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Objectives: This study aimed to investigate the influence of access cavity designs on the mechanical properties of a single-rooted mandibular first premolar tooth under various static loads using a finite element analysis., Materials and Methods: 3-dimensional FEA designs were modeled according to the access cavity designs: an intact tooth (control), traditional access cavity (TEC-I), traditional access cavity with Class-II mesio-occlusal cavity design (TEC-II), conservative access cavity (CEC), ninja access cavity (NEC), caries-driven access cavity (Cd-EC), buccal access cavity (BEC) and bucco-occlusal access cavity (BOEC). After the simulated access cavity preparations, root canal treatment was simulated and three different static loads which mimicked oblique and vertical mastication forces were applied to the models. The stress distribution and maximum Von Misses stress values were recorded. The maximum stress values were obtained on both enamel and dentin under multi-point vertical loads., Results: The maximum stress values were obtained on both enamel and dentin under multi-point vertical loads. Under all load types, the minimum stress distribution was observed in the control group, followed by CEC, NEC and BEC designs. The highest stress concentration was detected in Cd-EC and TEC-II designs. Under single-point vertical loading, the stress was mostly concentrated in the lingual PCD area, while under multi-point vertical loading, the entire root surface was stress-loaded except for the lingual apical third of the root., Conclusion: Preserving tooth tissue by simulating CEC, NEC and BEC access cavities increased the load capacity of a single-rooted mandibular first premolar following simulated endodontic treatment., (© 2024. The Author(s).)

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., (© 2023. The Author(s).)

Aim: The study aimed to develop a bicomponent bioactive hydrogel formed in situ and enriched with an extract of platelet-rich fibrin (PRFe) and to assess its potential for use in pulp-dentine complex tissue engineering via cell homing., Methodology: A bicomponent hydrogel based on photo-activated naturally derived polymers, methacrylated chitosan (ChitMA) and methacrylated collagen (ColMA), plus PRFe was fabricated. The optimized formulation of PRFe-loaded bicomponent hydrogel was determined by analysing the mechanical strength, swelling ratio and cell viability simultaneously. The physical, mechanical, rheological and morphological properties of the optimal hydrogel with and without PRFe were determined. Additionally, MTT, phalloidin/DAPI and live/dead assays were carried out to compare the viability, cytoskeletal morphology and migration ability of stem cells from the apical papilla (SCAP) within the developed hydrogels with and without PRFe, respectively. To further investigate the effect of PRFe on the differentiation of encapsulated SCAP, alizarin red S staining, RT-PCR analysis and immunohistochemical detection were performed. Statistical significance was established at p <  .05., Results: The optimized formulation of PRFe-loaded bicomponent hydrogel can be rapidly photocrosslinked using available dental light curing units. Compared to bicomponent hydrogels without PRFe, the PRFe-loaded hydrogel exhibited greater viscoelasticity and higher cytocompatibility to SCAP. Moreover, it promoted cell proliferation and migration in vitro. It also supported the odontogenic differentiation of SCAP as evidenced by its promotion of biomineralization and upregulating the gene expression for ALP, COL I, DSPP and DMP1 as well as facilitated angiogenesis by enhancing VEGFA gene expression., Conclusions: The new PRFe-loaded ChitMA/ColMA hydrogel developed within this study fulfils the criteria of injectability, cytocompatibility, chemoattractivity and bioactivity to promote odontogenic differentiation, which are fundamental requirements for scaffolds used in pulp-dentine complex regeneration via cell-homing approaches., (© 2022 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objectives: To investigate the effects of blood and platelet-rich fibrin (PRF), commonly used scaffolds in regenerative endodontic treatment (RET), on the hydration, microstructure, and color stability of three hydraulic calcium silicate cements (HCSCs), OrthoMTA, RetroMTA, and TotalFill-BC-RRM., Materials and Methods: The HCSCs were prepared and placed into polyethylene molds and transferred to Eppendorf tubes containing PRF, blood, or PBS and then incubated for 1 week or 1 month. The microstructure and hydration of the cements were studied by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD). The chromatic alteration of materials was also measured using a spectrophotometer. The data for color stability were analyzed using 2-way analysis of variance and Tukey post hoc tests., Results: There was no significant difference between the color stability of cements exposed to PBS (p > 0.05). The chromatic alteration of cements exposed to blood was significantly greater than those exposed to PRF and PBS (p < 0.001). In the presence of blood and PRF, the color change of OrthoMTA was significantly greater than that of RetroMTA and TotalFill (p < 0.05), with no significant difference between RetroMTA and TotalFill (p > 0.05). XRD analysis of all cements revealed a calcium hydroxide peak after 1-week and 1-month exposure to the media; however, OrthoMTA and TotalFill exposed to blood and PRF for 1 month showed weaker calcium hydroxide peaks. SEM images revealed cements exposed to PBS had a different surface microstructure compared to those exposed to blood and PRF. Furthermore, the surface microstructure of HCSCs was influenced by the type of cement radiopacifier (bismuth oxide or zirconium oxide). EDS analysis of the elemental composition in all groups displayed peaks of Ca, O, C, Si, P, and Al., Conclusions: Color stability, hydration behavior, and microstructure of HCSCs were affected by exposure to PRF and blood and the type of cement radiopacifier., Clinical Relevance: As some important physicochemical properties of HCSCs could be influenced by the environmental conditions and the type of radiopacifier, alternatives to blood clot and HCSCs containing substitutes for bismuth oxide might be more suitable in RETs., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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., Competing Interests: Conflict of interestThe authors have no relevant financial or non-financial interests to disclose., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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., Competing Interests: The authors declare no conflict of interest., (© 2023 The Author(s).)

