Effect of hydrogel-based antibiotic intracanal medicaments on crown discoloration.

OBJECTIVES: This study evaluated the effects of low and moderate concentrations of triple antibiotic paste (TAP) and double antibiotic paste (DAP) loaded into a hydrogel system on crown discoloration and explored whether application of an adhesive bonding agent prevented crown discoloration. MATERIALS AND METHODS: Intact human molars (n = 160) were horizontally sectioned 1 mm apical to the cementoenamel junction. The crowns were randomized into 8 experimental groups (calcium hydroxide, Ca[OH]2; 1, 10, and 1,000 mg/mL TAP and DAP; and no medicament. The pulp chambers in half of the samples were coated with an adhesive bonding agent before receiving the intracanal medicament. Color changes (ΔE) were detected by spectrophotometry after 1 day, 1 week, and 4 weeks, and after 5,000 thermal cycles, with ΔE = 3.7 as a perceptible threshold. The 1-sample t-test was used to determine the significance of color changes relative to 3.7. Analysis of variance was used to evaluate the effects of treatment, adhesive, and time on color change, and the level of significance was p < 0.05. RESULTS: Ca(OH)2 and 1 and 10 mg/mL DAP did not cause clinically perceivable tooth discoloration. Adhesive agent use significantly decreased tooth discoloration in the 1,000 mg/mL TAP group up to 4 weeks. However, adhesive use did not significantly improve coronal discoloration after thermocycling when 1,000 mg/mL TAP was used. CONCLUSIONS: Ca(OH)2 and 1 and 10 mg/mL DAP showed no clinical discoloration. Using an adhesive significantly improved coronal discoloration up to 4 weeks with 1,000 mg/mL TAP.

Effect of Hydrogel-Based Antibiotic Intracanal Medicaments on Push-Out Bond Strength.

OBJECTIVE: This study aimed to evaluate the effects of typical clinical concentration (1,000 mg/mL), low concentration (1 mg/mL) triple antibiotic pastes (TAP), and double antibiotic pastes (DAP) on the bond strength between various root cements and radicular dentin. MATERIALS AND METHODS: Intact single-rooted human teeth (n = 144) were horizontally decoronated and canals instrumented. The roots were treated for 4 weeks with Ca(OH)2, 1,000 mg/mL of TAP or DAP, and 1 mg/mL of TAP or DAP. Untreated roots served as a control. After treatment, the medicaments were irrigated and each group was divided into three subgroups receiving MTA, Biodentine, or Endosequence putty cement. After 2 weeks, coronal and middle root cylinders were obtained from each root. Push-out bond strength test and failure analysis were performed for all root cylinders. STATISTICAL ANALYSIS: Three-way ANOVA, pairwise comparisons and logistic regression were used for statistical analyses. A significance level of 5% was used. RESULTS: For MTA applied in the coronal part of the roots, 1 mg/mL DAP and TAP and Ca(OH)2 demonstrated significantly higher bond strength compared with the typical clinical concentration and the control groups. For Biodentine applied coronally in the roots, 1 mg/mL of DAP resulted in significantly higher bond strength than all other groups. For Endosequence putty cement applied coronally in the roots, 1 mg/mL of DAP offered significantly higher bond strength than all groups except for Ca(OH)2. CONCLUSION: The use of 1 mg/mL DAP resulted in significantly higher push-out bond strength compared with the typical clinical concentration of TAP and DAP regardless of the type of the root cement used.

Effects of radiopaque double antibiotic pastes on the proliferation, alkaline phosphatase activity and mineral deposition of dental pulp stem cells.

