Differentiation of human endometrial stem cells encapsulated in alginate hydrogel into oocyte-like cells.

Introduction: Human endometrial mesenchymal stem cells (hEnMSCs) are a rich source of mesenchymal stem cells (MSCs) with multi-lineage differentiation potential, making them an intriguing tool in regenerative medicine, particularly for the treatment of reproductive and infertility issues. The specific process of germline cell-derived stem cell differentiation remains unknown, the aim is to study novel ways to achieve an effective differentiation method that produces adequate and functioning human gamete cells. Methods: We adjusted the optimum retinoic acid (RA) concentration for enhancement of germ cell-derived hEnSCs generation in 2D cell culture after 7 days in this study. Subsequently, we developed a suitable oocyte-like cell induction media including RA and bone morphogenetic protein 4 (BMP4), and studied their effects on oocyte-like cell differentiation in 2D and 3D cell culture media utilizing cells encapsulated in alginate hydrogel. Results: Our results from microscopy analysis, real-time PCR, and immunofluorescence tests revealed that 10 µM RA concentration was the optimal dose for inducing germ-like cells after 7 days. We examined the alginate hydrogel structural characteristics and integrity by rheology analysis and SEM microscope. We also demonstrated encapsulated cell viability and adhesion in the manufactured hydrogel. We propose that in 3D cell cultures in alginate hydrogel, an induction medium containing 10 µM RA and 50 ng/mL BMP4 can enhance hEnSC differentiation into oocyte-like cells. Conclusion: The production of oocyte-like cells using 3D alginate hydrogel may be viable in vitro approach for replacing gonad tissues and cells.

MicroRNAs-mediated regulation of the differentiation of dental pulp-derived mesenchymal stem cells: a systematic review and bioinformatic analysis.

BACKGROUND: Human dental pulp-derived mesenchymal stem cells (hDP-MSCs), which include human dental pulp stem cells (hDPSCs) and stem cells from human exfoliated deciduous teeth (SHEDs), are promising cell sources for regenerative therapies. Nevertheless, a lack of knowledge relating to the mechanisms regulating their differentiation has limited their clinical application. microRNAs (miRNAs) are important regulatory molecules in cellular processes including cell differentiation. This systematic review aims to provide a panel of miRNAs that regulate the differentiation of hDP-MSCs including hDPSCs and SHEDs. Additionally, bioinformatic analyses were conducted to discover target genes, signaling pathways and gene ontologies associated with the identified miRNAs. METHODS: A literature search was performed in MEDLINE (via PubMed), Web of Science, Scopus, Embase and Cochrane Library. Experimental studies assessing the promotive/suppressive effect of miRNAs on the differentiation of hDP-MSCs and studies evaluating changes to the expression of miRNAs during the differentiation of hDP-MSCs were included. miRNAs involved in odontogenic/osteogenic differentiation were then included in a bioinformatic analysis. A miRNA-mRNA network was constructed, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. A protein-protein interaction (PPI) network was also constructed. RESULTS: Of 766 initially identified records through database searching, 42 and 36 studies were included in qualitative synthesis and bioinformatic analyses, respectively. Thirteen miRNAs promoted and 17 suppressed odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-140-5p, hsa-miR-218 and hsa-miR-143 were more frequently reported suppressing the odontogenic/osteogenic differentiation of hDP-MSCs. hsa-miR-221 and hsa-miR-124 promoted and hsa-miR-140-5p inhibited neuronal differentiation, hsa-miR-26a-5p promoted and hsa-miR-424 suppressed angiogenic differentiation, and hsa-miR-135 and hsa-miR-143 inhibited differentiation within myogenic lineages. A miRNA-mRNA network including 1890 nodes and 2171 edges was constructed. KEGG pathway analysis revealed MAPK, PI3K-Akt and FoxO as key signaling pathways involved in the odontogenic/osteogenic differentiation of hDP-MSCs. CONCLUSIONS: The findings of this systematic review support the potential application of the specific miRNAs to regulate the directed differentiation of hDP-MSCs in the field of regenerative therapies.

Preparation and In Vitro Osteogenic Evaluation of Biomimetic Hybrid Nanocomposite Scaffolds Based on Gelatin/Plasma Rich in Growth Factors (PRGF) and Lithium-Doped 45s5 Bioactive Glass Nanoparticles.

