Preliminary in vitro cytotoxic evaluation of Uncaria gambier (Hunt) Roxb extract as a potential herbal-based pulpotomy medicament.

BACKGROUND: The downfall of formocresol as a pulpotomy medicament highlights the importance of cytotoxic evaluation and the establishment of a safe concentration of dental material prior to its usage in the oral cavity. Uncaria gambir is an herbal plant that possesses antimicrobial, anti-inflammatory and antioxidant properties, suggesting its potential as an alternative medicament for pulpotomy. However, there are not many studies published on its cytotoxicity, with some using non-standardised techniques and reported variable outcomes. Here, we investigated the concentration and time-dependent toxicity of Uncaria gambir extract towards the M3CT3-E1 cell line and compared it with the gold standard pulpotomy medicament: mineral trioxide aggregate (MTA). METHODS: Uncaria gambir extracts at concentrations ranging from 1000 to 7.8 µg/ml and MTA eluates at 4- and 48 h setting times were prepared. 10% dimethyl sulfoxide (DMSO) and culture media were used as positive and negative controls respectively. Cell viability on days 1, 2, 3 and 7 was analysed using Alamar Blue and Live and Dead Cell assay. Any morphological cellular changes were evaluated using transmission electron microscopes (TEM). Data were analysed using a two-way mixed Analysis of Variance (ANOVA). RESULTS: The interaction between the concentration and exposure time on the fluorescence intensity of Uncaria gambir extract and MTA 48 h was found to be statistically significant (p < 0.001). No cytotoxic effects on the cells were exerted by both MTA 48 h and Uncaria gambir extract at a concentration below 500 µg/mL. TEM analysis and Live and Dead Cell assay for both materials were comparable to the negative control. No significant differences in fluorescent intensity were observed between Uncaria gambir extract at 500 µg/mL and MTA 48 h (p > 0.05). CONCLUSION: Uncaria gambir extracts at a maximum concentration of 500 µg/mL are non-cytotoxic over time and are comparable to the MTA.

The Combined Effects of Hydraulic Calcium Silicate Cement and Enamel Matrix Derivative Regarding Osteogenic and Dentinogenic Differentiation on Human Dental Pulp Stem Cells.

The ideal treatment option for immature necrotic permanent teeth is regeneration of the pulp-dentin complex. Mineral trioxide aggregate (MTA), the conventional cement used for regenerative endodontic procedures, induces hard tissue repair. Various hydraulic calcium silicate cements (HCSCs) and enamel matrix derivative (EMD) also promote osteoblast proliferation. The purpose of the present study was to determine the osteogenic and dentinogenic potential of commercially distributed MTA and HCSCs when applied in combination with Emdogain gel on human dental pulp stem cells (hDPSCs). The presence of Emdogain resulted in greater cell viability, and higher alkaline phosphatase activity was detected in the Emdogain-supplemented groups in the early days of cell culture. On qRT-PCR, the groups treated, respectively, with Biodentine and Endocem MTA Premixed in the presence of Emdogain showed an increased expression of the dentin formation marker DSPP, and the group treated with Endocem MTA Premixed in the presence of Emdogain showed an upregulated expression of the bone formation markers OSX and RUNX2. In an Alizarin Red-S staining assay, all of the experimental groups exhibited a greater formation of calcium nodules when treated in combination with Emdogain. Overall, the cytotoxicity and osteogenic/odontogenic potential of HCSCs were similar to that of ProRoot MTA. The addition of the EMD increased the osteogenic and dentinogenic differentiation markers.

Cytotoxic Effect of Nano Fast Cement and ProRoot Mineral Trioxide Aggregate on L-929 Fibroblast Cells: an in vitro Study.

