Knowledge of undergraduate and postgraduate dental students about managing traumatic dental injuries based on the 2020 International Association of Dental Traumatology guidelines: A online survey of 10 dental schools from 10 countries.

BACKGROUND/AIM: The knowledge of dental students about managing traumatic dental injuries (TDIs) may not be uniform, depending on global location and dental education. The aim of this study was to evaluate the level of knowledge of undergraduate and postgraduate students specializing in endodontics and pediatric dentistry at 10 dental schools in 10 countries about the 2020 International Association of Dental Traumatology (IADT) guidelines regarding the management of TDIs. MATERIALS & METHODS: A previously published questionnaire was used in the current survey. It was an online survey with 12 questions regarding the management of TDIs and some additional questions regarding sociodemographic and professional profiles of the participants were added. The survey was distributed to final-year undergraduate students and postgraduate students in pediatric dentistry and endodontics from 10 dental schools. Simple frequency distributions and descriptive statistics were predominantly used to describe the data. Differences in the median percentage scores among the student categories were assessed using the Kruskal-Wallis test followed by Dwass-Steel-Critchlow-Fligner pairwise comparisons. RESULTS: A total of 347 undergraduates, 126 postgraduates in endodontics, and 72 postgraduates in pediatric dentistry from 10 dental schools participated in this survey. The postgraduates had a significantly higher percentage score for correct responses compared with the undergraduates. No significant difference was observed between the endodontic and pediatric dentistry postgraduates. CONCLUSION: The knowledge possessed by undergraduate and postgraduate students concerning the IADT-recommended management of TDIs varied across the globe and some aspects were found to be deficient. This study emphasizes the critical importance of reassessing the teaching and learning activities pertaining to the management of TDIs.

Characterization of nano-hydroxyapatite incorporated carboxymethyl chitosan composite on human dental pulp stem cells.

AIM: To compare the odontogenic differentiation potential of a composite scaffold (CSHA) comprising of nano-hydroxyapatite (nHAp) and carboxymethyl chitosan (CMC) with Biodentine on human dental pulp stem cells (hDPSCs). METHODOLOGY: A CSHA scaffold was prepared through an ultrasonication route by adding nHAp and CMC (1:5 w/w) in water medium followed by freeze-drying. Physicochemical characterization was achieved using scanning electron microscopy with energy-dispersive X-ray spectroscopy, X-ray diffraction and Fourier transform infrared spectroscopy. In-vitro bioactivity and pH assessments were done by soaking in simulated body fluid (SBF) for 28 days. The angiogenic and odontogenic differentiation abilities were assessed by expression of vascular endothelial growth factor (VEGF) and Dentine sialophosphoprotein (DSPP) markers on cultured hDPSCs by flow cytometry and RT-qPCR at 7, 14 and 21 days. Cell viability/proliferation and biomineralization abilities of CSHA were compared with Biodentine by MTT assay, alkaline phosphatase (ALP) activity, Alizarin Red Staining (ARS) and osteopontin (OPN) expression on hDPSCs following 7 and 14 days. Data were statistically analysed with Kruskal Wallis and Friedman tests as well as one way anova followed by appropriate post hoc tests (p < .05). RESULTS: Characterization experiments revealed a porous microstructure of CSHA with pore diameter ranging between 60 and 200 µm and 1.67 Ca/P molar ratio along with the characteristic functional groups of both HAp and CMC. CSHA displayed bioactivity in SBF by forming apatite-like crystals and maintained a consistent pH value of 7.70 during 28 days' in vitro studies. CSHA significantly upregulated VEGF and DSPP levels on hDPSCs on day 21 compared with day 7 (p < .05). Further, CSHA supported cell viability/proliferation over 14 days like Biodentine with no statistical differences (p > .05). However, CSHA exhibited increased ALP and ARS activity with an intense OPN staining compared with Biodentine after 14 days (p < .05). CONCLUSION: The results highlighted the odontogenic differentiation and biomineralization abilities of CSHA on hDPSCs with significant VEGF and DSPP gene upregulations. Further, CSHA exhibited enhanced mineralization activity than Biodentine, as evidenced by increased ALP, ARS and OPN activity on day 14. The nHAp-CMC scaffold has the potential to act as an effective pulp capping agent; however, this needs to be further validated through in-vivo animal studies.

Eggshell derived nano-hydroxyapatite incorporated carboxymethyl chitosan scaffold for dentine regeneration: A laboratory investigation.