OBJECTIVE: The aim of this study was to investigate the effects of two radiopaque agents, barium sulfate (BaSO4) or zirconium oxide (ZrO2) in double antibiotic paste (DAP), on the proliferation and mineral deposition of human dental pulp stem cells (DPSC). MATERIALS AND METHODS: Radiopaque antimicrobial medicaments composed of methylcellulose (MC) thickening polymer with BaSO4 or ZrO2 and either 1 or 5 mg/mL DAP (equal portions of metronidazole and ciprofloxacin) were used to investigate DPSC proliferation after 3 days, and alkaline phosphatase (ALP) activity and mineral deposition after 7 and 14 days. Radiopaque agents without DAP and Ca(OH)2 were used as controls. RESULTS: MC-BaSO4 DAP and MC-ZrO2 DAP at 1 or 5 mg/mL had no adverse effect on DPSC proliferation, compared to the media and MC controls. MC-ZrO2 (DAP-free) greatly increased ALP activity after 7 days. DPSC mineral deposition was modestly reduced at 7 days by MC-BaSO4 DAP and MC-ZrO2 DAP, but not by DAP-free radiopaque agents, and was most reduced by 5 mg/mL DAP in the 14-day cultures. CONCLUSIONS: MC-BaSO4 or MC-ZrO2 medicaments containing up to 5 mg/mL of DAP supported the proliferation and early osteogenic differentiation of DPSC. Low DAP concentrations and short culture times led to more favorable effects on ALP activity and mineral deposition by DPSC. The findings suggest that radiopaque agents added for the purpose of detecting whether medicaments occupy the full extent of the root canal may have clinical applications. Radiopaque antibiotic medicaments containing low DAP concentrations may be an alternative to Ca(OH)2 for regenerative endodontic procedures.

Regenerative Endodontics: A Systematic Analysis of the Failed Cases.

INTRODUCTION: The aim of this systematic review was to analyze failed cases of regenerative endodontic treatment (RET) reported in the literature in terms of etiology, diagnosis, treatment protocols, signs of failure, and additional endodontic interventions. METHODS: Electronic searches were performed in PubMed, Web of Science, and ProQuest Dissertations & Theses databases. All in vivo publications in humans that reported at least 1 failed case of RET were included in this systematic review. Failed RET cases were defined in the current study as any case that required an additional endodontic intervention or extraction after the completion of the initial RET. RESULTS: A total of 28 studies that reported 67 failed cases of RET were included in this review. A total of 37 failed RET cases reported the etiology that resulted in the initiation of RET; 59% of these cases were caused by dental trauma, and 30% were caused by dens evaginatus. A total of 26 (39%) failed RET cases were detected at least 2 years after the initiation of RET. A total of 53 (79%) failed RET cases were presented with signs and/or symptoms of persistent infection. CONCLUSIONS: Persistent infection was the main presentation in 79% of failed RET cases. Furthermore, 39% of failed RET cases were identified after more than 2 years of follow-up. Future studies should include a detailed description of the etiology, preoperative variables, intraoperative protocols, and postoperative follow-up to provide a better understanding of failed cases after RET.

The antimicrobial properties, cytotoxicity, and differentiation potential of double antibiotic intracanal medicaments loaded into hydrogel system.

OBJECTIVE: This study evaluated the antimicrobial properties, cytotoxicity, and mineralization potential of methylcellulose hydrogels loaded with low concentrations of double antibiotic pastes (DAP). MATERIALS AND METHODS: The direct and residual antibacterial effects of 1, 5, and 10 mg/mL of DAP loaded into hydrogels as well as calcium hydroxide (Ca(OH)2) were tested against single-species biofilms of Enterococcus faecalis and dual-species biofilms (Enterococcus faecalis and Prevotella intermedia). The effects of DAP hydrogels on proliferation and mineralization of dental pulp stem cells (DPSC) were tested using MTT assays, alkaline phosphate activity (ALP), and alizarin red staining. Fisher's exact tests, Wilcoxon rank sum tests, and one-way ANOVA were used for statistical analyses (α = 0.05). RESULTS: All tested concentrations of DAP hydrogels as well as Ca(OH)2 demonstrated significant direct antibacterial effects against single- and dual-species biofilms. However, only 5 and 10 mg/mL of DAP hydrogels exhibited significant residual antibacterial effects against both types of tested biofilms. Only 1 mg/mL of DAP hydrogels did not have significant negative effects on DPSC viability, ALP activity, and mineralization nodule formation. However, 5 and 10 mg/mL of DAP hydrogels caused significant negative effects on cytotoxicity and mineralization nodule formation of DPSC. CONCLUSIONS: Hydrogels containing 1 mg/mL DAP offered significant direct antibacterial effects against single- and dual-species biofilms without causing significant negative effects on viability, ALP activity, and mineralization nodule formation of DPSC. CLINICAL RELEVANCE: The methylcellulose-based hydrogel proposed in this study can be used clinically as a biocompatible system to deliver controlled low concentrations of DAP.