Bone tissue engineering is an emerging technique for repairing large bone lesions. Biomimetic techniques expand the use of organic-inorganic spongy-like nanocomposite scaffolds and platelet concentrates. In this study, a biomimetic nanocomposite scaffold was prepared using lithium-doped bioactive-glass nanoparticles and gelatin/PRGF. First, sol-gel method was used to prepare bioactive-glass nanoparticles that contain 0, 1, 3, and 5%wt lithium. The lithium content was then optimized based on antibacterial and MTT testing. By freeze-drying, hybrid scaffolds comprising 5, 10, and 20% bioglass were made. On the scaffolds, human endometrial stem cells (hEnSCs) were cultured for adhesion (SEM), survival, and osteogenic differentiation. Alkaline phosphatase activity and osteopontin, osteocalcin, and Runx2 gene expression were measured. The effect of bioactive-glass nanoparticles and PRGF on nanocomposites' mechanical characteristics and glass-transition temperature (T g) was also studied. An optimal lithium content in bioactive glass structure was found to be 3% wt. Nanoparticle SEM examination indicated grain deformation due to different sizes of lithium and sodium ions. Results showed up to 10% wt bioactive-glass and PRGF increased survival and cell adhesion. Also, Hybrid scaffolds revealed higher ALP-activity and OP, OC, and Runx2 gene expression. Furthermore, bioactive-glass has mainly increased ALP-activity and Runx2 expression, whereas PRGF increases the expression of OP and OC genes. Bioactive-glass increases scaffold modulus and T g continuously. Hence, the presence of both bioactive-glass and nanocomposite scaffold improves the expression of osteogenic differentiation biomarkers. Subsequently, it seems that hybrid scaffolds based on biopolymers, Li-doped bioactive-glass, and platelet extracts can be a good strategy for bone repair.

Antibacterial efficacy of antibiotic pastes versus calcium hydroxide intracanal dressing: A systematic review and meta-analysis of ex vivo studies.

Background: Conflicting findings on the potency of antibiotic pastes versus calcium hydroxide (CH) have been evident in the literature. Aims: To compare the antibacterial efficacy of single antibiotic paste (SAP), double antibiotic paste (DAP), triple antibiotic paste (TAP), and modified TAP (mTAP) with CH on bacterial biofilms. Methods: PubMed, Scopus, and Embase were comprehensively searched until August 23, 2021. The study protocol was registered in the PROSPERO. Ex vivo studies performed on Enterococcus faecalis or polymicrobial biofilms incubated on human/bovine dentin were selected. The quality of the studies was assessed using a customized quality assessment tool. Standardized mean difference (SMD) with a 95% confidence interval (CI) was calculated for the meta-analysis. Meta-regression models were used to identify the sources of heterogeneity and to compare the efficacy of pastes. Results: The qualitative and quantitative synthesis included 40 and 23 papers, respectively, out of 1421 search results. TAP (SMD = -3.82; CI, -5.44 to -2.21; P < 0.001) and SAPs (SMD = -2.38; CI, -2.81 to - 1.94; P < 0.001) had significantly higher antibacterial efficacy compared to the CH on E. faecalis biofilm. However, no significant difference was found between the efficacy of DAP (SMD = -2.74; CI, -5.56-0.07; P = 0.06) or mTAP (SMD = -0.28; CI, -0.82-0.26; P = 0.31) and CH. Meta-regression model on E. faecalis showed that SAPs have similar efficacy compared to TAP and significantly better efficacy than DAP. On dual-species (SMD = 0.15; CI, -1.00-1.29; P = 0.80) or multi-species (SMD = 0.23; CI, -0.08-0.55; P = 0.15) biofilms, DAP and CH had similar efficacy. Conclusions: Ex vivo evidence showed that antibiotic pastes were either superior or equal to CH. The studied SAPs had considerably higher or similar antibacterial effectiveness compared to DAP, CH, and TAP. Hence, combined antibiotic therapy was not necessarily required for root canal disinfection ex vivo.

Comparative evaluation of the effects of three hydraulic calcium silicate cements on odontoblastic differentiation of human dental pulp stem cells: an in vitro study.