AIM: To assess odontogenic differentiation abilities of porous biomineralizable composite scaffolds comprising eggshell derived nano-hydroxyapatite (HAnp) and carboxymethyl chitosan (CMC) on cultured human dental pulp stem cells (hDPSCs). METHODOLOGY: Nano-hydroxyapatite was derived from eggshells using a simple combustion method and CMC was prepared from chitosan through a chemical route. Several compositions of HAnp-CMC (0:5, 5:0, 1:5, 2:5, 3:5, 4:5 and 1:1 w/w%) scaffolds were prepared by magnetic stirring and freeze-drying methods. HAnp-CMC scaffolds were characterized using high-resolution scanning electron microscopy combined with energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy and X-ray diffraction methods. In vitro bioactivity was determined following the interaction in simulated body fluid for 21 days. The optimized composite was then loaded onto hDPSCs to assess cell viability/proliferation, dentine sialophosphoprotein (DSPP) and vascular endothelial growth factor (VEGF) expressions using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, real-time quantitative polymerase chain reaction and flow cytometry methods, respectively, following 7, 14 and 21 days. For intergroup and intragroup comparisons, Kruskal-Wallis and Friedman tests were employed, respectively, followed by appropriate post hoc test (Dunn). Significant levels were set at *p < .05 and *p < .01. RESULTS: Synthesized hydroxyapatite (HAp) comprised crystals ranging from 20 to 50 nm (HAnp) with spherulite morphology and calcium/phosphorus (Ca/P) molar ratio of 1.67. The ultrastructure of all the scaffolds revealed a highly interconnected porous microstructure, whilst the chemical characterization displayed specific functional groups of both HAnp and CMC. In vitro bioactivity assessment confirmed the biomineralization potential of all scaffolds with an apatite-like crystal formation on the surface. The 1:5 HAnp-CMC revealed a favourable pore size (60-180 µm) that was suitable for cell seeding and was chosen for further experiments. Cell viability/proliferation rates of hDPSCs loaded 1:5 HAnp-CMC at 21st day was significantly greater than that at 7th day (p < .05). The mean relative quantification of DSPP expression by the scaffold was significantly higher (p < .05) on day 21 (3.16) than on day 7 (1.67). Mean fluorescence intensity of the VEGF expression at day 21 (32.5) was also significantly higher (p < .01) than at day 7 (12.54). CONCLUSION: hDPSCs on 1:5 HAnp-CMC scaffolds displayed increased cell viability/proliferation and enhanced DSPP as well as VEGF expressions. The 1:5 HAnp-CMC composite has the potential to serve as a promising scaffold for dentine regeneration.

Human amniotic membrane extracellular matrix scaffold for dental pulp regeneration in vitro and in vivo.

AIM: In order to obtain a 3-dimentional scaffold with predictable clinical results for pulp regeneration, this study aims to fabricate and characterize a porous decellularized human amniotic membrane (HAM) extracellular matrix (ECM) scaffold, and evaluate its potential to promote pulp regeneration in vitro and in vivo. METHODOLOGY: The HAM was decellularized, and its histology and DNA content were analysed to confirm decellularization. The scaffolds were synthesized with 15, 22.5 and 30 mg/ml concentrations. The porosity, pore size, phosphate-buffered saline (PBS) absorption and degradation rate of the scaffolds were assessed. In vitro experiments were performed on human dental pulp stem cells (hDPSCs) to assess their viability, proliferation, adhesion and migration on the scaffolds. The optimal group was selected for in vivo immunogenicity assessment and was also used as the cell-free or cell-loaded scaffold in root segment models to evaluate pulp regeneration. All nonparametric data were analysed with the Kruskal-Wallis test followed by Dunn's post hoc test, whilst quantitative data were analysed with one-way anova. RESULTS: Decellularization of HAM was confirmed (p < .05). The porosity of all scaffolds was more than 95%, and the pore size decreased with an increase in ECM concentration (p < .01). PBS absorption was not significantly different amongst the groups, whilst 30 mg/ml ECM scaffold had the highest degradation rate (p < .01). The hDPSCs adhered to the scaffold, whilst their proliferation rate increased over time in all groups (p < .001). Cell migration was higher in 30 mg/ml ECM scaffold (p < .05). In vivo investigation with 30 mg/ml ECM scaffold revealed mild to moderate inflammatory response. In root segments, both cell-free and cell-loaded 30 mg/ml scaffolds were replaced with newly formed, pulp-like tissue with no significant difference between groups. Immunohistochemical assessments revealed high revascularization and collagen content with no significant difference amongst the groups. CONCLUSION: The 30 mg/ml HAM ECM scaffold had optimal physical properties and better supported hDPSC migration. The HAM ECM scaffold did not interfere with formation of pulp-like tissue and revascularization within the root canal when employed as both cell-free and cell-loaded scaffold. These results highlight the potential of HAM ECM membrane for further investigations in regenerative endodontics.