The Radiopacity and Antimicrobial Properties of Different Radiopaque Double Antibiotic Pastes Used in Regenerative Endodontics.

INTRODUCTION: We evaluated the radiopacity and antibacterial properties of various concentrations of double antibiotic paste (DAP) containing barium sulfate (BaSO4) or zirconium oxide (ZrO2) radiopaque agents. METHODS: The radiopacity of 1, 10, and 25 mg/mL DAP containing 30% (w/v) BaSO4 or ZrO2, DAP-free radiopaque pastes, and commercially available radiopaque calcium hydroxide (Ca[OH]2) were evaluated according to ISO 6876/2001 with slight modifications (n = 6 per group). Dentin samples (n = 70) infected anaerobically for 3 weeks with bacterial biofilms obtained from a root canal of an immature tooth with pulpal necrosis were treated with similar experimental pastes or received no treatment (n = 7). After 1 week, the pastes were rinsed off, and biofilm disruption assays were conducted. To show the residual antibacterial effects, sterile dentin samples (n = 70) were pretreated for 1 week with the same pastes (n = 7). The pastes were rinsed off, and the samples were immersed in phosphate-buffered saline for 24 hours and infected anaerobically with the same bacterial biofilm mentioned earlier for 3 weeks before conducting biofilm disruption assays. Sterile dentin blocks were used in both antibacterial analyses as negative control groups (n = 7). Wilcoxon rank sum tests were used for statistical analyses. RESULTS: No tested concentrations of BaSO4 DAP or ZrO2 DAP showed significant differences from Ca(OH)2 in radiopacity. However, all tested concentrations of BaSO4 DAP, ZrO2 DAP, and Ca(OH)2 exhibited significant direct antibacterial effects. ZrO2 DAP at 1 mg/mL and Ca(OH)2 did not show significant residual antibacterial effects. CONCLUSIONS: BaSO4 DAP at 1 mg/mL provided significantly superior residual antibacterial effects and comparable radiopacity with the commercially available Ca(OH)2.

Antibacterial Effects of Antimicrobials Used in Regenerative Endodontics against Biofilm Bacteria Obtained from Mature and Immature Teeth with Necrotic Pulps.

INTRODUCTION: We investigated the direct and residual antibacterial effects of intracanal antimicrobials against bacterial biofilms obtained from infected mature and immature teeth with necrotic pulps. METHODS: Sterile dentin slabs (n = 100) were inoculated with bacterial biofilms obtained from root canals of an immature or a mature tooth with pulpal necrosis and incubated anaerobically for 3 weeks (n = 50 per biofilm). Dentin infected with each type of biofilm received 1 week of treatment with 1 or 5 mg/mL double antibiotic paste (DAP) in methylcellulose hydrogels, calcium hydroxide, or placebo paste or received no treatment (n = 10). The pastes were removed, and biofilm disruption assays were performed. Additional dentin slabs (n = 100) were pretreated with the same treatments (n = 20). The pastes were rinsed off, and the samples were immersed in phosphate-buffered saline for 1 week. Thereafter, samples from the treatment groups were infected with bacterial biofilm from both clinical sources mentioned earlier (n = 10 per biofilm) and incubated anaerobically for 3 weeks before conducting biofilm disruption assays. Uninfected dentin slabs were used for both antibacterial experiments as negative control groups (n = 20). RESULTS: All antimicrobials showed significant direct antibacterial effects regardless of the biofilm source. Dentin pretreated with 5 mg/mL DAP provided significantly higher residual antibacterial effects in comparison with all other groups regardless of the source of biofilm. Dentin pretreated with calcium hydroxide did not show any residual antibacterial effects. CONCLUSIONS: Tested antimicrobials showed significant direct antibacterial effects. Only 5 mg/mL DAP exhibited significant residual antibacterial effects against bacterial biofilms from an infected root canal of an immature tooth.

Direct and indirect antibacterial effects of various concentrations of triple antibiotic pastes loaded in a methylcellulose system.