OBJECTIVE: The study aimed to compare the response of human dental pulp stem cells (hDPSCs) towards three hydraulic calcium silicate cements (HCSCs) by measuring cytotoxicity and expression of dentinogenic genes. METHODOLOGY: Dental pulps of five impacted mandibular third molars were extirpated as a source for hDPSCs. Next to culturing, hDPSCs were subjected to fluorescence-activated cell sorting after the third passage to validate stemness of the cells. Human DPSCs were exposed to diluted supernatants of OrthoMTA (OMTA), Biodentine (BD) and Calcium-Enriched Mixture (CEM) at concentrations 10, 25, 50 and 100% at the first, third and fifth day of culture. Then, cells were exposed to 10% concentrations supernatant of HCSCs to determine DSPP and DMP1 gene expression, using a quantitative polymerase-chain reaction. Data were analyzed using one-way and three-way ANOVA, followed by Tukey post hoc statistical tests. RESULTS: Optimal cell proliferation was observed in all groups, regardless of concentration and time-point. HCSC supernatants were non-cytotoxic to hDPSCs at all three time-points, except for 100% Biodentine on day five. On day seven, OMTA group significantly upregulated the expression of DSPP and DMP1 genes. On day 14, expression of DMP1 and DSPP genes were significantly higher in BD and OMTA groups, respectively. CONCLUSION: Biodentine significantly upregulated DMP1 gene expression over 14 days, whereas CEM was associated with only minimal expression of DSPP and DMP1 .

Comparative evaluation of the effects of three hydraulic calcium silicate cements on odontoblastic differentiation of human dental pulp stem cells: an in vitro study

Abstract Objective The study aimed to compare the response of human dental pulp stem cells (hDPSCs) towards three hydraulic calcium silicate cements (HCSCs) by measuring cytotoxicity and expression of dentinogenic genes. Methodology Dental pulps of five impacted mandibular third molars were extirpated as a source for hDPSCs. Next to culturing, hDPSCs were subjected to fluorescence-activated cell sorting after the third passage to validate stemness of the cells. Human DPSCs were exposed to diluted supernatants of OrthoMTA (OMTA), Biodentine (BD) and Calcium-Enriched Mixture (CEM) at concentrations 10, 25, 50 and 100% at the first, third and fifth day of culture. Then, cells were exposed to 10% concentrations supernatant of HCSCs to determine DSPP and DMP1 gene expression, using a quantitative polymerase-chain reaction. Data were analyzed using one-way and three-way ANOVA, followed by Tukey post hoc statistical tests. Results Optimal cell proliferation was observed in all groups, regardless of concentration and time-point. HCSC supernatants were non-cytotoxic to hDPSCs at all three time-points, except for 100% Biodentine on day five. On day seven, OMTA group significantly upregulated the expression of DSPP and DMP1 genes. On day 14, expression of DMP1 and DSPP genes were significantly higher in BD and OMTA groups, respectively. Conclusion Biodentine significantly upregulated DMP1 gene expression over 14 days, whereas CEM was associated with only minimal expression of DSPP and DMP1 .

Epinephrine-entrapped chitosan nanoparticles covered by gelatin nanofibers: A bi-layer nano-biomaterial for rapid hemostasis.

Uncontrolled hemorrhage accounts for significant death risk both in trauma and surgery. Various bleeding control techniques have been emerged to augment hemostasis, which still has several limitations and drawbacks. In this study, epinephrine-entrapped chitosan nanoparticles were electrosprayed on a base pad and covered by a gelatin nanofiber layer (E-CS-Gl. Physico-chemical characteristics, hemocompatibility, cytotoxicity, and blood coagulation tests were studied in-vitro, and blood coagulation and hemostasis potential tests were performed in-vivo. The in-vitro results showed that the prepared nano-biomaterial is cytocompatible against HuGu cells. Also, hemocompatibility studies showed that PT and aPTT times did not change in comparison with the controls. Further blood coagulation study indicated that E-CS-Gl provides an ultimate interface to induce red blood cell absorption and aggregation, resulting in augmented blood coagulation. E-CS-Gl also caused rapid clotting in rat models of ruptured femoral artery and liver compared to controls. Findings exhibited that E-CS-Gl is a safe and effective hemostatic agent and provides a new approach for fast and safe hemorrhage control.