Methodological quality assessment criteria for the evaluation of laboratory-based studies included in systematic reviews within the specialty of Endodontology: A development protocol.

High-quality systematic reviews in the field of Dentistry provide the most definitive overarching evidence for clinicians, guideline developers and healthcare policy makers to judge the foreseeable risks, anticipated benefits, and potential harms of dental treatment. In the process of carrying out a systematic review, it is essential that authors appraise the methodological quality of the primary studies they include, because studies which follow poor methodology will have a potentially serious negative impact on the overall strength of the evidence and the recommendations that can be drawn. In Endodontology, systematic reviews of laboratory studies have used quality assessment criteria developed subjectively by the individual authors as there are no comprehensive, well-structured, and universally accepted criteria that can be applied objectively and universally to individual studies included in reviews. Unfortunately, these subjective criteria are likely to be inaccurately defined, unreliably applied, inadequately analysed, unreasonably biased, defective, and non-repeatable. The aim of the present paper is to outline the process to be followed in the development of comprehensive methodological quality assessment criteria to be used when evaluating laboratory studies, that is research not conducted in vivo on humans or animals, included in systematic reviews within Endodontology. The development of new methodological quality assessment criteria for appraising the laboratory-based studies included in systematic reviews within Endodontology will follow a three-stage process. First, a steering committee will be formed by the project leaders to develop a preliminary list of assessment criteria by modifying and adapting those already available, but with the addition of several new items relevant for Endodontology. The initial draft assessment criteria will be reviewed and refined by a Delphi Group (n = 40) for their relevance and inclusion using a nine-point Likert scale. Second, the agreed items will then be discussed in an online or face-to-face meeting by a group of experts (n = 10) to further refine the assessment criteria. Third, based on the feedback received from the online/face-to-face meeting, the steering committee will revise the quality assessment criteria and subsequently a group of authors will be selected to pilot the new system. Based on the feedback collected, the criteria may be revised further before being approved by the steering committee. The assessment criteria will be published in relevant journals, presented at national and international congresses/meetings, and will be freely available on a dedicated website. The steering committee will update the assessment criteria periodically based on feedback received from end-users.

Biofilm formation following chitosan-based varnish or chlorhexidine-fluoride varnish application in patients undergoing fixed orthodontic treatment: a double blinded randomised controlled trial.

BACKGROUND: Orthodontic treatment poses an increased risk of plaque accumulation and demineralisation of enamel leading to white spot lesion around the brackets. This parallel arm trial aims to assess the degree of bacterial plaque formation adjacent to orthodontic brackets, following the application of a chitosan-based varnish or chlorhexidene-fluoride varnish. METHODS: A total of 200 teeth from 20 patients undergoing fixed orthodontic therapy were assessed and biofilm formation around the brackets were recorded using the Bonded Bracket Index (Plaque index) at baseline and weekly for 6 weeks. The bacterial count and plaque pH at corresponding weekly intervals were also recorded. Following bracket bonding, the patients were cluster randomised to receive chitosan-based varnish-CHS (UNO Gel Bioschell, Germiphene corp., Brantford, Canada) or chlorhexidine-fluoride varnish-CFV (Cervitec F, Ivoclar Vivadent, Schaan, Liechtenstein) every week on the representative teeth respectively. BBI proportions were compared between groups at all time intervals using Chi square test. Mean plaque bacterial count and plaque pH were compared using Mann Whitney U test and Tukey's HSD test respectively. RESULTS: Baseline characteristics were similar between the groups: Mean age was CHS = 23 and CFV = 21; male to female ratio was CHS = 5/5, CFV = 7/3. At the end of 6 weeks, chitosan-based varnish performed equal to chlorhexidine-fluoride varnish (P > 0.05) with 98% and 95% of teeth with acceptable scores respectively. The plaque bacterial count significantly reduced at 6 weeks for both varnish compared to the baseline; The value for CHS was 0.43 ± 0.4 × 104 and CFV was 0.77 ± 0.64 × 104 CFU (P < 0.05), with no difference between both the varnishes. Both varnishes had no effect on the plaque pH that remained neutral. CONCLUSION: This trial showed that both chitosan-based varnish and chlorhexidine-fluoride varnish reduced bacterial count, while the plaque pH remained neutral over a period of six weeks in patients undergoing fixed orthodontic therapy. The anti-plaque effects of the natural biopolymeric chitosan-based varnish was similar to that of chlorhexidine-fluoride varnish, a known chemotherapeutic agent. Registration: This trial protocol was registered with https://www.ctri.nic.in (CTRI/2019/05/018896). (Date of registration 02/05/2019). PROTOCOL: The protocol was not published before trial commencement.