Statement of the Problem: Endodontic materials that are placed in direct contact with living tissues should be biocompatible. The cytotoxicity of Nano Fast Cement (NFC) compared to ProRoot Mineral Trioxide Aggregate (ProRoot MTA) must be evaluated. Purpose: This In vitro study aimed to assess the cytotoxic effects of NFC in comparison to ProRoot MTA on L-929 mouse fibroblast cells. Materials and Method: In this animal study, L-929 mouse fibroblast cells were grown in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) in an atmosphere of 5% co2/95% air at 37 C̊. A total of 104 cells from the fourth collection were plated in each well of a 96-well micro-titer plate. Materials were mixed according to the manufacturer's instruction and placed into the related plastic molds with 5 mm diameter and 3 mm height. After 24 hours and a complete setting, the extracts of the tested materials were produced at six different concentrations and placed in the related wells. Cells in DMEM served as the negative control group. DMEM alone was used as the positive control group. Methyl-thiazoltetrazolium (MTT) colorimetric assay was conducted after 24, 48, and 72 hours. The absorbance values were measured by ELISA plate reader at 540 nm wavelength. Three-way analysis of variance, post-hoc Tukey, LSD, and independent t-test were used for the statistical analyses using SPSS software, version 16.0. Results: There was no statically significant difference between MTA and NFC in cell viability values at different concentrations and different time intervals (p= 0.649). Viability values were significantly decreased after 72 hours, but there was no significant difference between the first and second MTT assays (p= 0.987). Cytotoxicity significantly increased at concentrations higher than 6.25 µÉ¡/ml. Conclusion: Cytotoxicity depends on time, concentration, and cement composition. There was no statistically significant difference between NFC and MTA concerning their cytotoxic effects on L-929 mouse fibroblast cells.

Odonto/Osteogenic Differentiation of Dental Pulp Stem Cells of Type 1 Diabetic Patients with Mineral Trioxide Aggregate/1α,25-Dihydroxyvitamin D3 Combination.

INTRODUCTION: Vitamin D3 plays an important role in the mineralization mechanism and is often deficient in diabetic patients. The objective of this study was to investigate the odonto/osteogenic differentiation potential of the combination of mineral trioxide aggregate (MTA)/1α,25-dihydroxyvitamin D3 (VD3) on dental pulp stem cells (DPSCs) of patients with type 1 diabetes mellitus (T1DM). METHODS: DPSCs isolated from donors (control and T1DM) were cultured and characterized. Cell proliferation and wound healing assays were performed. DPSCs were exposed to 4 different media: growth medium (Dulbecco's modified Eagle's medium, 10% fetal bovine serum, antibiotic, and antimycotic), differentiation medium (DM) (growth medium plus ß-glycerophosphate and ascorbic acid), DM + MTA (DM plus 0.02 mg/mL MTA), and DM + MTA + VD3 (DM + MTA and 10 nmol/L vitamin D3). Odonto/osteogenic differentiation of DPSCs was evaluated by the alizarin red test, relative real-time polymerase chain reaction (dentin sialophosphoprotein, dentin matrix protein 1, collagen type 1 alpha 1, and osteocalcin), immunocytochemistry (antibone sialoprotein II, anti-dentin matrix protein 1, and anti-collagen type 1 alpha 1), and Western blot (dentin matrix protein 1 and osteocalcin) methods. RESULTS: The proliferation rates of DPSCs isolated from controls were significantly higher than DPSCs isolated from T1DM in a time-dependent manner (P < .05). Alizarin red staining and the expression of odonto/osteogenic markers showed that odonto/osteogenic differentiation was more pronounced in controls (P < .05) compared with T1DM patients. Although DM + MTA caused the odonto/osteogenic differentiation in DPSCs derived from controls, DM + MTA + VD3 resulted in the odonto/osteogenic differentiation in DPSCs of T1DM patients (P < .05). CONCLUSIONS: Odonto/osteogenic differentiation was affected by both supplements used for differentiation and the systemic disease, diabetes mellitus. The differentiation potential of T1DM-derived DPSCs was clearly increased with the VD3 supplement, although it was not as efficient as in the controls. The VD3 supplement showed a positive effect on the differentiation of T1DM DPSCs compared with MTA alone.

Effect of revascularisation and apexification procedures on biomechanical behaviour of immature maxillary central incisor teeth: a three-dimensional finite element analysis study.