We investigated the direct and indirect (residual) antibacterial effects of various concentrations of triple antibiotic paste (TAP) loaded into a methylcellulose system. Enterococcus faecalis (E. faecalis) was grown on sterilized dentin blocks (n = 60) and treated with clinically used TAP (1,000 mg/mL), low concentrations of methylcellulose-based TAP (100, 10, and 1 mg/mL), placebo paste, or 1.5% NaOCl (n = 10). The pastes were then removed, and biofilm disruption assays were performed. Additional dentin blocks (n = 120) were pretreated with the same experimental groups (n = 20). The pastes were rinsed off, and the samples were immersed independently in phosphate-buffered saline for 2 and 4 weeks (n = 10). E.faecalis was then grown on the dentin blocks, and biofilm disruption assays were performed. Fisher's Exact and Wilcoxon rank sum tests were used for statistical analyses. With regard to direct antibacterial effects, all treatment groups demonstrated complete eradication of biofilms in comparison to placebo paste, while 10 mg/mL of TAP or higher provided substantial residual antibacterial effects. However, dentin treated with 1 mg/mL of TAP or 1.5% NaOCl did not provide substantial residual antibacterial effects. Dentin pretreated with 10 mg/mL of TAP or higher exhibited extended residual antibacterial effects and can thus be used during endodontic regeneration.(J Oral Sci 58, 575-582, 2016).

Residual antibiofilm effects of various concentrations of double antibiotic paste used during regenerative endodontics after different application times.

OBJECTIVE: We investigated the residual antibiofilm effects of different concentrations of double antibiotic paste (DAP) applied on radicular dentin for 1 or 4 weeks. DESIGN: Dentin samples were prepared (n=120), sterilized and pretreated for 1 or 4 weeks with the clinically used concentration of DAP (500mg/mL), low concentrations of DAP (1, 5 or 50mg/mL) loaded into a methylcellulose system, calcium hydroxide (Ca(OH)2), or placebo paste. After the assigned treatment time, treatment pastes were rinsed off and the samples were kept independently in phosphate buffered saline for 3 weeks. Pretreated dentin samples were then inoculated with Enterococcus faecalis and bacterial biofilms were allowed to grow for an additional 3 weeks. Biofilms were then retrieved from dentin using biofilm disruption assays, diluted, spiral plated, and quantified. Fisher's Exact and Wilcoxon rank sum tests were used for statistical comparisons (α=0.05). RESULTS: Dentin pretreatment for 4 weeks with 5, 50 or 500mg/mL of DAP demonstrated significantly higher residual antibiofilm effects and complete eradication of E. faecalis biofilms in comparison to a 1 week pretreatment with similar concentrations. However, dentin pretreated with 1mg/mL of DAP or Ca(OH)2 did not provide a substantial residual antibiofilm effect regardless of the application time. CONCLUSIONS: Dentin pretreatment with 5mg/mL of DAP or higher for 4 weeks induced significantly higher residual antibiofilm effects in comparison to a 1 week pretreatment with the same concentrations.

Evaluation of selected properties of a new root repair cement containing surface pre-reacted glass ionomer fillers.

OBJECTIVE: This study evaluated selected properties of a prototype root repair cement containing surface pre-reacted glass ionomer fillers (S-PRG) in comparison to mineral trioxide aggregate (MTA) and intermediate restorative material (IRM). MATERIALS AND METHODS: The antibacterial effect of S-PRG, MTA, and IRM cements was tested against Porphyromonas gingivalis and Enterococcus faecalis after 1 and 3 days of aging of the cements. The set cements were immersed in distilled water for 4 h to 28 days, and ion-releasing ability was evaluated. Initial and final setting times of all cements were evaluated using Gilmore needles. The push-out bond strength between radicular dentin and all cements was tested at different levels of the roots. RESULTS: S-PRG and IRM cements, but not MTA cement, demonstrated significant antibacterial effect against P. gingivalis. All types of cements exhibited significant antibacterial effect against E. faecalis without being able to eliminate the bacterium. S-PRG cement provided continuous release of fluoride, strontium, boron, sodium, aluminum, and zinc throughout all tested time points. Both initial and final setting times were significantly shorter for S-PRG and IRM cements in comparison to MTA. The push-out bond strength was significantly lower for S-PRG cement in comparison to MTA and IRM at coronal and middle levels of the roots. CONCLUSIONS: S-PRG cement demonstrated significant antibacterial effects against endodontic pathogens, multiple ion-releasing ability, relatively short setting time, and low bonding strength. CLINICAL RELEVANCE: S-PRG cement can be used as a one-visit root repair material with promising antibacterial properties and ion-releasing capacity.