A chitosan-based irrigant improves the dislocation resistance of a mineral trioxide aggregate-resin hybrid root canal sealer.

OBJECTIVES: To investigate the effectiveness of root canal irrigation with chitosan on the dislocation resistance of a root canal sealer (MTA Fillapex) in vitro, measured by the push-out bond strength test. MATERIALS AND METHODS: Root canals of mandibular premolars (n = 57) were prepared using rotary files with 5.25% sodium hypochlorite as the irrigant during instrumentation. Following this, the specimens were randomly divided into three groups (n = 19) based on the final irrigant: group 1, 0.2% chitosan solution; group 2, 17% EDTA solution; group 3, saline. Three specimens from each group were analyzed using scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS). The remaining specimens of each group were divided into two subgroups (n = 8) based on the method of agitation of the final irrigants (chitosan/EDTA/saline): subgroup A, sonic (Endoactivator, Dentsply Maillefer); subgroup B, no activation (control). After irrigation, all specimens obturated with a commercial mineral trioxide aggregate-resin hybrid sealer (MTA Fillapex, Angelus, Londrina, Brazil). Dislocation resistance was measured using the push-out bond strength test after 3 weeks. The data were analyzed using Kruskal-Wallis test (P = 0.05). RESULTS: Immaterial of the irrigant agitation, groups irrigated with chitosan showed significantly higher bond strength values than those irrigated with EDTA (P < 0.05). Groups irrigated with saline showed the least bond strength values (P < 0.05). When EDTA was used, sonic agitation significantly improved the bond strength of the sealer, compared to the control (P < 0.05). There was no significant difference between sonic agitation and the control when chitosan solution was used as the final irrigant (P > 0.05). The nitrogen/carbon ratio was significantly higher in chitosan groups compared to the control group (P < 0.05). CONCLUSION: This study provides the first evidence that chitosan irrigation improves the dislocation resistance of MTA-resin hybrid root canal sealer, compared to EDTA and saline irrigation. CLINICAL RELEVANCE: Chitosan-based irrigation has been previously shown to demonstrate anti-biofilm properties in the root canal. The present study demonstrates that chitosan can improve the bond strength of a root filling material, which may contribute to better sealing of the root canal system.

Temporal-controlled bioactive molecules releasing core-shell nano-system for tissue engineering strategies in endodontics.

Temporal-controlled release of bioactive molecules is of key importance in the clinical translation of tissue engineering techniques. We engineered a core-shell nano-system (TD-NS) that sequentially released transforming growth factor-ß1 (TGF-ß1), a chemotactic/proliferating growth factor and dexamethasone (Dex), an osteo/odontogenic agent in a temporal-controlled manner. In stage-1, there was a rapid release of TGF-ß1, reaching a concentration of 2 ng/mL of TGF-ß1 in 7 days to 14 days, which tapers subsequently. In stage-2, Dex was released linearly from 9 days to 28 days. The TD-NS group showed a significantly higher (P < 0.05) osteo/odontogenic differentiation compared to the control and free TGF-ß1 group (Free-TD) that was further corroborated with animal models/histochemical examination. The findings from this study highlighted the potential of temporal-controlled delivery of TGF-ß1 and Dex from a single nano-carrier to direct spatial and temporal-control for a cell-free tissue engineering strategy in the treatment of apical periodontitis.

Photoactivated rose bengal functionalized chitosan nanoparticles produce antibacterial/biofilm activity and stabilize dentin-collagen.

Treatment of infected teeth presents two major challenges: persistence of the bacterial-biofilm within root canals after treatment and compromised structural integrity of the dentin hard-tissue. In this study bioactive polymeric chitosan nanoparticles functionalized with rose-bengal, CSRBnp were developed to produce antibiofilm effects as well as stabilize structural-integrity by photocrosslinking dentin-collagen. CSRBnp were less toxic to fibroblasts and had significant antibacterial activity even in the presence of bovine serum albumin. CSRBnp exerted antibacterial mechanism by adhering to bacterial cell surface, permeabilizing the membrane and lysing the cells subsequent to photodynamic treatment. Photoactivated CSRBnp resulted in reduced viability of Enterococcus faecalis biofilms and disruption of biofilm structure. Incorporation of CSRBnp and photocrosslinking significantly improved resistance to degradation and mechanical strength of dentin-collagen (P<0.05). The functionalized chitosan nanoparticles provided a single-step treatment of infected root dentin by combining the properties of chitosan and that of photosensitizer to eliminate bacterial-biofilms and stabilize dentin-matrix. FROM THE CLINICAL EDITOR: In this study, bioactive polymeric chitosan nanoparticles functionalized with rose-bengal (a photosensitizer), CSRBnp were developed to produce antibiofilm effects as well as stabilize structural-integrity of dental root dentin by photocrosslinking dentin-collagen, leading to efficient elimination of bacterial-biofilms and stabilization of dentin-matrix.