OBJECTIVES: This study aimed to assess the effects of revascularisation and apexification procedures on biomechanical behaviour of immature teeth using 3-dimensional finite element analysis (3D FEA). MATERIALS AND METHODS: Five 3D FEA permanent maxillary incisor models were developed from CBCT scans and available literature data: Model MT: Mature tooth, Model IT: Immature tooth (Cvek's stage 3), Model AT: Apexified tooth-mineral trioxide aggregate (MTA) apexification, Model RTB: Revascularised tooth with blood, and Model RTS: Revascularised tooth with supplementary scaffold. Using FEA, a masticatory load of 240N at 120° was simulated, and the Von Mises and maximum principal stresses within the models were evaluated. Failure index (FI) and weakening% were also calculated for each model. RESULTS: On dentinal stress analysis, model MT (96.16MPa) and IT (158.38MPa) had lowest and highest stress values, respectively. Among the experimental groups, model RTS (131.12MPa) had lower stresses than AT (136.33MPa) and RTB (133.7MPa), with no significant difference among the three. Peak dentinal stresses in all the models were observed in the cervical third of the root and near the apical opening in model IT. The extent of high dentinal stress area in model RTB and RTS was lesser than that of AT. The FI and weakening% values were highest for model AT followed by RTB and RTS, among the experimental groups. However, all these treatments strengthened an immature tooth by more than 20%. CONCLUSIONS: AT, RTB, and RTS treatments lowered the stress values and risk of fracture in immature teeth with no significant difference among the three groups. CLINICAL RELEVANCE: Stress distribution evaluation following revascularisation/apexification was essential, with potential to influence clinical decision-making. MTA apexification and revascularisation with blood clot/supplementary scaffold lowered the stresses in immature teeth, with no significant difference among the three.

Comparative Evaluation of Mineral Trioxide Aggregate Pulpotomy and Laser-Assisted Mineral Trioxide Aggregate Pulpotomy: An Original Research Article.

CONTEXT AND AIM: Formocresol has been used as the material of choice (gold standard) for pulpotomy procedures because of the ease of use but was discouraged due to its potential immune sensitization and mutagenic effects. Laser irradiation was first applied for pulpotomy procedure in the year 1985. Recently, diode lasers have been used for pulpotomy in primary teeth and have shown clinical success rates comparable to formocresol. The present in vivo study was carried out to compare the clinical and radiographic success rates of mineral trioxide aggregate (MTA) pulpotomy and diode laser-assisted pulpotomy in human primary molars. MATERIALS AND METHODS: The present study was a randomized clinical trial in design wherein 40 primary teeth requiring pulpotomy treatment which met the selection criteria (clinical and radiographic) were divided into two groups, Group 1 (n = 20) wherein the pulpotomy was performed with MTA alone and Group 2 (n = 20) wherein laser-assisted pulpotomy was performed with MTA (L-MTA). The patients were recalled after 3, 6, and 9 months, respectively and evaluated clinically and radiographically. STATISTICAL ANALYSIS USED: The data were analyzed using the Statistical Package for the Social Sciences (SPSS) Version 22 (IBM corporation, Washington DC, United States). Descriptive statistics were used to analyze the data while the Pearson's correlation coefficient test was used to analyze the statistical correlation between the overall success rates observed in the clinical and radiographic findings of both the groups. P < 0.05 was considered statistically significant. RESULTS: The clinical success rate in the MTA group was 90%, 84.21%, and 88.23% at 3, 6, and 9 months, respectively, with no clinical signs or, symptoms reported at the said follow-up visits while the radiographic success rate was found to be 85%, 84.21%, and 82.3%, respectively. On the contrary, the clinical success rate in the L-MTA group was found to be 95%, 94.74%, and 94.44% at 3, 6, and 9, months, respectively with the radiographic success rate reported being 90%, 89.47%, and 88.89%, respectively. CONCLUSION: The combination of diode laser and MTA yielded better clinical and radiographic success rates over the pulpotomy procedures done with the help of MTA alone, thereby, concluding that lasers may be considered as adjuvant alternatives for vital pulp therapy on human primary teeth.

Raman spectroscopic study of the effect of the use of laser/LED phototherapy on the repair of complete tibial fracture treated with internal rigid fixation.

This study aimed to assess the repair of complete surgical tibial fractures fixed with internal rigid fixation (IRF) associated or not to the use of mineral trioxide aggregate (MTA) cement and treated or not with laser (λ = 780 nm, infrared) or LED (λ = 850 ±â€¯10 nm, infrared) lights, 142.8 J/cm2 per treatment, by means of Raman spectroscopy. Open surgical tibial fractures were created on 18 rabbits (6 groups of 3 animals per group, ∼8 months old) and fractures were fixed with IRF. Three groups were grafted with MTA. The groups of IRF and IRF + MTA that received laser or LED were irradiated every other day during 15 days. Animals were sacrificed after 30 days, being the tibia surgically removed. Raman spectra were collected via the probe at the defect site in five points, resulting in 15 spectra per group (90 spectra in the dataset). Spectra were collected at the same day to avoid changes in laser power and experimental setup. The ANOVA general linear model showed that the laser irradiation of tibial bone fractures fixed with IRF and grafted with MTA had significant influence in the content of phosphate (peak ∼960 cm-1) and carbonated (peak ∼1,070 cm-1) hydroxyapatites as well as collagen (peak 1,452 cm-1). Also, peaks of calcium carbonate (1,088 cm-1) were found in the groups grafted with MTA. Based on the Raman spectroscopic data collected in this study, MTA has been shown to improve the repair of complete tibial fractures treated with IRF, with an evident increase of collagen matrix synthesis, and development of a scaffold of hydroxyapatite-like calcium carbonate with subsequent deposition of phosphate hydroxyapatite.