Background: Pulp-dentine complex regeneration via tissue engineering is a developing treatment modality that aims to replace necrotic pulps with newly formed healthy tissue inside the root canal. Designing and fabricating an appropriate scaffold is a crucial step in such a treatment., Objectives: The present study aimed to review recent advances in the design and fabrication of scaffolds for de novo regeneration of pulp-dentine complexes via tissue engineering approaches., Methods: A literature search was conducted using PubMed, Europe PMC, Scopus and Google Scholar databases. To highlight bioengineering techniques for de novo regeneration of pulp-dentine complexes, both in vitro and in vivo studies were included, and clinical studies were excluded., Results: In the present review, four main classes of scaffolds used to engineer pulp-dentine complexes, including bioceramic-based scaffolds, synthetic polymer-based scaffolds, natural polymer-based scaffolds and composite scaffolds, are covered. Additionally, recent advances in the design, fabrication and application of such scaffolds are analysed along with their advantages and limitations. Finally, the importance of vascular network establishment in the success of pulp-dentine complex regeneration and strategies used to create scaffolds to address this challenge are discussed., Discussion: In the tissue engineering platform, scaffolds provide structural support for cells to adhere and proliferate and also regulate cell differentiation and metabolism. Up to now, considerable progress has been achieved in the field of pulp-dentine complex tissue engineering, and a spectrum of scaffolds ranging from bioceramic-based to naturally derived scaffolds has been fabricated. However, in designing a suitable scaffold for engineering pulp-dentine complexes, a variety of characteristic parameters related to biological, structural, physical and chemical features should be considered., Conclusion: The variety of biomaterials and fabrication techniques provides a great opportunity to address some of the requirements for scaffolds in regenerative endodontics. However, more studies are required to develop an ideal scaffold for use in a clinical setting., (© 2022 British Endodontic Society. Published by John Wiley & Sons Ltd.)

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 .

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 ., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Conservative Dentistry.)

The treatment of deep carious lesions involves the use of ion-releasing agents to seal the lesions. These agents release minerals, leading to the remineralization of the remaining demineralized dentin. This study aimed to compare the dentin caries remineralization with bioactive cements. 60 Dentin blocks were prepared from the dentin of human third molars. Artificial carious lesions were induced on the blocks with pH cycling. The samples were divided into five groups (n=12). Dycal, Oxford ActiveCal PC, Biodentine, and ACTIVA BioACTIVE were applied using a mold. One group did not receive any cement. The samples were stored in remineralization solution for 30 days. The cement was removed using a #15 blade, and the dentin surface was evaluated using Energydispersive X-ray Spectroscopy and X-ray Diffraction. One-way ANOVA did not show a significant difference in the weight percentages of calcium and phosphorus and the calcium-to-phosphorus ratios between the groups. The highest and the lowest weight percentages of calcium and phosphorus were observed in Biodentine and control groups, respectively. There were no significant differences in the remineralization properties of bioactive cements. Hydroxyapatite crystals were not formed in any of the adjacent dentin using these cements., (Copyright© 2022 Dennis Barber Ltd.)

Background: One problem encountered in vital pulp therapy is the dislodgment of biomaterial as the result of the application of mechanical condensation forces for the final restoration of the cavity or occlusal loads., Material and Methods: In this in vitro , experimental study, 90 dentin discs were prepared with Gates Glidden drills to have a 1.3 mm canal diameter. The specimens were divided into nine groups (n=10). ProRoot MTA, Biodentine, and TheraCal were applied in groups 1-3, 4-6, and 7-9, respectively. The PBS was measured after 15 minutes, four hours, and three days and mode of failure was determined., Results: The interaction effect of time and material on PBS was statistically significant ( P <0.003). The PBS of Biodentine and ProRoot MTA significantly increased over time ( P =0.000). At 15 minutes and four hours, the PBS of TheraCal was higher than that of Biodentine and ProRoot MTA ( P =0.000). Our results showed the predominant type of bond failures in Biodentine and Theracal groups was cohesive, whereas it was adhesive for ProRootMTA., Conclusions: Theracal showed higher values of bond strength than Biodentine and ProRootMTA at 15 minutes & four hours and may thus be better options for single session of VPT. Key words: Biodentine, Mineral Trioxide Aggregate, Pushout Bond Strength, TheraCal., Competing Interests: Conflicts of interest No potential conflict of interest relevant to this article was reported., (Copyright: © 2021 Medicina Oral S.L.)

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., (Copyright © 2021 Elsevier B.V. All rights reserved.)