Isolation and Differentiation of Adipose-Derived Stem Cells into Odontoblast-Like Cells: A Preliminary In Vitro Study.

OBJECTIVE: The aim of present study was to isolate and differentiate human adipose-derived stem cells (ASCs) into odontoblast-like cells. MATERIALS AND METHODS: In this experimental study, human adipose tissues were taken from the buccal fat pad of three individuals (mean age: 24.6 ± 2.1 years). The tissues were transferred to a laboratory in a sterile culture medium, divided into small pieces and digested by collagenase I (2 mg/mL, 60-90 minutes). ASCs were isolated by passing the cell suspension through cell strainers (70 and 40 µm), followed by incubation at 37ºC and 5% CO2 in Dulbecco's modified eagle medium (DMEM) supplemented with fetal bovine serum (FBS 5%) and penicillin/streptomycin (P/S). After three passages, the ASCs were harvested. Subsequently, flow cytometry and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect expression levels of NANOG and OCT4 to evaluate stemness. Then, a differentiation medium that included high-glucose DMEM supplemented with 10% FBS, dexamethasone (10 nM), sodium ß-glycerophosphate (5 mM) and ascorbic acid (100 µM) was added. The cells were cultivated for four weeks, and the odontogenic medium was changed every two days. Cell differentiation was evaluated with Alizarin red staining and expressions of collagen I (COL1A1), dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP1). RESULTS: The ASCs were effectively and easily isolated. They were negative for CD45 and positive for the CD105 and CD73 markers. The ASCs expressed OCT4 and NANOG. Differentiated cells highly expressed DSPP, COL1A1 and DMP1. Alizarin red staining revealed a positive reaction for calcium deposition. CONCLUSION: ASCs were isolated successfully in high numbers from the buccal fat pad of human volunteers and were differentiated into odontoblast-like cells. These ASCs could be considered a new source of cells for use in regenerative endodontic treatments.

Application of Platelet Rich Fibrin in Tissue Engineering: Focus on Bone Regeneration.

Bone tissue engineering (BTE) is a strategy for reconstructing bone lesions, which is rapidly developing in response to higher demands for bone repairing. Recently, this method, along with the emergence of functionally graded, biocompatible and biodegradable materials, has been expanded. Moreover, scaffolds with chemical, physical and external patterns have induced bone regeneration. However, the maintenance of healthy bone and its regeneration in the human body needs a series of complex and accurate processes. Hence, many studies have been accompanied for reconstructing bone by using blood-derived biomaterials, especially platelet-rich fabricates. The most important reason for using platelet-rich formulations in bone regeneration is based on releasing growth factors from alpha granules in platelets, which can induce osteogenesis. Moreover, the presence of fibrin nano-fiber structures as a constituent can provide a good substrate for cell attachments. This study attempts to review the history, structure, and biology of platelet-rich fibrin (PRF) as well as in vitro, pre-clinical, and clinical studies on the use of PRF for bone regeneration.

Fracture Resistance of Immature Incisors Following Root Filling with Various Bioactive Endodontic Cements Using an Experimental Bovine Tooth Model.

OBJECTIVE: The aim of this study was to compare the fracture resistance of immature bovine roots when using ProRoot MTA, CEM Cement, and Biodentine as root filling materials. MATERIALS AND METHODS: An immature bovine tooth model was developed by removing the coronal and apical portions of 70 bovine incisors 8 mm above and 12 mm below the cementoenamel junction (CEJ). The specimens were then divided into five groups: ProRoot MTA, CEM Cement, Biodentine, gutta-percha/AH26 sealer, and control. All groups received a 5-mm apical plug with a temporary restorative material. Then, the remaining root canal space was filled with one of the afore-mentioned materials. After setting, the specimens were mounted in acrylic resin. Then, 3 mm coronal to the CEJ from the buccal side of the teeth and at a 135°angle to the long axis, the specimens were loaded until fracture. RESULTS: The specimens in the Biodentine (2196 N) and ProRoot MTA (2103 N) groups had significantly greater fracture resistance in comparison to the control group (p = 0.01). No significant difference was found between CEM Cement, gutta-percha and sealer AH26, and control groups. No significant differences occurred between the four experimental groups (p = 0.45). CONCLUSION: Filling the root canal space with ProRoot MTA and Biodentine contributed to higher fracture resistance values.

