dentin appearance. The dentin surfaces were generally free of smear layer, visible bacteria and the dentinal tubules were opened. The dentin cut surfaces showed patent dentinal tubules with open orifices. The drilling method created a smooth and amorphous surface with a continuous smear layer occluding the dentinal tubules. Numerous bacteria were also observed. The cut dentin surfaces showed patent dentinal tubules with their orifices plugged with smear layer. CONCLUSIONS: Papacarie produced a rough and porous surface with partial or complete removal of the smear layer and opened dentinal tubules, while the drill produced a smooth surface with uniform smear layer occluding the dentinal tubules.

Ação antimicrobiana da terapia fotodinâmica sobre Enterococcus faecalis realizada pela combinação do aparelho fotopolimerizador a LED e curcumina / Antimicrobial activity of the combination of LED curing light and curcumin on Enterococcus faecalis in photodynamic therapy

Este trabalho avaliou a ação antimicrobiana da terapia fotodinâmica (TFD) realizada pela combinação do aparelho fotopolimerizador (LED 430-480nm) e curcumina 20µM, além de sua associação com Gel Carbopol 2%. Fotos padronizadas, na ausência e presença do gel Carbopol no interior do conduto, previamente preparado, foram analisadas no software OriginLab para verificar a manutenção da intensidade de luz ao longo do canal. Na análise antimicrobiana, uma linhagem de ATCC 4083 do Enterococcus faecalis foi utilizada para formação de biofilme sobre a superfície de discos de dentina (4mm de diâmetro) e para contaminação intratubular de segmentos radiculares (15mm), ambos obtidos de raízes bovinas. A ação sobre o biofilme foi avaliada de acordo com os grupos (n=20): [BST]: sem tratamento (controle); [BC]: Curcumina; [BL]: LED; [BCL]: Curcumina + LED e [BAL]: Azul de metileno 0.05% + LASER (660nm). A ação intratubular foi avaliada nos grupos (n=8): [IST]: sem tratamento (controle); [ICL]: Curcumina + LED; [ICGL]: Curcumina + Gel Carbopol + LED e [IAL]: Azul de metileno 0.05% + LASER (660nm). Discos e segmentos foram analisados em Microscópio Confocal de Varredura a LASER para detectar a porcentagem de bactérias viáveis. Os dados obtidos, que apresentaram distribuição normal, foram analisados estatisticamente pelo teste ANOVA, a 1 critério, e para distribuição não normal, Kruskall- Wallis. Os resultados mostraram que a presença do gel Carbopol no conduto não contribuiu na manutenção da intensidade da luz LED. Sobre o biofilme, os grupos BAL e BCL apresentaram maior redução microbiana, sem diferença entre eles, enquanto que, na ação intratubular, o IAL apresentou a melhor ação antimicrobiana, com diferença estatística com os demais. Concluiuse que a TFD realizada com o fotopolimerizador e curcumina é tão eficaz quanto a realizada com LASER e azul de metileno. Entretanto, é necessário investigar meios que permitam a sua utilização no interior do conduto radicular, já que o Gel Carbopol não manteve a intensidade da luz ao longo do trajeto.(AU)

This work aimed to evaluate the antimicrobial action of photodynamic therapy (PDT) by the combination of light curing device (LED 430-480nm) and Curcumin 20 µM, and its in association with Carbopol Gel 2%. Standard photos in the absence and presence of Carbopol gel inside of the root canal previously prepared, were analyzed in OriginLab software to check the light intensity maintenance along the canal. In microbial analysis, a strain of Enterococcus faecalis ATCC 4083 was used to make biofilm on the surface of dentin disks (4 mm in diameter), and intratubular contamination of root segments (15 mm), both obtained from bovine roots. The action on the biofilm was evaluated according to groups (n = 20): [BNT]: no treatment (control); [BC]: Curcumin; [BL]: LED; [BCL]: Curcumin + LED and [BML]: methylene blue 0.05% + LASER (660nm). Intratubular action was evaluated in groups (n = 8): [INT]: no treatment (control); [ICL]: Curcumin + LED; [ICGL]: Curcumin Carbopol Gel + LED + and [IML]: methylene blue 0.05% + LASER (660nm). Disks and segments were analyzed in LASER Scanning Confocal Microscope to detect the percentage of viable bacteria. The data, which showed normal distribution, were statistically analyzed by ANOVA 1 WAY test, and for no normal distribution, Kruskall-Wallis. The results showed that presence of Carbopol gel in the root canal did not increase the maintenance of intensity of LED light. About the biofilm, BML and BCL had higher microbial reduction, no difference between them, while in action intratubular, IML presented the higher antimicrobial action, with statistical difference with the other. It was concluded that the PDT realized with curing light and Curcumin is as effective as LASER and methylene blue. However, it is necessary to investigate ways that allow its use inside the root canal, because the Carbopol gel did not maintain the intensity of light until apex.(AU)