Effect of Antimicrobials Used in Regenerative Endodontic Procedures on 3-week-old Enterococcus faecalis Biofilm.

INTRODUCTION: We evaluated the effect of various antimicrobials used in endodontic regeneration on a 3-week-old Enterococcus faecalis biofilm. METHODS: E. faecalis biofilm was grown on standardized dentin samples for 3 weeks. Infected dentin samples were randomized into 8 experimental groups (n = 8) and treated with calcium hydroxide (Ca[OH]2), 500 mg/mL of double antibiotic paste (DAP, equal portions of metronidazole and ciprofloxacin), low dilutions of DAP (1 or 0.1 mg/mL loaded into a methylcellulose vehicle system), sterile saline, or placebo paste (only methylcellulose) for 7 days. The other experimental groups were treated with 1.5% sodium hypochlorite (NaOCl) or 2% chlorhexidine gluconate (CHX) solutions for 5 minutes. After the assigned treatments, the bacterial biofilms were detached from dentin, spiral plated, and quantified using an automated counting machine. Permutation tests followed by Sidak post hoc multiple comparisons were used for statistical analyses (α = 0.05). RESULTS: The infected dentin treated with 1.5% NaOCl or 500 mg/mL of DAP provided complete eradication of bacterial biofilm. Furthermore, the infected dentin treated with 2% CHX, Ca(OH)2, or 1 mg/mL of DAP had a comparable antibiofilm effect, but they were not able to completely eradicate bacterial biofilm. No significant difference in the antibiofilm effect was observed between 500 mg/mL of DAP, Ca(OH)2, 1.5% NaOCl, and 2% CHX. CONCLUSIONS: At least 1 mg/mL of DAP in a methylcellulose vehicle system is required to eliminate a substantial amount of E. faecalis biofilm. Furthermore, the antibiofilm effects of 1.5% NaOCl and 2% CHX irrigation solutions were comparable with that of 500 mg/mL of DAP and Ca(OH)2.

The effects of radicular dentine treated with double antibiotic paste and ethylenediaminetetraacetic acid on the attachment and proliferation of dental pulp stem cells.

AIM: This study explored the effects of dentine treated with two concentrations of double antibiotic paste (DAP) and ethylenediaminetetraacetic acid (EDTA) on the attachment and proliferation of dental pulp stem cells (DPSCs). MATERIALS AND METHODS: Radicular dentine samples were prepared with identical dimensions and randomized into six groups (n = 4). Four groups were treated with double antibiotic paste (DAP) at concentrations of 500 mg ml(-1) or 1 mg ml(-1) with or without EDTA. The other two groups were treated with EDTA only or received no treatment. DPSCs were seeded on each dentine sample (10 000 cells per sample). Lactate dehydrogenase activity assays were used to calculate the attached DPSCs after 1 day of incubation. Water soluble tetrazolium assays were performed to investigate DPSCs proliferation on the treated dentine samples after three additional days of incubation. Two-way anova followed by Tukey-Kramer tests was used for statistical analyses (α = 0.05). RESULTS: Dentine treated with 1 or 500 mg ml(-1) of DAP followed by EDTA caused significant increases in DPSCs attachment compared to the dentine treated with the DAP alone. The 500 mg ml(-1) of DAP with or without EDTA caused significant reductions in DPSCs proliferation. However, the treatment of dentine with 1 mg ml(-1) of DAP did not have significant negative effects on DPSCs proliferation regardless of the use of EDTA. CONCLUSION: The use of 1 mg ml(-1) of DAP followed by 10 min of irrigation with EDTA in endodontic regeneration procedure may have no negative effects on the attachment and proliferation of DPSCs.

Evaluation of Residual Antibacterial Effect of Human Radicular Dentin Treated with Triple and Double Antibiotic Pastes.