Biomimetic remineralization of demineralized enamel with nano-complexes of phosphorylated chitosan and amorphous calcium phosphate.

Remineralization of enamel plays a crucial role in the progression of carious process and the management of early caries lesion. Based on the influence of phosphorylated proteins in biomineralization, the objective of this study was to synthesize nano-complexes of phosphorylated chitosan and amorphous calcium phosphate (Pchi-ACP), and evaluate their ability to remineralize enamel subsurface lesions in vitro. Pchi was synthesized using a previously established chemical method. The biomimetic remineralizing solution containing nano-complexes of Pchi-ACP was prepared by adding CaCl2 and K2HPO4 into Pchi-ACP solution (0.5 % w/v) in sequence. The final concentrations of calcium and phosphate ions were 10 and 6 mM, respectively. The nano-complexes of Pchi-ACP were characterized by Fourier-transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). During testing the enamel lesions were treated with Pchi-ACP and fluoridated remineralizing solutions, respectively. The remineralizing of enamel lesions was examined with field emission electron microscope (FE-SEM) and Micro-CT. ACP was stabilized by Pchi to form nano-complexes that were soluble in water. The size of Pchi-ACP nano-complexes particles was determined to be less than 50 nm. XRD and SAED results confirmed their amorphous phases. FE-SEM and Micro-CT results showed that the remineralizing effect of Pchi-ACP on enamel lesions was similar to that of fluoride. However, the remineralizing rate of Pchi-ACP treatment was significantly higher than that of fluoride treatment (P < 0.05). This study highlighted the potential of nanoparticles functionalized with a natural analogue involved in biomineralization, to remineralize early enamel caries.

Engineered Immunomodulatory Nanoparticles Inhibit Root Resorption and Ankylosis.

INTRODUCTION: External root resorption following avulsion injury is a complex process wherein differentiation of macrophages (Mϕ) to multinucleated osteoclasts is temporally regulated by resident periodontal fibroblasts (PDLF). The current study aims to assess the effect of engineered bioactive chitosan nanoparticles (CSNP), sustained released dexamethasone conjugated CSNP (CS-DEX) and CSNP functionalized with photosensitizer Rose Bengal (CSRB) for application in root resorption using an in-vitro PDLF-Mϕ direct coculture model and in-vivo delayed reimplantation model. METHODS: PDLF-Mϕ direct coculture system was exposed to lipopolysaccharide (LPS), macrophage colony stimulating factor, receptor activator of nuclear factor kappa ß ligand with or without CSNP/CS-DEX for 7 days. Clastic differentiation was assessed by tartrate resistant acid phosphatase (TRAP) staining on day 7. On day 2 and 7, immunofluorescence analysis was conducted to assess the expression of Mϕ polarization markers (CD80, CD206), multinucleation markers (NFATc1, STAT6) in Mϕ and matricellular protein periostin in PDLF and cytokine profiling in cell culture supernatants. Delayed replantation model with extraoral air dry/LPS exposure for 1h followed by root surface treatment with CS-DEX/CSRB was used in Wistar rats. After 21 days, rats were euthanized for histologic and immunofluorescence analysis. Statistical analysis one-way ANOVA with Tukey's multiple comparisons was used to analyze the data (P < .05). RESULTS: CS-DEX significantly reduced TRAP+ multinucleated cells and CSNP treatment showed no TRAP+ cells. Immunofluorescence analysis showed that CSNP/CS-DEX reduced CD80, NFATc1 and STAT6 expression and increased periostin as expressed by fluorescence intensity. CSNP/CS-DEX significantly reduced TNFα, MMP9 and increased IL10, TGFß1. Osteoprotegerin was upregulated only by CSNP. Root surface treatment in delayed replantation model showed that CS-DEX and CSRB substantially reduced the degree of resorption and ankylosis. Further, CD80, CD206, and MMP2 expression in groups with root surface treatment with CS-DEX and CSRB was lower than airdry/LPS group and similar to healthy control and NFATc1, STAT6, and MMP9 expressions were lower than healthy control. CONCLUSION: The engineered nanosized immunomodulatory bioactive materials chitosan nanoparticles functionalized with photosensitizer and dexamethasone effectively reduced the clastic differentiation of Mϕ in in-vitro coculture and minimized the resorption and ankylosis in a delayed reimplantation model. These biomaterials have the potential to serve as root modification agents, promoting favorable healing outcomes in cases of dental avulsion.

Deciphering 3D periodontal fibroblast-macrophage crosstalk in bioactive nanoparticle-guided immunomodulation for treating traumatic dental avulsion.

