Antibacterial activity, cytocompatibility and effect of Bio-C Temp bioceramic intracanal medicament on osteoblast biology.

AIM: To analyse the antimicrobial and biological properties of a new bioceramic intracanal medicament (Bio-C Temp), and to compare it with two calcium hydroxide-based intracanal medicaments (Calen® and UltraCal® XS). METHODOLOGY: The direct contact and the crystal violet tests were performed to assess the antimicrobial activity of intracanal medicaments against Enterococcus faecalis. The cytocompatibility and the effect of the medication on the biology of the human osteoblast-like cell line (Saos-2) were evaluated with methylthiazole tetrazolium (MTT), neutral red, alkaline phosphatase activity and mineralization (alizarin red) assays. The data were analysed using one-way anova and Tukey's tests, two-way anova and Bonferroni's tests, or Kruskal-Wallis and Dunn's tests (α = 0.05). RESULTS: Bio-C Temp had significantly less antibacterial activity and biofilm biomass reduction than the other intracanal medicaments (P < 0.05). There was no difference in the viability of Saos-2 exposed to the various intracanal medicaments, except regarding the 1 : 2 dilution, when the Bio-C Temp group had significantly lower cell viability than the UltraCal® XS and Calen® groups (P < 0.05). Bio-C Temp induced significantly greater ALP activity than the other intracanal medicaments (P < 0.05) at day 1. Calen® induced significantly greater deposition of mineralized nodules than the other intracanal medicaments (P < 0.05), and no difference was observed between Bio-C Temp and UltraCal® XS (P > 0.05). CONCLUSIONS: Bio-C Temp had similar cytocompatibility at higher dilutions, and higher or similar induction of ALP activity and deposition of mineralized nodules in comparison with Calen® and UltraCal® XS. However, it had significantly less antibacterial and antibiofilm activity than Calen® and UltraCal® XS.

Effects of octenidine applied alone or mixed with sodium hypochlorite on eukaryotic cells.

AIM: To assess the effects of octenidine dihydrochloride (OCT) on eukaryotic cells and the cytotoxicity of OCT associated with sodium hypochlorite - NaOCl (NaOCl/OCT). METHODOLOGY: L929 fibroblasts and human osteoblast-like cells (Saos-2) were exposed to 0.1% OCT, 2% CHX, 2.5% NaOCl, 5.25% NaOCl and mixtures of 5.25% NaOCl and 0.1% OCT (NaOCl/OCT) at 90 : 10, 80 : 20 and 50 : 50 ratios. Cell viability was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red (NR) assays; type of cell death, by flow cytometry; cytoskeleton, by actin and α-tubulin fluorescence; and alkaline phosphatase (ALP) activity, by thymolphthalein release. The data were analysed by two-way ANOVA and Bonferroni tests (α = 0.05). RESULTS: MTT and NR assays revealed that 0.1% OCT had the lowest cytotoxicity (P < 0.05), followed by 2% CHX (P < 0.05). The 2.5% NaOCl, NaOCl/OCT 80 : 20 and NaOCl/OCT 50 : 50 solutions had intermediate cytotoxicity. NaOCl 5.25% and NaOCl/OCT 90 : 10 had the highest cytotoxicity (P < 0.05). The OCT group had a higher percentage of viable cells than the NaOCl and CHX groups (P < 0.05), and induced apoptosis at higher doses. The cytoskeleton alterations were observed at 0.12%, 0.6% and 2.02% for the NaOCl, CHX and OCT groups, respectively. The solutions did not induce ALP activity. CONCLUSION: Octenidine dihydrochloride was less cytotoxic, induced apoptosis at higher doses, caused few changes in the cytoskeleton and did not induce alkaline phosphatase activity. In addition, octenidine dihydrochloride reduced the cytotoxicity of 5.25% NaOCl when combined at 20 and 50%.

Effect of immersion in distilled water or phosphate-buffered saline on the solubility, volumetric change and presence of voids within new calcium silicate-based root canal sealers.

AIM: To assess the effect of immersion in distilled water or phosphate-buffered saline (PBS) on the solubility, volumetric change and presence of voids of calcium silicate-based root canal sealers (TotalFill BC, Sealer Plus BC and Bio-C), in comparison with the gold standard epoxy resin-based sealer (AH Plus). METHODOLOGY: All properties were evaluated after immersion in distilled water or PBS. Solubility was determined by the percentage of mass loss, whereas volumetric change and presence of voids were evaluated by micro-computed tomography, after 7 days of immersion. The volumetric change and percentage of voids between the baseline (after setting) and the experimental period were calculated. Statistical analysis was performed using one-way anova and Tukey's or Student's t-tests (α = 0.05). RESULTS: The calcium silicate-based sealers had significantly greater solubility and volumetric loss than AH Plus, after immersion in distilled water or PBS (P < 0.05). Bio-C had the greatest solubility (P < 0.05), followed by TotalFill BC and Sealer Plus BC, which were similar (P > 0.05). Regarding the volumetric change, AH Plus had a volume increase, with similar values in distilled water and PBS (P > 0.05). TotalFill BC, Sealer Plus BC and Bio-C had a similar volumetric change (P > 0.05). The calcium silicate-based materials had the greatest solubility and volume loss after immersion in distilled water (P < 0.05). There was no difference in the percentage of voids amongst the sealers, before and after immersion in distilled water or PBS (P > 0.05). CONCLUSIONS: TotalFill BC, Sealer Plus BC and Bio-C had significantly greater solubility and volumetric loss than AH Plus. Although storage in PBS significantly reduced the solubility and volumetric change of calcium silicate-based sealers, their solubility remained above that recommend by ISO 6876. All the sealers evaluated had low and similar voids, even after immersion in distilled water or PBS.