INTRODUCTION: The aim of this study was to investigate the residual antibacterial effect of human radicular dentin treated with various concentrations of triple antibiotic paste and double antibiotic paste (DAP). METHODS: Sterilized dentin specimens were randomly assigned to 6 treatment groups and a no-treatment control group (n = 45 per group). For treatment groups, specimens were treated with either TAP or DAP at various concentrations (1000, 1, or 0.5 mg/mL) for 2 weeks. Then, each specimen was irrigated with 5 mL saline and incubated in phosphate-buffered solutions for 3, 7, 14, or 30 days. After that, Enterococcus faecalis was cultured on the specimens for 3 days. Each specimen was then transferred to a tube containing 200 µL saline, sonicated, and vortexed to detach the bacterial biofilm. The detached biofilm was spiral plated, and the number of colony-forming units was determined using an automated counting machine. RESULTS: Dentin specimens treated with 1000 mg/mL TAP or DAP had a significant residual antibacterial effect up to 14 days and 30 days, respectively. No significant difference was observed between 1000 and 1 mg/mL TAP and DAP at all time points. Dentin treated with all concentrations of DAP has a significantly longer residual antibacterial effect compared with dentin treated with TAP at the same concentrations. CONCLUSIONS: Radicular dentin treated with TAP and DAP showed a significant residual antibacterial effect compared with untreated dentin. All concentrations of DAP showed a significantly longer residual antibacterial effect compared with the same concentrations of TAP.

Effect of different endodontic regeneration protocols on wettability, roughness, and chemical composition of surface dentin.

INTRODUCTION: We investigated the changes in physiochemical properties of dentin surfaces after performing different endodontic regeneration protocols. METHODS: Human dentin slices were randomized into 4 treatment groups and 1 untreated control group (n = 10). One treatment group was irrigated with sodium hypochlorite (NaOCl) for 5 minutes followed by EDTA for 10 minutes. The other 3 treatment groups were irrigated with NaOCl; treated for 4 weeks with triple antibiotic paste (TAP), diluted triple antibiotic paste (DTAP), or calcium hydroxide (Ca[OH]2); and then irrigated with EDTA. After treatment, contact angles between a blood analog and dentin surfaces were evaluated. Surface roughness and chemical composition were characterized using optical profilometry and energy-dispersive X-ray spectroscopy, respectively. One-way analysis of variance followed by Fisher least significant difference tests were used for statistical analyses. RESULTS: All treatment groups showed a significant reduction in wettability and a significant increase in surface roughness when compared with untreated dentin. Dentin treated with Ca(OH)2 had significantly lower wettability compared with all other groups. No significant difference in wettability was found between dentin treated with DTAP and TAP protocols. Dentin treated with TAP had significantly higher surface roughness compared with all other groups. Untreated dentin and NaOCl + EDTA-treated dentin had significantly higher calcium and phosphorus as well as significantly lower carbon compared with dentin treated with Ca(OH)2, DTAP, and TAP. CONCLUSIONS: Endodontic regeneration protocols had a significant effect on wettability, surface roughness, and chemical composition of surface dentin. The Ca(OH)2 protocol caused a significant reduction in dentin wettability compared with TAP or DTAP protocols.

The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm.

OBJECTIVES: To investigate the effect of various dilutions of antibiotic medicaments used in endodontic regeneration on the survival of human dental pulp stem cells (DPSCs) and to determine their antibacterial effect against established Enterococcus faecalis biofilm. MATERIALS AND METHODS: The cytotoxic and antibacterial effects of different triple (TAP) and double antibiotic paste (DAP) dilutions (0.125, 0.25, 0.5, 1, and 10 mg/ml) were tested against Enterococcus faecalis established biofilm and DPSC. Established bacterial biofilm were exposed to antibiotic dilutions for 3 days. Then, biofilms were collected, spiral plated, and the numbers of bacterial colony forming units (CFU/ml) were determined. For the cytotoxic effect, lactate dehydrogenase activity assays (LDH) and cell viability assays (WST-1) were used to measure the percentage of DPSC cytotoxicity after 3-day treatment with the same antibiotic dilutions. A general linear mixed model was used for statistical analyses (α = 0.05). RESULTS: All antibiotic dilutions significantly decreased the bacterial CFU/ml. For WST-1 assays, all antibiotic dilutions except 0.125 mg/ml significantly reduced the viability of DPSC. For LDH assays, the three lowest tested concentrations of DAP (0.5, 0.25, 0.125 mg/ml) and the two lowest concentrations of TAP (0.25 and 0.125 mg/ml) were non-toxic to DPSC. CONCLUSIONS: All tested dilutions had an antibacterial effect against E. faecalis. However, 0.125 mg/ml of DAP and TAP showed a significant antibacterial effect with no cytotoxic effects on DPSCs. CLINICAL RELEVANCE: Using appropriate antibiotic concentrations of intracanal medicament during endodontic regeneration procedures is critical to disinfect root canal and decrease the adverse effects on stem cells.