Prolonged extra-oral period in dental avulsion is often associated with loss of viability of Periodontal fibroblasts (PDLF) and increased risk of ankylosis. Root surface treatment with bioactive agents to reduce the risk of ankylosis can be a potential strategy. The objective of the study was to investigate the impact of an engineered chitosan nanoparticles (CSNP), photosensitizer Rose Bengal (RB) functionalized CSNP (CSRB) and sustained dexamethasone (CSDEX) releasing CSNP for application in management of delayed replantation of avulsed teeth. The 3D PDLF-macrophage (Mϕ) collagen model was developed and exposed to LPS, MCSF, RANKL with and without CSDEX/CSNP. Immunofluorescence and cytokine analysis was done at 2 and 7 days to assess cellular interactions. Maxillary right incisors in male Wistar rats were extracted, exposed to extraoral dry or LPS for 1 h and treated with or without CSDEX/CSRB for 1 min before replantation. Rats were euthanized after 21 days for micro-CT, TRAP, and immunofluorescence analysis. CSDEX/CSNP treatment in 3D model significantly reduced CD80, NFATc1, STAT6 and increased CD206 and periostin expression (p < 0.05). TNFα, MMP9 was downregulated and IL10, TGFß1, MMP2 upregulated with CSDEX/CSNP (p < 0.05). CSDEX/CSRB in animal study significantly reduced resorption, ankylosis, TRAP activity and osteocalcin expression and increased periostin (p<0.05). CSDEX demonstrated higher anti-inflammatory activity by downregulating TNFα, while CSNP upregulated TGFß1, periostin, and downregulated MMP9. The combination of matrix stabilization with CSRB with periostin upregulation and sustained releasing CSDEX showed potential for hampering root resorption and ankylosis in dental avulsion.

Dynamic Photoelastic Analysis of Stress Distribution in Simulated Canals Using Rotary Instruments with Varied Tip and Taper Sizes: A Quasi-3D Approach.

INTRODUCTION: To compare the stress produced on the walls of simulated canals by rotary instruments with varied tip and taper sizes. METHODS: Ninety isotropic transparent blocks, each containing a 60-degree curved canal, were distributed into 18 groups (n = 5) based on the instrument tip (sizes 10, 15, 20, 25, 30, and 35) and taper (sizes 0.02, 0.04, and 0.06). The blocks were fixed in a circular polariscope setup for dark field analysis. A digital camera was employed to capture the real-time birefringence patterns generated by each instrument. Digital image frames, corresponding to the instrument reaching the end of each canal third, were extracted and evaluated by 2 independent observers for the stress generation on canal walls. The data analysis employed a semi-quantitative scale ranging from 0 to 5. Cohen's Kappa coefficient test was used to determine the inter-observer agreement while the results were compared using Kruskal-Wallis test followed by an all-pairwise posthoc procedure (α = 5%). RESULTS: Inter-observer agreement was 0.95. A significant influence of the tip size on stress was observed across the coronal (P = .011), middle (P = .006), and apical (P = .026) thirds. In contrast, taper size did not affect the stress induced at the coronal (P = .509), middle (P = .958), or apical (P = .493) thirds. The variations in tip and taper sizes did not result in a significant stress differences among the thirds (P = .181). CONCLUSIONS: The stress significantly increased across all canal thirds with larger tip sizes of rotary instruments, whereas the taper sizes did not influence the stress when compared to the canal thirds.

Physico-chemical and biological characterization of synthetic and eggshell derived nanohydroxyapatite/carboxymethyl chitosan composites for pulp-dentin tissue engineering.

Hybrid nanohydroxyapatite/carboxymethyl chitosan (nHAp-CMC) scaffolds have garnered significant attention in the field of regenerative engineering. The current study comparatively analyzed the physicochemical and biological properties of synthetic nanohydroxyapatite (SnHA)- and eggshell-sourced nanohydroxyapatite (EnHA)- based CMC biocomposites for pulp-dentin regeneration. EnHA and CMC were synthesized through a chemical process, whereas SnHA was commercially obtained. Composite scaffolds of SnHA-CMC and EnHA-CMC (1:5 w/w) were prepared using freeze-drying method. All biomaterials were characterized by FTIR, micro-Raman, XRD, HRSEM-EDX, and TEM analyses, and their in vitro bioactivity was assessed by immersing them in simulated body fluid for 21 days. The biological properties of the composite scaffolds were evaluated by assessing cytocompatibility using MTT assay and biomineralization potential by analyzing the odontogenic gene expressions (ALP, DSPP, DMP-1 and VEGF) in human dental pulp stem cells (DPSCs) using RT-qPCR method. Characterization studies revealed that EnHA displayed higher crystallinity and superior surface morphology compared to SnHA. The composite scaffolds showed a highly interconnected porous microstructure with pore sizes ranging between 60 and 220 µm, ideal for cell seeding. All tested materials, SnHA, EnHA, and their respective composites, displayed high cytocompatibility, increased ALP activity and degree of mineralization with significant upregulation of odontogenic-related genes on DPSCs (p < 0.05). Nevertheless, the odontogenic differentiation potential of EnHA-CMC on DPSCs was significantly higher when compared to SnHA-CMC. The findings from this study highlight the potential of EnHA-CMC as a promising candidate for pulp-dentin engineering.