Dental Pulp Response to RetroMTA after Partial Pulpotomy in Permanent Human Teeth.

INTRODUCTION: A lack of information exists regarding the efficacy of RetroMTA (BioMTA, Seoul, Korea) directly applied on the pulp in vital pulp therapy. This study was designed to examine the clinical efficacy of RetroMTA compared with ProRoot mineral trioxide aggregate (MTA) (Dentsply Tulsa Dental, Tulsa, OK) for partial pulpotomy. METHODS: Partial pulpotomy was performed in 22 healthy human maxillary and mandibular third molars planned for extraction. The teeth were randomly divided into 2 groups (n = 11) and underwent partial pulpotomy with RetroMTA and ProRoot MTA as the control. The teeth were then restored with glass ionomer cement. Clinical and electric pulp tests were performed after 1 and 8 weeks. The teeth were radiographed and extracted at 8 weeks. Histologic sections were prepared and analyzed for pulp inflammation and dentinal bridge formation. Data were analyzed using the Mann-Whitney U test. RESULTS: Clinical examination after 1 and 8 weeks showed no sensitivity to heat, cold, or palpation in the ProRoot MTA and RetroMTA groups. Periapical radiographs taken before the extraction of teeth showed no evidence of periapical pathology. Electric pulp testing revealed no sensitivity. Data comparisons using the Mann-Whitney U test showed no significant difference between the materials with regard to the pulp inflammation type, intensity and extension (P = .3), or bridge continuity (P = .12). However, these data revealed a significant difference between the 2 materials in pulp morphology (P < .05) and bridge thickness (P < .01). CONCLUSIONS: This is the first work to evaluate a RetroMTA histologic outcome in partial pulpotomy in human permanent teeth. It shows pulp disorganization, an absence of inflammation, and discontinuous mineralization, which may represent a potential drawback with RetroMTA in this indication.

The Micro-Shear Bond Strength of Various Resinous Restorative Materials to Aged Biodentine.

INTRODUCTION: The type of materials and application time of veneering restorations on calcium silicate cements are important factors which influence the interfacial properties. The aim of this study was to measure the micro-shear bond strength of a resin composite (RC) using several adhesive systems and a resin-modified glass ionomer cement (RM-GIC) to different aged Biodentine specimens. METHODS AND MATERIALS: A total of 15 Biodentine blocks were prepared and assigned to three aging periods: 12 min, one week and one month. Then they were subdivided into five sub-groups to receive cylinders of resinous materials. RC was applied using different adhesive systems: A) no adhesive B) etch and rinse C) two-step self-etch and D) universal adhesive in self-etch mode and E) RM-GIC applied directly over Biodentine. Micro-shear bond strength was measured and the data were analyzed using one-way and two-way ANOVA. The level of significance was set at 0.05. RESULT: There was significant interaction between Biodentine aging periods and resinous materials (P<0.05). The highest value was obtained in group D bonded to the recently set Biodentine. Increasing the aging period to one week resulted in increased micro-shear bond strength in all groups expect for group D. One-month incubation time led to reduced shear bond strength in group A, C and D. Micro-shear bond strength values of group E increased to the longer aged Biodentine. CONCLUSION: Group D showed the highest bond strength to freshly mixed Biodentine.

X-ray diffraction analysis of MTA mixed and placed with various techniques.

OBJECTIVES: The aim of this study was to evaluate the effect of various mixing techniques as well as the effect of ultrasonic placement on hydration of mineral trioxide aggregate (MTA) using X-ray diffraction (XRD) analysis. MATERIALS AND METHODS: One gram of ProRoot MTA and MTA Angelus powder was mixed with a 0.34-g of distilled water. Specimens were mixed either by mechanical mixing of capsules for 30 s at 4500 rpm or by manual mixing followed by application of a compaction pressure of 3.22 MPa for 1 min. The mixtures were transferred into the XRD sample holder with minimum pressure. Indirect ultrasonic activation was applied to half of the specimens. All specimens were incubated at 37 °C and 100% humidity for 4 days. Samples were analyzed by XRD. Phase identification was accomplished by use of search-match software utilizing International Centre for Diffraction Data (ICDD). RESULTS: All specimens comprised tricalcium silicate, calcium carbonate, and bismuth oxide. A calcium hydroxide phase was formed in all ProRoot specimens whereas among MTA Angelus groups, it was found only in the sample mixed mechanically and placed by ultrasonication. CONCLUSIONS: Mechanical mixing followed by ultrasonication did not confer a significant disadvantage in terms of hydration characteristics of MTA. CLINICAL RELEVANCE: Clinicians vary in the way they mix and place MTA. These variations might affect their physical characteristics and clinical performance. For ProRoot MTA, the mixing and placement methods did not affect its rheological properties, whereas for MTA Angelus, mechanical mixing combined with ultrasonic placement enhanced the calcium hydroxide phase formation.