Antimicrobial Effects of Novel Triple Antibiotic Paste-Mimic Scaffolds on Actinomyces naeslundii Biofilm.

INTRODUCTION: Actinomyces naeslundii has been recovered from traumatized permanent teeth diagnosed with necrotic pulps. In this work, a triple antibiotic paste (TAP)-mimic scaffold is proposed as a drug-delivery strategy to eliminate A. naeslundii dentin biofilm. METHODS: Metronidazole, ciprofloxacin, and minocycline were added to a polydioxanone (PDS) polymer solution and spun into fibrous scaffolds. Fiber morphology, mechanical properties, and drug release were investigated by using scanning electron microscopy, microtensile testing, and high-performance liquid chromatography, respectively. Human dentin specimens (4 × 4 × 1 mm(3), n = 4/group) were inoculated with A. naeslundii (ATCC 43146) for 7 days for biofilm formation. The infected dentin specimens were exposed to TAP-mimic scaffolds, TAP solution (positive control), and pure PDS (drug-free scaffold). Dentin infected (7-day biofilm) specimens were used for comparison (negative control). Confocal laser scanning microscopy was done to determine bacterial viability. RESULTS: Scaffolds displayed a submicron mean fiber diameter (PDS = 689 ± 312 nm and TAP-mimic = 718 ± 125 nm). Overall, TAP-mimic scaffolds showed significantly (P ≤ .040) lower mechanical properties than PDS. Within the first 24 hours, a burst release for all drugs was seen. A sustained maintenance of metronidazole and ciprofloxacin was observed over 4 weeks, but not for minocycline. Confocal laser scanning microscopy demonstrated complete elimination of all viable bacteria exposed to the TAP solution. Meanwhile, TAP-mimic scaffolds led to a significant (P < .05) reduction in the percentage of viable bacteria compared with the negative control and PDS. CONCLUSIONS: Our findings suggest that TAP-mimic scaffolds hold significant potential in the eradication/elimination of bacterial biofilm, a critical step in regenerative endodontics.

Antibiofilm efficacy of photosensitizer-functionalized bioactive nanoparticles on multispecies biofilm.

INTRODUCTION: Newer disinfection strategies based on antibacterial nanoparticles and photodynamic therapy (PDT) aim to eliminate residual biofilm bacteria during root canal treatment. The aim of the current study was to test the newly developed rose bengal-functionalized chitosan nanoparticles (CSRBnps) for their interaction/uptake with monospecies bacteria/biofilm and assess their antibiofilm efficacy on a multispecies biofilm model in vitro. METHODS: The interaction of CSRBnps with bacterial cells was conducted using atomic force microscopy. Their membrane-damaging effect was determined by measuring the absorbance at 260 nm (OD260nm) using Enterococcus faecalis. The penetration of CSRBnps into E. faecalis biofilms was evaluated using confocal laser scanning microscopy (CLSM). Multispecies biofilms of Streptococcus oralis, Prevotella intermedia, and Actinomyces naeslundii were grown on dentin sections for 21 days to assess the antibiofilm efficacy. The biofilms were subjected to PDT (60 J/cm(2)) using CSRBnps and rose bengal. The treated/untreated biofilms were examined under scanning electron microscopy and CLSM. RESULTS: The CSRBnps synthesized were 60 ± 20 nm and showed absorption spectra similar to rose bengal. Atomic force microscopy showed adherence of CSRBnps to bacteria, roughening of cell surface, and cell disruption after PDT. CSRBnp treatment resulted in significantly increased bacterial membrane damage (P < .05). CSRBnps exhibited deeper penetration into the biofilm structure. Scanning electron microscopy and CLSM confirmed the complete disruption of multispecies biofilm with a reduction in viable bacteria and biofilm thickness (P < .05). CONCLUSIONS: These novel photosensitizer functionalized bioactive nanoparticles with increased affinity to bacterial cell membrane, higher penetration into biofilm structure, and enhanced ability to eliminate clinically relevant multispecies bacterial biofilm present a potential antibiofilm agent for root canal disinfection.