Impact of Type 2 Diabetes Mellitus on the Occurrence of Vertical Root Fracture: A Case Control Study.

INTRODUCTION: The aim of this case-control study was to examine the relationship between type 2 diabetes mellitus (DM) and the occurrence of VRFs. The crack extension, dentin sclerosis, and chemical characteristics of root dentin in teeth with VRF from patients with/without DM were also compared. METHODS: One hundred and thirty-two patients diagnosed with VRF in crowned root filled posterior teeth were selected. The study was conducted in 2 parts. In Part-1: The cases were matched with control teeth (1:1) for age (±5 years), sex, tooth type, apical extent of root filling, time period after root filling to a diagnosis of VRF, presence or absence of intracanal post and abutment status. The presence or absence of type 2 DM (HbA1c > 6.5) was recorded. In Part-2: The extracted teeth with VRF from the case control study were used to evaluate the extension of VRF, presence of sclerotic dentin and isthmus using a microscopic analysis; while the levels of pentosidine, collagen cross-linking ratio and mineral-collagen ratio were determined by Fourier-transform infrared spectroscopy. The distribution of DM between cases and controls was analyzed using Pearson Chi-Square test and Odds Ratio estimated. Chemical composition data was analyzed using Mann-Whitney test. The extent of sclerotic dentin was analyzed using Pearson Chi-Square test. RESULTS: When compared to patients without DM, patients with DM had 2.67 (95% CI: 1.6-4.45) folds higher odds for occurrence of VRF. Pentosidine (P = .014), collagen cross-linking ratio(P = .047), mineral-collagen ratio (P = .009) and sclerotic dentin extent (P = .0009) were significantly higher in patients with DM and VRF. CONCLUSIONS: Type 2 DM was more often associated with VRFs in root canal treated teeth with crowns. Root dentin from patients with type 2 DM and VRF had higher levels of pentosidine, collagen cross-linking ratio, mineral to collagen ratio and sclerotic dentin.

Pericervical Dentin Metrics in Mandibular First Molars Determined with Digital Periapical Radiography and Cone-beam Computed Tomography.

INTRODUCTION: Pericervical root dentin is decisive for the long-term mechanical integrity of root-filled teeth. Current treatment protocol does not include a customized step to determine the pretreatment residual pericervical root dentin. OBJECTIVE: To determine and compare the residual root dentin and canal width using digital periapical radiography (DPR) and cone-beam computed tomography (CBCT) at the apical limit of the pericervical area (PCA) in mandibular first molars. METHODS: DPR and CBCT images of 60 patients with age between 22 and 76 years were used to determine (a) the mesiodistal widths of the root canal (pericervical dimensions [PCL]-C) and the root (PCL-R) of mandibular first molars at the apical limit of the PCA and (b) the intracanal distance from the apical limit of the PCA to the radiographic apex (intracanal distance [ICD]). The correlation between the PCL and ICD measurements obtained from DPR and CBCT were evaluated. RESULTS: Values between 0.10-0.80 mm and 0.00-1.10 mm were obtained for PCL-C using DPR and CBCT respectively (95% CI). The PCL values between 0.90-2.30 mm and 0.00-2.30 mm were obtained from DPR and CBCT respectively (95% CI). The ICD ranged between 4.6-12.3 mm in DPR and 4.40-12.0 mm in CBCT (95% CI). The comparative analysis showed differences from -0.9 to 0.5 mms for PCL and -2.00 to 1.5 mms for ICD between DPR and CBCT techniques respectively. CONCLUSION: The PCL and ICD determined from DPR and CBCT provided the pericervical dentin metrics that could be utilized clinically as a guideline for decision-making in endodontic treatment.

Nociceptor-Macrophage Interactions in Apical Periodontitis: How Biomolecules Link Inflammation with Pain.