Human Pulp Responses to Partial Pulpotomy Treatment with TheraCal as Compared with Biodentine and ProRoot MTA: A Clinical Trial.

INTRODUCTION: Questions exist regarding the efficacy of resin-containing materials such as TheraCal directly applied on the pulp. This study sought to investigate the clinical efficacy of TheraCal as compared with Biodentine and ProRoot mineral trioxide aggregate (MTA) for partial pulpotomy. METHODS: In this clinical trial, partial pulpotomy was performed for 27 sound human maxillary and mandibular third molars scheduled for extraction. The teeth were randomly divided into 3 groups (n = 9) and underwent partial pulpotomy with TheraCal, Biodentine, and ProRoot MTA. The teeth were then restored with glass ionomer cement. Clinical and electric pulp tests were performed after 1 and 8 weeks. The teeth were radiographed and extracted at 8 weeks. Histologic sections were prepared and analyzed for pulp inflammation and dentinal bridge formation. Data were analyzed by using one-way analysis of variance. RESULTS: Clinical examination showed no sensitivity to heat, cold, or palpation in ProRoot MTA and Biodentine groups. Two patients in TheraCal group (20%) reported significant pain at 1 week. Periapical radiographs showed no periapical pathology, and electric pulp test revealed a normal pulp response with no hypersensitivity. Inflammation was absent with all materials at 8 weeks. Normal pulp organization was seen in 33.33% of the teeth in ProRoot MTA, 11.11% in TheraCal, and 66.67% in Biodentine group (P = .06). Biodentine group showed complete dentinal bridge formation in all teeth, whereas this rate was 11% and 56% in TheraCal and ProRoot MTA groups, respectively (P = .001). CONCLUSIONS: Overall, Biodentine and MTA performed better than TheraCal when used as partial pulpotomy agent and presented the best clinical outcomes.

Second-generation Platelet Concentrate (Platelet-rich Fibrin) as a Scaffold in Regenerative Endodontics: A Case Series.

INTRODUCTION: The purpose of this case series was to report the clinical and radiographic results of a pulp regenerative procedure using platelet-rich fibrin (PRF), a second-generation platelet concentrate, in immature teeth with necrotic pulps. METHODS: Root canal revascularization using PRF was performed on 4 immature teeth with necrotic pulps. After access cavity preparation, the root canals were irrigated with low concentration sodium hypochlorite solution (1.5% sodium hypochlorite [20 mL/canal, 5 minutes]) and then irrigated with saline (20 mL/canal, 5 minutes). Equal proportions (167 mg) of ciprofloxacin, metronidazole, and cefaclor were mixed and diluted to a final concentration of 1 g/mL. Finally, the canal was sealed with 3-4 mm of a temporary restorative material, and patients were dismissed for 2 to 3 weeks. At the second appointment, 9 mL of the patient's whole blood was obtained and centrifuged to prepare a PRF clot. Canals were irrigated with 17% EDTA, and a sharp spreader was inserted beyond the apex. Then, the PRF clot was placed inside the root canals, and Biodentine (Septodont, Saint-Maur, France) was placed directly over the PRF. The teeth were restored permanently with glass ionomer cement and composite resin. RESULTS: Clinical examinations revealed that all cases were asymptomatic at the recall appointments at 1, 3, 6, 12, and 18 months. Radiographs revealed resolution of the periapical lesions, further root development, and apical closure in all cases. CONCLUSIONS: On the basis of the short-term results up to 12 months, PRF clots acted as successful scaffolds for the regeneration of pulpal contents in immature teeth with necrotic pulps.