Responses of oral-microflora-exposed dental pulp to capping with a triple antibiotic paste or calcium hydroxide cement in mouse molars.

INTRODUCTION: Responses of oral-microflora-exposed dental pulp to a triple antibiotic paste (TAP), a mixture of ciprofloxacin, metronidazole, and minocycline in ointment with macrogol and propylene glycol, remain to be fully clarified at the cellular level. This study aimed to elucidate responses of oral-microflora-exposed dental pulp to capping with TAP in mouse molars. METHODS: A cavity was prepared on the first molars of 6-week-old mice to expose the dental pulp for 24 h. The exposed pulp was capped with TAP (TAP group) or calcium hydroxide cement (CH group), in addition to the combination of macrogol (M) and propylene glycol (P) (MP, control group), followed by a glass ionomer cement filling. The samples were collected at intervals of 1, 2, and 3 weeks, and immunohistochemistry for nestin and Ki-67 and deoxyuride-5'-triphosphate biotin nick end labeling (TUNEL) assay were performed in addition to quantitative real-time polymerase chain reaction (qRT-PCR) analyses. RESULTS: The highest occurrence rate of pulp necrosis was found in the control group followed by the CH group at Weeks 2 and 3, whereas the highest occurrence rate of healed areas in the dental pulp was observed in the TAP group at each time point. Tertiary dentin formation was first observed in the dental pulp of the TAP group at Week 2. In contrast, bone-like and/or fibrous tissues were frequently observed in the CH group. qRT-PCR analyses clarified that TAP activated the stem and dendritic cells at Weeks 1 and 2, respectively. CONCLUSIONS: The use of TAP as a pulp-capping agent improved the healing process of oral-microflora-exposed dental pulp in mouse molars.

The Influence of New Silicate Cement Mineral Trioxide Aggregate (MTA Repair HP) on Metalloproteinase MMP-2 and MMP-9 Expression in Cultured THP-1 Macrophages.

The aim of the present study was to investigate the new silicate cement mineral trioxide aggregate (MTA Repair HP) with respect to its effect on the inflammation process involving the tooth and periodontal tissues. The composition of MTA Repair HP was supplemented with plasticizer agents which can have a negative effect on the modulation of tooth inflammation. The silicate-based material in question is widely used in regeneration of the pulp-dentin complex, treatment of perforations of various locations in the tooth, as well as in surgical treatment of the complications of periapical tissue. The improved bioceramic restorative cement can affect the expression of metalloproteinases MMP-2 and MMP-9 in monocytes/macrophages involved in modulation of inflammation and regenerative processes of the tooth and periodontal tissues. The novel aspect of the present study lies in the application of the model of THP-1 monocyte/macrophage and applying the biomaterial in direct contact with the cells. Hence, it provides a representation of clinical conditions with respect to regenerative pulp and periodontal treatment with the use of MTA Repair HP. A lack of macrophage activation (as measured with flow cytometry) was found. Moreover, the study identified a lack of expression stimulation of the studied metalloproteinases (with the use of Western blotting and fluorescent microscopy). Similarly, no increase in MMP-2 and MMP-9 concentration was found (measured by ELISA method) in vitro when incubated with MTA Repair HP. Based on the results it can be concluded that new MTA Repair HP does not increase the inflammatory response in monocytes/macrophages associated with the activity of the described enzymes. It can also be speculated that they do not affect the process of dentin regeneration in which MMP-2 and MMP-9 play significant roles.

In vitro evaluation of cytotoxicity of Emblica officinalis (amla) on cultured human primary dental pulp fibroblasts.