Periradicular tissues have a rich supply of peripheral afferent neurons, also known as nociceptive neurons, originating from the trigeminal nerve. While their primary function is to relay pain signals to the brain, these are known to be involved in modulating innate and adaptive immunity by initiating neurogenic inflammation (NI). Studies have investigated neuroanatomy and measured the levels of biomolecules such as cytokines and neuropeptides in human saliva, gingival crevicular fluid, or blood/serum samples in apical periodontitis (AP) to validate the possible role of trigeminal nociceptors in inflammation and tissue regeneration. However, the contributions of nociceptors and the mechanisms involved in the neuro-immune interactions in AP are not fully understood. This narrative review addresses the complex biomolecular interactions of trigeminal nociceptors with macrophages, the effector cells of the innate immune system, in the clinical manifestations of AP.

Periodontal Fibroblasts-Macrophage Crosstalk in External Inflammatory Root Resorption.

INTRODUCTION: This study aimed to understand the influence of periodontal fibroblasts (PDLFs) on clastic differentiation of macrophages (Mφ) in different resorptive environments. METHODS: PDLF-Mφ direct coculture (juxtacrine) was seeded on dentin, cementum, and polystyrene with/without lipopolysaccharide, macrophage colony-stimulating factor, and receptor activator of nuclear factor kappa beta ligand for 7 and 14 days and stained for tartrate-resistant acid phosphatase (TRAP) activity. PDLF-Mφ cocultured on polystyrene were immunostained for CD80, CD206, NFATc1, STAT6, and periostin, and cell culture supernatants were assessed for cytokines on days 2 and 7. Mφ grown in conditioned media of PDLFs (paracrine) and Mφ monoculture were used as controls. Data was analyzed using Student t test and one-way analysis of variance with the Tukey multiple comparisons test (P < .05). RESULTS: PDLF-Mφ coculture showed a higher number of TRAP-positive multinucleated cells than Mφ monoculture on dentin and polystyrene. No TRAP-positive multinucleated cells were observed in paracrine and cementum. The expression of CD80 and CD206 in PDLF-Mφ was similar at day 2, whereas CD206 was greater than CD80 at day 7. The expression of STAT6 was greater than NFATc1 at both days 2 and 7 (P < .05). Periostin expression in the presence of the lipopolysaccharide, macrophage colony-stimulating factor, and receptor activator of nuclear factor kappa beta ligand combination was down-regulated in PDLF monoculture, whereas it was up-regulated in PDLF-Mφ coculture. The cytokine profile of PDLF-Mφ on day 2 was predominated by interleukin (IL)-1ß, tumor necrosis factor alpha, and MMP9 and MMP2 on day 7. IL-6 and IL-8 showed steady expression at both days 2 and 7. CONCLUSIONS: The study highlights the juxtacrine effect of PDLFs on the clastic differentiation of Mφ with a difference in clastic activity between dentin and cementum. The study also emphasizes the temporal effect of tumor necrosis factor alpha, MMP2, MMP9, and IL-1ß on intercellular crosstalk in resorptive environments.

Synchronized Microbubble Photodynamic Activation to Disinfect Minimally Prepared Root Canals.

INTRODUCTION: The purpose of this study was to evaluate the antimicrobial efficacy of a novel irrigation strategy using synchronized microbubble photodynamic activation (SYMPA) in a minimally prepared single canal. METHODS: Single-canal mandibular incisors were inoculated with Enterococcus faecalis for 3 weeks and randomly allocated to 4 groups based on the irrigation protocols: (1) control (saline), (2) conventional needle irrigation (CI), (3) ultrasonic-assisted irrigation (UI), and (4) irrigation with SYMPA. The first 3 groups were instrumented to size 25.07v (WaveOne Gold Primary; Dentsply Sirona, Johnson City, TN), and the SYMPA group was minimally prepared to size 20.07v (WaveOne Gold Small, Dentsply Sirona). The apical 5 mm was resected for microbiological assessment using the culture technique (colony-forming unit), adenosine-5'-triphosphate-based viability assay (relative luminescence units), and the percentage of live bacteria using confocal laser scanning microscopy. RESULTS: Log colony-forming units from the UI (2.37 ± 0.66) and SYMPA (2.21 ± 0.86) groups showed a reduction compared with the control (5.16 ± 0.75) and CI (4.08 ± 1.19) groups. Relative luminescence unit reduction was significant for UI (619.08 ± 352.78) and SYMPA (415.25 ± 329.51) compared with the control (1213.2 ± 880.03) (P < .05). The percentage of live bacteria was significantly lower in the UI and SYMPA groups compared with the control and CI groups. Although higher microbial reduction was observed in SYMPA compared with UI, there was no statistical significance (P > .05). CONCLUSION: SYMPA in minimally prepared canals showed significant antimicrobial efficacy. The novel irrigation strategy using SYMPA could be an effective disinfection strategy for minimally prepared root canals.