A novel three-dimensional scaffold for regenerative endodontics: materials and biological characterizations.

An electrospun nanocomposite fibrous material holds promise as a scaffold, as well as a drug-delivery device to aid in root maturogenesis and the regeneration of the pulp-dentine complex. A novel three-dimensional (3D) nanocomposite scaffold composed of polydioxanone (PDS II®) and halloysite nanotubes (HNTs) was designed and fabricated by electrospinning. Morphology, structure, mechanical properties and cell compatibility studies were carried out to evaluate the effects of HNTs incorporation (0.5-10 wt% relative to PDS w/w). Overall, a 3D porous network was seen in the different fabricated electrospun scaffolds, regardless of the HNT content. The incorporation of HNTs at 10 wt% led to a significant (p < 0.0001) fibre diameter increase and a reduction in scaffold strength. Moreover, PDS-HNTs scaffolds supported the attachment and proliferation of human-derived pulp fibroblast cells. Quantitative proliferation assay performed with human dental pulp-derived cells as a function of nanotubes concentration indicated that the HNTs exhibit a high level of biocompatibility, rendering them good candidates for the potential encapsulation of distinct bioactive molecules. Collectively, the reported data support the conclusion that PDS-HNTs nanocomposite fibrous structures hold potential in the development of a bioactive scaffold for regenerative endodontics.

The use of traditional and novel techniques to determine the hardness and indentation properties of immature radicular dentin treated with antibiotic medicaments followed by ethylenediaminetetraacetic acid.

OBJECTIVE: The objective was to investigate the effect of intracanal antibiotic medicaments followed by ethylenediaminetetraacetic acid (EDTA) on the indentation properties and hardness of radicular dentin using a BioDent reference point indenter and a traditional microhardness technique, respectively. MATERIALS AND METHODS: Specimens with intact root canal dentin surfaces and polished radicular dentin specimens were obtained from immature human premolars. Each type of specimen was randomly assigned (n = 10 per group) and treated with either double antibiotic paste (DAP) for 4-week followed by EDTA for 5 min, triple antibiotic paste (TAP) for 4-week followed by EDTA for 5 min, EDTA for 5 min or Hank's balanced salt solution (control). The BioDent reference point indentor and Vickers microhardness tester were used to measure the indentation properties of root canal surfaces and the hardness of polished dentin specimens, respectively. One-way ANOVA followed by Fisher's protected least significant differences were used for statistical analyses. RESULTS: Both types of radicular dentin treated with antibiotic pastes and/or EDTA had a significant increase in the majority of indentation properties and a significant reduction in hardness compared to the untreated dentin. Furthermore, treatment of dentin with antibiotic pastes and EDTA caused significant increases in indentation properties and a significant reduction in hardness compared to EDTA-treated dentin. However, the RPI technique was not able to significantly differentiate between DAP + EDTA and TAP + EDTA-treated dentin. CONCLUSION: Dentin treated with antibiotic medicaments followed by EDTA had a significant increase the indentation properties and significantly reduction in hardness of radicular dentin.

Effects of two combinations of triple antibiotic paste used in endodontic regeneration on root microhardness and chemical structure of radicular dentine.