CONTEXT: The dental pulp tissue is capable of healing after surgical amputation of infected/inflamed tissue during vital pulp therapy, when in contact with a suitable medicament. Emblica officinalis (amla), a traditional medicine, is one such medicament which has never been evaluated for its healing potential in pulp therapy. AIMS: The aim of the study was to evaluate the cytotoxicity of E. officinalis (amla) against human primary dental pulp fibroblasts. SETTINGS AND DESIGN: This was in vitro study. SUBJECTS AND METHODS: Human dental pulp fibroblasts were obtained from dental pulp tissue of extracted over-retained primary incisors. The primary cells were cultured using the Dulbecco's Modified Eagle's Medium and used for the study after the fourth passage. The test medicament was E. officinalis with mineral trioxide aggregate (MTA) (100%) and untreated cells as positive and negative controls, respectively. Methyl-thiazol-diphenyl-tetrazolium (MTT) cytotoxicity assay was performed, and the cell survival was observed and analyzed at intervals of 24, 48, and 72 h. STATISTICAL ANALYSIS USED: Cell survival within groups was compared with Wilcoxon matched-paired t-test and in between groups at each point interval was analyzed with the Kruskal-Wallis ANOVA test. The level of significance was set at 0.05. RESULTS: Within the groups, across the time periods of evaluation, there was a decline in cell survival in both the groups but was statistically significant in the MTA group. On interval-wise comparison, the decline in cell survival was statistically significant between the three groups at 72 h (P = 0.001). CONCLUSIONS: E. officinalis preserved the vitality of the human primary dental pulp fibroblasts and has the potential to be developed into vital pulp therapy medicament.

Identification of a Calcium-sensing Receptor in Human Dental Pulp Cells That Regulates Mineral Trioxide Aggregate-induced Mineralization.

INTRODUCTION: The purpose of this study was to verify the expression of the calcium-sensing receptor (CaSR) and its role in mineral trioxide aggregate (MTA)-induced odontoblastic differentiation and mineralization in human dental pulp cells (hDPCs). METHODS: The expression of CaSR in human dental pulp tissue and hDPCs was detected using immunohistochemical and immunofluorescent assays. Then, hDPCs were cultured in specific medium supplemented with defined concentrations of MTA dilute alone or in combination with calcimimetic R-568 (a positive allosteric modulator of CaSR [Tocris Bioscience, Bristol, UK]), and cell viability was monitored by Cell Counting Kit-8 (Dojindo Molecular Technologies, Kumamoto, Japan) analysis. Alkaline phosphatase activity, alizarin red S staining, quantitative real-time polymerase chain reaction, and Western blot were used to investigate the gene/protein expression of odontoblastic-associated markers and CaSR in medium supplemented with different combinations of diluted MTA, R-568, and calcilytic Calhex 231 (a negative allosteric modulator of CaSR [Sigma-Aldrich, St Louis, MO]). RESULTS: CaSR was slightly expressed in the central pulp tissue, whereas it was strongly expressed in the odontoblast layer, plasma membrane, and cytoplasm of hDPCs. Cell Counting Kit-8 assay indicated maximum cell viability in cultures treated with 1:8 diluted MTA additives. Compared with undifferentiated controls, the cells at the early stage of odontoblastic differentiation exhibited lower CaSR protein expression. The combination of 1:8 diluted MTA with 0.1 and 1.0 µmol/L R-568 led to significantly increased cell vitality but decreased alkaline phosphatase activity and mineralized deposit formation, and this negative effect could be attenuated by 1.0 µmol/L Calhex 231 supplementation. Quantitative polymerase chain reaction results showed a significant up-regulation of RUNX2, DSPP, DMP-1, and OCN gene expression in the 1 µmol/L R-568-treated hDPCs. Western blot analysis indicated that the treatment by MTA and R-568 alone or their combination gave no clear trend on the protein levels of CaSR and dentin sialophosphoprotein, whereas Calhex 231 can increase their expressions. In addition, the up-regulation of Akt phosphorylation was observed in R-568- and Calhex 231-treated hDPCs. CONCLUSIONS: Our data indicated that CaSR is expressed in human dental pulp and hDPCs and that it can negatively or positively regulate MTA-induced mineralization of hDPCs via the phosphoinositide 3-kinase/Akt pathway in a ligand-dependent manner, suggesting a therapeutic target for modulating reparative dentin formation.

Antimicrobial effect of bioceramic cements on multispecies microcosm biofilm: a confocal laser microscopy study.