Antibacterial activity of various root canal sealers and root-end filling materials in dentin blocks infected ex vivo with Enterococcus faecalis.

OBJECTIVE: The aim was to investigate the antibacterial activity of the root-end filling materials MTA and IRM, different endodontic sealers and calcium hydroxide [Ca(OH)2] in experimentally infected dentinal tubules. MATERIALS AND METHODS: Ninety-four human root segments were prepared and the root canals were enlarged to ISO size 90. After smear removal, the specimens were infected with Enterococcus faecalis for 3 weeks. The roots were divided into eight groups and filled either with MTA, IRM, Ca(OH)2, gutta-percha and EndoRez (ER)/GuttaFlow (GF)/AH Plus (AH+) or with Resilon and Epiphany (EpRe). One group of specimens was left unfilled for control. Half of the specimens were treated for 1 day and the other half for 7 days in humid conditions at 37°C. Dentin samples from each canal were collected by enlarging the canals to ISO size 150; thus a dentinal depth of 300 µm was sampled. The number of cultivable bacteria was determined for each specimen. Statistical significance was set to 5%. RESULTS: After 1-day or 7-days of treatment, compared to control, all materials (except ER and GF at day 7) significantly reduced the number of bacteria. At day 1 and day 7, no significant difference was found between ER and GF and between Ca(OH)2, AH+, EpRe, IRM and MTA. However, a significant difference was found between these two groups of materials (except between GF and EpRe at day 7). Significantly more bacteria were cultured in the ER, GF, EpRe and IRM groups at day 7 compared to day 1. CONCLUSIONS: All materials exerted varying degrees of antibacterial activity which generally tended to decrease with time. The most stable antibacterial effect throughout the 7-day period was for Ca(OH)2, AH+ and MTA.

Bacterial leakage in root canals filled with AH Plus and dentine bonding agents.

OBJECTIVE: The aim of this study was to compare the efficacy of different dentine adhesives in delaying the coronal bacterial leakage of Enterococcus faecalis in filled root canals. Materials and methods. Ninety-five lower incisors of patients >65 years of age were instrumented using the ProTaper system and were irrigated with 1 mL of 2.5% sodium hypochlorite (NaOCl) alternated with 1 mL 17% EDTA between each file change. Final irrigation was performed with 5 mL of 17% EDTA and then flushed with 5 mL of distilled water. The teeth were randomly divided into five experimental groups (n = 15/group) and one of the following dentine adhesives was applied: (1) AdheSE; (2) Excite DSC; (3) Clearfil Protect Bond; (4) One Coat 7.0; or (5) Control group without adhesive. After filling the root canals, the samples were mounted on a double chamber device to evaluate the bacterial filtration of E. faecalis during a period of 240 days. The results underwent non-parametric Kaplan-Meier survival analysis and comparisons among groups were done using the Log-Rank test. RESULTS: At 240 days, E. faecalis was detected in samples of all groups in the lower chamber. The highest survival value was obtained by One Coat 7.0, giving statistically significant differences from the other groups, whereas Clearfil Protect Bond, AdheSE and Excite DSC showed similar behaviours, likewise similar to the Control group. CONCLUSIONS: One Coat 7.0 adhesive system provides the longest survival value to delay E. faecalis coronal leakage in filled root canals.