Effect of varying water-to-powder ratios and ultrasonic placement on the compressive strength of mineral trioxide aggregate.

INTRODUCTION: The purpose of this study was to compare the compressive strength of mineral trioxide aggregate (MTA) when mixed with 2 different water-to-powder (WP) proportions using either hand or ultrasonic placement. METHODS: Tooth-colored ProRoot MTA (Dentsply Maillefer, Ballaigues, Switzerland) and white MTA Angelus (Angelus Soluçoes Odontologicas, Londrina, Brazil) were investigated. One gram of each MTA powder was mixed with either 0.34 or 0.40 g distilled water. The 4 groups were further divided into 2 groups of 5 specimens for each of the following techniques: conventional (ie, hand placement) and placement using indirect ultrasonic activation for 30 seconds. All specimens were subjected to compressive strength testing after 4 days. The results were statistically analyzed with multivariate analysis of variance and Tukey Honestly Significant Difference tests at a significance level of P < .05. RESULTS: The mean compressive strength values of ProRoot MTA (84.17 ± 22.68) were significantly greater than those of MTA Angelus (47.71 ± 14.29) (P < .01). Specimens mixed with the 0.34 WP ratio had higher compressive strength values (72.85 ± 25.77) than those mixed with the 0.40 WP ratio (56.69 ± 24.85) (P < .05). The highest compressive strength values were recorded for ProRoot MTA specimens that were mixed in the 0.34 WP ratio, and then the samples were placed with ultrasonic activation (mean = 91.35 MPa). The lowest values were recorded for MTA Angelus samples that were mixed in the 0.40 WP ratio, and the specimens were placed without ultrasonic activation (mean = 36.36 MPa). Ultrasonic activation had no significant difference in terms of compressive strength. CONCLUSIONS: When using ProRoot MTA and MTA Angelus, higher WP ratios resulted in lower compressive strength values. Ultrasonication had no significant effect on the compressive strength of the material regardless of the WP ratio that was used. Therefore, adherence to the manufacturer's recommended WP ratio when preparing MTA for use in dental applications is advised.

Influence of tooth length on the accuracy of the Root ZX electronic apical foramen locator: an ex vivo study.

OBJECTIVE: Electronic apical foramen locators are now widely used to determine working length. This study was designed to determine whether tooth length influenced the accuracy of the Root ZX device. MATERIALS AND METHODS: Forty extracted maxillary canine teeth with a length range of 27-29 mm were selected. Access cavities were prepared and coronal flaring of canals performed. The teeth were mounted in self-polymerizing acrylic resin to facilitate horizontal sectioning except for the apical 3-4-mm portion of the root and embedded in alginate as the electronic medium. Electronic measurements were taken at the major foramen, 'zero' reading using the Root ZX and compared with the actual root canal length. The teeth were sectioned 3 mm from the coronal reference point to create a second group with shorter length; these reductions in the length continued six times in all to create seven groups of 40 specimens each. The actual and electronic lengths of specimens in each group were measured. Data were analyzed by Pearson's correlation coefficient. RESULTS: Identical measurements between the actual and electronic root canal length from the longest to the shortest groups were 12.5%, 10.0%, 20.0%, 27.5%, 37.5%, 35.0% and 45.0%, respectively. There was a mild negative correlation between the precise measurements of the Root ZX and root canal lengths in the seven groups (r = -0.964, p < 0.001). CONCLUSION: Under the conditions of the study, the Root ZX device was more accurate in shorter teeth compared to longer ones.

Surface microhardness of three thicknesses of mineral trioxide aggregate in different setting conditions.