OBJECTIVES: To assess the viability of multispecies microcosm biofilm after contact with NeoMTA Plus, Biodentine, and MTA Angelus. MATERIALS AND METHODS: Fifty-four human dentin blocks (4 × 5 × 4 mm) were allocated to Hawley retainers, worn by six volunteers for 72 h. The blocks were then individually incubated in BHI broth for 21 days at 37 °C. At the end of experimental time for biofilm growth, the samples were randomly divided into four groups (n = 12): NeoMTA Plus, Biodentine, MTA Angelus, and negative control. The materials were placed in contact with the blocks. All samples were placed in cell-culture plate wells and incubated in BHI broth for 7 days at 37 °C. One sample from each volunteer (n = 6) was analyzed by SEM to describe the biofilm morphology. CLSM was performed to determine the percentage of viable biofilm biovolume. The data were statistically analyzed by one-way ANOVA and Tukey's multiple comparison test (α = 5%). RESULTS: SEM showed biofilm formed by spherical and rod-shaped bacteria surrounded by an extracellular matrix. No material was able to kill all biofilm cells, and all groups had more than 50% of viable bacteria. NeoMTA Plus was significantly different from the negative control group (P < .05). CONCLUSIONS: All tested materials were not effective against multispecies microcosm biofilm. CLINICAL RELEVANCE: NeoMTA Plus, Biodentine, and MTA Angelus were not effective against multispecies microcosm biofilm. It is essential to understand that these bioceramic cements are indicated for infected clinical situations. Thus, complementary disinfection procedures should be conducted prior to filling with these materials.

Clinical and histological responses of human dental pulp to MTA and combined MTA/treated dentin matrix in partial pulpotomy.

The aim of this study was to compare the responses of mineral trioxide aggregate (MTA) and combined MTA/treated dentin matrix (TDM) as direct pulp capping material. In this clinical trial study, 33 intact third molars in 11 healthy volunteers (three molars in each) were included. Partial pulpotomies were performed in a split mouth manner in two of the third molars in each patient randomly and the third tooth had used as TDM source. The coronal dentin was chopped to dentine chips and transformed to TDM using different concentrations of ethylene diamine tetraacetic acid (EDTA) solution. Pulps were directly capped by MTA alone or using a combination layering of MTA/TDM. Following 6-week clinical and radiological evaluations, each tooth was extracted and prepared for histological evaluation. There were no significant differences in the clinical and radiographic responses or in the quality of dentin bridges (P > 0.05). However, the dentin bridge was significantly thicker in MTA/TDM group than MTA group (P = 0.003). Using the combination of MTA/TDM as a pulp-dressing agent may form a thicker dentin bridge compared to MTA alone.

The Effect of Three Different Biomaterials on Proliferation and Viability of Human Dental Pulp Stem Cells (In-vitro Study).

BACKGROUND: Biomaterial cytotoxicity on dental stem cells plays a critical role in managing the regeneration of dental tissue. AIM: The aim of the present study was to evaluate the effect of Nano-hydroxy apatite (NHA), Mineral trioxide aggregate (MTA), and Calcium-enriched mixture (CEM) on the proliferation, and viability of human dental pulp stem cells (hDPSCs) isolated from third molar teeth. METHODS: Cultured DPSCs were characterized and the tested biomaterials were shaped into cylinders then inserted directly on the DPSCs. Proliferation and viability percentage of DPSCs were evaluated at 1, 3, 5, 7, 9, 11, and 14 days of culture. RESULTS: The biomaterials supplemented DPSCs showed a significant initial decrease in cell count and viability percentage at day one. Then, a rise in cell counts and viabilities was noticed after that. There was a decrease in cell counts, and viabilities in the NHA supplemented cells in comparison to other tested biomaterials. CONCLUSIONS: All tested biomaterials maintain the proliferation of DPSCs for different durations. NHA showed less proliferative and more cytotoxic effect than other tested materials.

Antimicrobial and antibiofilm activities of MTA supplemented with bismuth lipophilic nanoparticles.