We investigated the effects of triple antibiotic paste (TAP) and modified triple antibiotic paste (MTAP) concentrations on the microhardness and chemical structure of radicular dentine. Human root cylinders were instrumented and randomized into four treatment groups and an untreated control group. Two treatment groups received 1 g/mL TAP or MTAP, and the other two treatment groups received 1 mg/mL methylcellulose-based TAP or MTAP. Cylinders were stored at 100% relative humidity for 4 weeks. Each root cylinder was subjected to a microhardness test before and after treatment. Different sets of radicular dentine specimens were treated as mentioned previously, and were examined using attenuated total reflection Fourier transform infrared spectroscopy. All treatment groups showed significant reductions in microhardness of roots when compared to untreated control roots at 1,000 and/or 500 µm from the pulp-dentine interface. However, 1 mg/mL methylcellulose-based antibiotics caused significantly less reduction in microhardness when compared to 1 g/mL antibiotics. In addition, 1 g/mL TAP and DAP caused significantly lower phosphate/amide I ratios when compared to other groups. The use of 1 mg/mL methylcellulose-based TAP and MTAP may minimize the reduction in microhardness of roots compared with the currently used 1 g/mL concentration of these antibiotics.

The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells.

AIM: The purpose of this in vitro study was to evaluate the effects of intracanal medicaments commonly used in endodontic regeneration on the survival of human dental pulp cells (DPCs). METHODS: DPCs were cultured and exposed to either no medicament treatment or low concentrations (0.3-5 mg ml(-1) ) of calcium hydroxide [Ca(OH)2 ], triple antibiotic paste (TAP), or double antibiotic paste (DAP) for 3 days. After that, toxicity to the DPCs was determined by lactate dehydrogenase activity assays (LDH) and cell proliferation was measured by colorimetric assays (WST-1). Two-way anova followed by Fisher's protected least significant differences was used for statistical analyses (α = 0.05). RESULTS: The group-by-concentration interactions were significant for the LDH and WST-1 assays (P < 0.0001). For the LDH assays, only the highest tested concentration (5 mg ml(-1) ) of Ca(OH)2 and TAP caused significant toxicity to the DPCs compared with the untreated control, while four tested concentrations of DAP (0.5, 1, 2.5, and 5 mg ml(-1) ) caused significant toxicity to the DPCs compared with the untreated control. For the WST-1 assays, the highest concentrations that did not negatively affect the proliferation rate of DAP, TAP and Ca(OH)2 were 0.3, 2, and 2.5 mg ml(-1) , respectively. CONCLUSION: The low concentrations of intracanal medicaments tested in this study were not cytotoxic in cultured cells. However, these concentrations are much lower than the concentrations that have been advocated in endodontic regeneration. Furthermore, the negative effects of TAP on DPCs were detected at lower concentrations by using the WST-1 assays than by measuring the LDH release.

A novel approach to evaluate the effect of medicaments used in endodontic regeneration on root canal surface indentation.

OBJECTIVES: The aim of this study was to investigate the capability of a novel reference point indentation apparatus to test the indentation properties of root canal surface dentine treated with three intracanal medicaments used in endodontic regeneration. MATERIALS AND METHODS: Immature human premolars were selected (n = 22). Four specimens were obtained from each root and randomly assigned to three treatment groups and a control group. Each specimen was exposed to one of the three treatment pastes (triple antibiotic (TAP), double antibiotic (DAP), or calcium hydroxide (Ca(OH)2)) or neutral deionized water (control) for 1 or 4 weeks. After each time interval, the indentation properties of the root canal dentine surfaces were measured using a BioDent reference point indenter. Two-way ANOVA and Fisher's protected least significant differences were used for statistical analyses. RESULTS: Significant differences in indentation parameters and estimated hardness between all groups at both time points were found. TAP-treated dentine had the highest significant indentation parameters, followed by DAP-treated dentine, untreated control dentine, and Ca(OH)2-treated dentine, respectively. Furthermore, TAP-treated dentine had the lowest significant estimated hardness, followed by DAP-treated dentine, untreated control dentine, and Ca(OH)2-treated dentine, respectively. CONCLUSION: BioDent reference point indenter was able to detect significant differences in indentation properties of root canal dentine treated with various medicaments. CLINICAL RELEVANCE: The use of a reference point indenter is a promising approach to characterize the indentation properties of root canal surfaces without any surface modification. This might provide an in vitro mechanical measurement that is more representative of the actual clinical situation.