OBJECTIVES: This study aimed to compare the surface microhardness of mineral trioxide aggregate (MTA) samples having different thicknesses and exposed to human blood from one side and with or without a moist cotton pellet on the other side. MATERIALS AND METHODS: Ninety cylindrical molds with three heights of 2, 4, and 6 mm were fabricated. In group 1 (dry condition), molds with heights of 2, 4, and 6 mm (10 molds of each) were filled with ProRoot MTA (Dentsply Tulsa Dental), and the upper surface of the material was not exposed to any additional moisture. In groups 2 and 3, a distilled water- or phosphate-buffered saline (PBS)-moistened cotton pellet was placed on the upper side of MTA, respectively. The lower side of the molds in all the groups was in contact with human blood-wetted foams. After 4 day, the Vickers microhardness of the upper surface of MTA was measured. RESULTS: In the dry condition, the 4 and 6 mm-thick MTA samples showed significantly lower microhardness than the 2 mm-thick samples (p = 0.003 and p = 0.001, respectively). However, when a distilled water- or PBS-moistened cotton pellet was placed over the MTA, no significant difference was found between the surface microhardness of samples having the abovementioned three thicknesses of the material (p = 0.210 and p = 0.112, respectively). CONCLUSIONS: It could be concluded that a moist cotton pellet must be placed over the 4 to 6 mm-thick MTA for better hydration of the material. However, this might not be necessary when 2 mm-thick MTA is used.

In vitro cytotoxicity of four calcium silicate-based endodontic cements on human monocytes, a colorimetric MTT assay.

OBJECTIVES: This study was performed to evaluate the cytotoxicity of four calcium silicate-based endodontic cements at different storage times after mixing. MATERIALS AND METHODS: Capillary tubes were filled with Biodentine (Septodont), Calcium Enriched Mixture (CEM cement, BioniqueDent), Tech Biosealer Endo (Tech Biosealer) and ProRoot MTA (Dentsply Tulsa Dental). Empty tubes and tubes containing Dycal were used as negative and positive control groups respectively. Filled capillary tubes were kept in 0.2 mL microtubes and incubated at 37℃. Each material was divided into 3 groups for testing at intervals of 24 hr, 7 day and 28 day after mixing. Human monocytes were isolated from peripheral blood mononuclear cells and cocultered with 24 hr, 7 day and 28 day samples of different materials for 24 and 48 hr. Cell viability was evaluated using an MTT assay. RESULTS: In all groups, the viability of monocytes significantly improved with increasing storage time regardless of the incubation time (p < 0.001). After 24 hr of incubation, there was no significant difference between the materials regarding monocyte viability. However, at 48 hr of incubation, ProRoot MTA and Biodentine were less cytotoxic than CEM cement and Biosealer (p < 0.01). CONCLUSIONS: Biodentine and ProRoot MTA had similar biocompatibility. Mixing ProRoot MTA with PBS in place of distilled water had no effect on its biocompatibility. Biosealer and CEM cement after 48 hr of incubation were significantly more cytotoxic to on monocyte cells compared to ProRoot MTA and Biodentine.

Effect of acidic environment on dislocation resistance of endosequence root repair material and mineral trioxide aggregate.

OBJECTIVE: The aim of this study was to compare the effect of an acidic environment on dislocation resistance (push-out bond strength) of EndoSequence Root Repair Material (ERRM putty and ERRM paste), a new bioceramic-based material, to that of mineral tri-oxide aggregate (MTA). MATERIALS AND METHODS: One-hundred twenty root dentin slices with standardized canal spaces were divided into 6 groups (n = 20 each) and filled with tooth-colored ProRoot MTA (groups 1 and 2), ERRM putty (groups 3 and 4), or ERRM paste (groups 5 and 6). The specimens of groups 1, 3, and 5 were exposed to phosphate buffered saline (PBS) solution (pH=7.4) and those of groups 2, 4, and 6 were exposed to butyric acid (pH= 4.4). The specimens were then incubated for 4 days at 37°C. The push-out bond strength was then measured using a universal testing machine. Failure modes after the push-out test were examined under a light microscope at ×40 magnification. The data for dislocation resistance were analyzed using the t-test and one-way analysis of variance. RESULTS: In PBS environment (pH=7.4), there were no significant differences among materials (P=0.30); but the mean push-out bond strength of ERRM putty was significantly higher than that of other materials in an acidic environment (P<0.001). Push-out bond strength of MTA and ERRM paste decreased after exposure to an acidic environment; whereas ERRM putty was not affected by acidic pH. The bond failure mode was predominantly cohesive for all groups except for MTA in an acidic environment; which showed mixed bond failure in most of the specimens. CONCLUSION: The force needed for dislocation of MTA and ERRM paste was significantly lower in samples stored in acidic pH; however, push-out bond strength of ERRM putty was not influenced by acidity.