The objective of this work was to determine the antimicrobial and antibiofilm properties of mineral trioxide aggregate (MTA) supplemented with bismuth lipophilic nanoparticles (BisBAL NPs). The antimicrobial activity of the composite MTA-BisBAL NPs was determined by the disk diffusion assay, while antibiofilm activity was analyzed by fluorescence microscopy. The cytotoxicity of MTA-BisBAL NPs was determined on human gingival fibroblasts by optical microscopy and crystal violet staining. MTA-BisBAL NPs inhibited the growth of Enterococcus faecalis, Escherichia coli, and Candida albicans and also detached the biofilm of fluorescent E. faecalis after 24 h of treatment. The addition of BisBAL nanoparticles did not significantly modify the physical properties of MTA, and cytotoxicity was not observed when MTA-BisBAL NPs was added on human gingival fibroblasts. Altogether these results suggest that BisBAL nanoparticles provide antimicrobial and antibiofilm activities to MTA while it retained their biophysical properties without cause side effects on human gingival fibroblasts.

Outcome of Revascularization Procedure: A Retrospective Case Series.

INTRODUCTION: The purpose of this retrospective case series was to investigate the outcome of the revascularization procedure in necrotic immature teeth. METHODS: The residents and faculty members at the University of Pennsylvania endodontic department were invited to submit consecutive revascularization cases treated by them, irrespective of the outcome, during the time period of 2009 to 2012. Twenty-eight of 35 submitted necrotic immature teeth met the inclusion criteria. The treatment protocol included minimal instrumentation and irrigation with 3% sodium hypochlorite and 17% EDTA. Triple antibiotic paste was placed for a minimum of 21 days. After blood clot induction, either EndoSequence Bioceramic Putty (Brasseler, Savannah, GA) or mineral trioxide aggregate was placed below the cementoenamel junction, and composite was used as a final restoration. The follow-up period ranged from 7 to 72 months. The outcome was assessed as complete healing (the absence of clinical signs and symptoms, complete resolution of periradicular radiolucency, increase in the root dentin thickness/length, and apical closure), incomplete healing (the absence of clinical signs and symptoms, the periapical lesion completely healed without any signs of root maturation or thickening, the periapical lesion either reduced in size or unchanged with/without radiographic signs of increasing root dentin thickness/length, or apical closure), and failure (persistent clinical signs and symptoms and/or increased size of the periradicular lesion). RESULTS: Twenty-one of 28 cases (75%) healed completely, 3 cases (10.7%) failed during the observation period and needed further treatment, and 4 cases (14%) presented with incomplete healing. CONCLUSIONS: Within the limitation of this study, the outcome of revascularization, wherein healing of periapical periodontitis and maturation of roots occurs, is fairly high, making it a viable treatment option in comparison with apexification.

Effect of the plant-based hemostatic agent Ankaferd Blood Stopper® on the biocompatibility of mineral trioxide aggregate.

BACKGROUND: Due to the detrimental effect of blood contamination on the physico-chemical properties of mineral trioxide aggregate (MTA), obtaining an effective hemostasis in the surgical crypt during apical surgery is of paramount importance. The purpose of this in vivo study was to analyze the effect of Ankaferd Blood Stopper® (ABS) contamination on the biocompatibility of MTA. METHODS: Forty of 56 Wistar-Albino rats were divided randomly and equally into two groups (MTA and MTA-ABS) according to whether or not a hemostatic agent was used. The remaining 16 rats were designated as the control group. Rats in the experimental groups received freshly mixed MTA-Angelus in polyethylene tubes, which were inserted into monocortical bore holes created in their tibias. In the MTA-ABS group only, 0.5 mL of ABS solution was administered topically on the defect sites followed by implantation of MTA tubes. Inflammation, foreign-body reaction (FBR), necrosis, fibrosis, and new bone formation (NBF) were studied 7, 30, 60, and 90 days after implantation. RESULTS: On day7, statistically significant differences were found in tissue reactions with regard to NBF and necrosis (p = 0.044 and p = 0.024, respectively), the latter being observed in 40 % of the samples only in the MTA-ABS group. Slight inflammation in all groups was confined to day-7 only. Mild necrosis was present in the MTA-ABS group only on day-7. Severity of the foreign body reaction and fibrosis was limited. New bone formation increased gradually over time in all groups, reaching a maximum on day-90. CONCLUSIONS: MTA and ABS-contaminated MTA are equally biocompatible. ABS does not impair the properties of MTA.

Interfacial characteristics of Biodentine and MTA with dentine in simulated body fluid.