Cytocompatibility, bioactive potential and antimicrobial activity of an experimental calcium silicate-based endodontic sealer.

AIM: To evaluate the cytocompatibility, bioactive potential, antimicrobial and antibiofilm activities of an experimental calcium silicate-based endodontic sealer, in comparison with TotalFill BC Sealer and AH Plus. METHODOLOGY: Cytocompatibility was assessed by methyltetrazolium (MTT) and neutral red (NR) assays, after exposure of the Saos-2 cells to the sealer extracts (1 : 2, 1 : 4, 1 : 8, 1 : 16 and 1 : 32 dilutions) for 24 h. The sealers were manipulated and placed in 12-well culture plates and exposed to ultraviolet light; then, 5 mL of DMEM without serum was added. Cell bioactivity was evaluated by alkaline phosphatase activity (ALP) and Alizarin red staining (ARS). Antimicrobial and antibiofilm activities were evaluated by direct contact test (DCT) on planktonic cells (DCTPC) and modified DCT on biofilm formed in bovine dentine blocks (MDCT). MTT, NR and ALP data were analysed by two-way anova and Bonferroni tests; ARS data by anova and Tukey's tests; and the microbiological data by Kruskal-Wallis and Dunn tests (α = 0.05). RESULTS: The experimental sealer, TotalFill BC and AH Plus were not cytotoxic to Saos-2, in comparison with the negative control (P > 0.05). Greater ALP was observed after 7 days of exposure of Saos-2 to AH Plus and the experimental sealer (P < 0.05) when compared to the control. Significantly greater mineralized nodule production was observed for TotalFill BC and the experimental sealer (P < 0.05). In DCTPC, the experimental sealer and TotalFill BC were associated with a significantly greater reduction of E. faecalis (P < 0.05) and eliminated C. albicans. In MDCT, the experimental sealer and TotalFill BC had significantly greater antibiofilm efficacy (P < 0.05). CONCLUSIONS: The experimental calcium silicate-based sealer was cytocompatible, bioactive, antimicrobial against E. faecalis and C. albicans and effective against E. faecalis biofilms, with potential for use in root canal treatment.

Effectiveness of several solutions to prevent the formation of precipitate due to the interaction between sodium hypochlorite and chlorhexidine and its effect on bond strength of an epoxy-based sealer.

AIM: To evaluate the effectiveness of isopropyl alcohol, saline or distilled water to prevent the precipitate formed between sodium hypochlorite (NaOCl) and chlorhexidine (CHX) and its effect on the bond strength of an epoxy-based sealer in radicular dentine. METHODOLOGY: The root canals of 50 extracted human canines (n = 10) were instrumented. In G1, root canals were irrigated with 17% EDTA and 2.5% NaOCl; G2, as G1, except that 2% CHX was used as the final irrigant. In the other groups, intermediate flushes with isopropyl alcohol (G3), saline (G4) or distilled water (G5) were used between NaOCl and CHX. The specimens were submitted to SEM analysis to evaluate the presence of debris and smear layer, in the apical and cervical segments. In sequence, fifty extracted human canines were distributed into five groups (n = 10), similar to the SEM study. After root filling, the roots were sectioned transversally to obtain dentine slices, in the cervical, middle and apical thirds. The root filling was submitted to a push-out bond strength test using an electromechanical testing machine. Statistical analysis was performed using Kruskal-Wallis and Dunn's tests (α = 5%). RESULTS: All groups had similar amounts of residue precipitated on the canal walls (P > 0.05). The push-out bond strength values were similar for all groups, independently of the root third evaluated (P > 0.05). CONCLUSIONS: Isopropyl alcohol, saline and distilled water failed to prevent the precipitation of residues on canal walls following the use of NaOCl and CHX. The residues did not interfere with the push-out bond strength of the root filling.

Antibacterial effectiveness of several irrigating solutions and the Endox Plus system - an ex vivo study.

AIM: To compare the ex vivo antibacterial effectiveness of the Endox Plus system and sodium hypochlorite (NaOCl) in combination with BioPure MTAD (Tulsa Dental, Tulsa, OK, USA) or with EDTA in Enterococcus faecalis-contaminated root canals. METHODOLOGY: After initial preparation, the root canals of 70 single-rooted human teeth were inoculated with E. faecalis (ATCC 29212) and incubated for 21 days. Specimens were divided into five groups: Endox Plus/saline; 2.5% NaOCl/MTAD; 2.5% NaOCl/EDTA; saline (positive control); negative control (root canals not prepared, nor irrigated). Samples were collected using paper points. Microbiological analysis evaluated the number of CFUs. Data were analysed by anova and Tukey tests at 0.05 significance. RESULTS: All specimens had bacterial growth after the incubation period, with similar CFU per mL counts (P > 0.05). After chemo-mechanical preparation, the number of bacteria in all groups reduced, except for the negative control. No significant differences were observed between 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA, but these groups had lower CFU counts than the other groups (P < 0.05). In the final samples, an increase in the bacterial counts was observed for Endox Plus/saline, 2.5% NaOCl/MTAD, 2.5% NaOCl/EDTA and saline (P < 0.05) with no significant differences between these groups. CONCLUSIONS: This ex vivo study revealed that the Endox Plus system was associated with a reduced antibacterial effectiveness compared with conventional irrigation using 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA. All irrigation procedures allowed recovery of bacteria 7 days after treatment, demonstrating persistence of contamination within the root canal system.