OBJECTIVES: Newer tricalcium silicate cements (TSC) may offer biocompatibility with improved working properties. This study aimed to evaluate: (1) the occurrence of mineral deposition at the interface between dentine and two TSC (ProRoot(®) MTA and Biodentine(®)) in simulated body fluid, and (2) to investigate the nature of interfacial layer. METHODS: Six root dentine segments of 1.5mm thickness were obtained from extracted human teeth and were instrumented with Gates-Glidden drills. The specimens were then randomly filled with either MTA or Biodentine. The specimens were placed in the simulated body fluid containing the same phosphate concentration as blood plasma. After 4 weeks, the specimens were examined with Scanning Electron Microscope (SEM) and Energy Disperse X-ray Spectroscopy (EDX) to measure the thickness of the interfacial layer and Ca/P ratio. Transmission Electron Microscope (TEM) and Selective Area Electron Diffraction (SAED) were conducted to examine the interface ultramicroscopically and to determine the nature of the crystalline structure within interfacial layer. RESULTS: The thickness of interfacial layer was significantly higher in the MTA group (14.5 µm vs 4.8 µm) (p<0.001). However, there was no significant difference between MTA and Biodentine in Ca/P ratio of interfacial layer (4.1 vs 2.7) (p>0.05). From TEM examination, amorphous calcium phosphate (ACP) was observed in the interface along with the surface of dentine. CONCLUSIONS: As an alternative to MTA, Biodentine displayed bioactivity by producing an interfacial layer on the root canal dentine even though its thickness was significantly lower than MTA. ACP was observed in the interfacial layer of both biomaterials. CLINICAL SIGNIFICANCE: Biodentine could be considered as an alternative to MTA due to comparable bioactivity which creates interfacial layer between root canal dentin and Biodentine.

Regenerative endodontic therapy: a data analysis of clinical protocols.

INTRODUCTION: The aim of the present study was to systematically analyze the protocols that have been used in regenerative endodontic therapy and to detect any variations in clinical procedures. METHODS: An electronic search was executed in PubMed using appropriate Medical Subject Heading terms covering the period from January 1993 to May 2014. Additional publications from hand searching and the reference section of each relevant article enriched the article list. The relevance of each article was initially evaluated by scanning all titles and corresponding abstracts. The definite inclusion of each article in the study was determined by using specific criteria applied independently by 3 reviewers. RESULTS: Sixty relevant publications were finally included. The canal walls were not mechanically instrumented in 68% of the clinical articles. Sodium hypochlorite was included in 97% of the clinical studies either as the only irrigant or in combination with other irrigants. Antibiotic combination paste was used as the intracanal medicament in 80% of the clinical articles. Sodium hypochlorite, chlorhexidine, and EDTA were used in the final irrigation protocol in 75%, 4%, and 13% of the clinical studies, respectively. Neither the creation of a blood clot nor the use of platelet-rich plasma/platelet-rich fibrin was described in 13% of the clinical articles. Mineral trioxide aggregate was used as an intracanal coronal barrier in 85% of the relevant clinical studies. CONCLUSIONS: The variability of the clinical protocols applied during regenerative enododontic procedures is considerably high. A thorough analysis of regenerative protocols may constitute an additional source to provide useful clinical considerations for REPs.

Capping a pulpotomy with calcium aluminosilicate cement: comparison to mineral trioxide aggregates.

INTRODUCTION: Calcium aluminate cements have shown little affinity for bacterial growth, low toxicity, and immunogenicity when used as a restoration material, but calcium aluminate cements have not been tested in vivo in pulpotomy procedures. METHODS: To address this question, a calcium aluminosilicate cement (Quick-Set) was tested along with 2 mineral trioxide aggregates, ProRoot MTA and MTA Plus. These cements were used as a capping agent after pulpotomy. Control rats had no pulpotomy, or the pulpotomy was not capped. Proinflammatory cytokines interleukin (IL)-1ß and IL-1α were measured, and histology was performed at 30 and 60 days after capping. The nociceptive response was determined by measuring the lengthening of the rat's meal duration. RESULTS: and CONCLUSIONS: IL-1ß and IL-1α concentrations were reduced in the capped teeth, but no differences were observed among the 3 cements. Dentinal bridging could be detected at both 30 and 60 days with each of the 3 cements, and the pulps were still vital 60 days after capping. Meal duration significantly shortened after placement of the 3 different cements, indicating a nociceptive response, but there were no differences among the materials. Calcium aluminosilicate cement had similar properties to mineral trioxide aggregates and is a viable option for pulpotomy procedures.