Effect of magnesium oxide nanoparticles and LED irradiation on the viability and differentiation of human stem cells of the apical papilla.

PURPOSE: Currently, regenerative endodontic treatments are gaining more and more attention, and stem cells play a significant role in these treatments. In order to enhance stem cell proliferation and differentiation, a variety of methods and materials have been used. The purpose of this study was to determine the effects of magnesium oxide nanoparticles and LED irradiation on the survival and differentiation of human stem cells from apical papilla. METHODS: The MTT test was used to measure the cell survival of SCAPs that had been exposed to different concentrations of magnesium oxide nanoparticles after 24 and 48 h, and the concentration with the highest cell survival rate was picked for further studies. The cells were classified into four distinct groups based on their treatment: (1) control, which received no exposure, (2) exposure to magnesium oxide nanoparticles, (3) exposure to light emitting diode (LED) irradiation (635 nm, 200 mW/cm2) for 30 s, (4) exposure simultaneously with magnesium oxide nanoparticles and LED irradiation. A green approach was employed to synthesize magnesium oxide nanoparticles. Quantitative real time PCR was used to measure the gene expression of osteo/odontogenic markers such as BSP, DSPP, ALP and DMP1 in all four groups after treatment, and Alizarin red S staining (ARS) was used to determine the osteogenic differentiation of SCAPs by demonstrating the Matrix mineralization. RESULTS: The highest viability of SCAPs was observed after 24 h in concentration 1 and 10 µg/mL and after 48 h in concentration 1 µg/mL, which were not significantly different from the control group. In both times, the survival of SCAPs decreased with increasing concentration of magnesium oxide nanoparticles (MgONPs). According to the results of Real-time PCR, after 24 and 48 h, the highest differentiation of BSP, DMP1, ALP and DSPP genes was observed in the LED + MgONPs group, followed by MgONPs and then LED, and in all 3 experimental groups, it was significantly higher than control group (P < 0.05). Also, after 24 and 48 h, the density of ARS increased in all groups compared to the control group, and the highest density was observed in the MgONPs + LED and MgONPs groups. CONCLUSION: This research concluded that exposure to SCAPs, MgONPs, and LED irradiation has a significant effect on enhancing gene expression of odontogenic/osteogenic markers and increasing matrix mineralization.

Deep learning for determining the difficulty of endodontic treatment: a pilot study.

BACKGROUND: To develop and validate a deep learning model for automated assessment of endodontic case difficulty from periapical radiographs. METHODS: A dataset of 1,386 periapical radiographs was compiled from two clinical sites. Two dentists and two endodontists annotated the radiographs for difficulty using the "simple assessment" criteria from the American Association of Endodontists' case difficulty assessment form in the Endocase application. A classification task labeled cases as "easy" or "hard", while regression predicted overall difficulty scores. Convolutional neural networks (i.e. VGG16, ResNet18, ResNet50, ResNext50, and Inception v2) were used, with a baseline model trained via transfer learning from ImageNet weights. Other models was pre-trained using self-supervised contrastive learning (i.e. BYOL, SimCLR, MoCo, and DINO) on 20,295 unlabeled dental radiographs to learn representation without manual labels. Both models were evaluated using 10-fold cross-validation, with performance compared to seven human examiners (three general dentists and four endodontists) on a hold-out test set. RESULTS: The baseline VGG16 model attained 87.62% accuracy in classifying difficulty. Self-supervised pretraining did not improve performance. Regression predicted scores with ± 3.21 score error. All models outperformed human raters, with poor inter-examiner reliability. CONCLUSION: This pilot study demonstrated the feasibility of automated endodontic difficulty assessment via deep learning models.

The effects of melatonin on the viability and osteogenic/odontogenic differentiation of human stem cells from the apical papilla.

BACKGROUND: An experimental study was conducted to examine whether melatonin influences osteogenic/odontogenic differentiation of human stem cells derived from the apical papilla (hSCAPs). MATERIALS AND METHODS: In order to isolate hSCAPs, the undeveloped root of a third molar of a human tooth was used. Melatonin was administered to the experimental groups in an osteogenic medium. No treatment was administered to the control group. The methyl thiazolyl tetrazolium (MTT) assay was performed on days 1, 2, and 3 to assess cell viability (n = 8). A determination of odontogenic/osteogenic differentiation was accomplished using alkaline phosphatase (ALP) activity alizarin red staining (ARS) (n = 6), and the expression of osteogenic genes by real-time polymerase chain reaction (RT-PCR) (n = 3) on days 1, 2, and 7. Evaluation of the data was conducted using SPSS version 18. All experiments were conducted at least three times. The Mann Whitney U test, the ANOVA analysis, Tukey's test, and t-test was implemented to analyze the data (α = 0.05). RESULTS: After 24 h, 48 h, and 72 h, No significant difference was observed between the control group and the melatonin treatment group in terms of viability of hSCAPs. (from 1 up to 10 µg/ml) (P > 0.05). The assessment of ARS and ALP activity showed that melatonin treatment enhanced osteogenic differentiation of hSCAPs (P < 0.001). Melatonin treatment caused hSCAPs to show an increase of genes related to osteogenic/odontogenic differentiation. These genes included ALP, dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), and bone sialoprotein (BSP) (P < 0.001). CONCLUSIONS: Melatonin treatment enhanced osteogenic/odontogenic differentiation of hSCAPs with a dose dependent effect on cell viability.

Effects of CEM cement and emdogain on proliferation and differentiation of human stem cells from the apical papilla: a comparative in vitro study.

OBJECTIVES: This study compared the effects of calcium-enriched mixture (CEM) cement, Emdogain (EMD), and their combination (CEM/Emdogain) on the differentiation and proliferation of stem cells from the apical papilla (SCAPs). METHODS: In this in vitro, experimental study, SCAPs were isolated from two sound immature impacted third molars and cultured. After ensuring their stemness by detecting cell surface markers they were exposed to CEM cement, Emdogain, and CEM cement coated with Emdogain for 24 and 72 h. The control cells did not undergo any intervention. Cell viability [by methyl thiazolyl tetrazolium (MTT) assay], expression of odontogenic differentiation genes [by quantitative reverse-transcription polymerase chain reaction (qRT-PCR)], and alkaline phosphatase (ALP) activity (by ALP staining kit) were evaluated. Data were analyzed by one-way ANOVA, t-test, and Mann-Whitney test (α = 0.05). RESULTS: Cell viability in the CEM cement and CEM/Emdogain groups decreased compared with the control group at 72 h (P < 0.05). Expression of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), bone sialoprotein (BSP) genes, and ALP activity significantly increased in all three experimental groups compared with the control group at both 24 and 72 h. This increase was substantially more significant in CEM/Emdogain group (P > 0.05). The number of mineralized nodules significantly increased in all groups at 72 h, with a higher rate in the CEM/Emdogain group. CONCLUSION: All biomaterials increased the differentiation of SCAPs, expression of odontogenic differentiation genes, and ALP activity, but CEM/Emdogain was considerably more effective for this purpose.

Association of root morphology of mandibular second molars on panoramic-like and axial views of cone-beam computed tomography.

BACKGROUND: Knowledge about the anatomy and morphology of the root canal system is essential for successful surgical and non-surgical root canal treatments. However, precise assessment of the root morphology and anatomy is not often possible on two-dimensional radiographs. This study aimed to investigate the association of root morphology of mandibular second molars on panoramic-like and axial views of cone-beam computed tomography (CBCT). METHODS: This cross-sectional study evaluated 1,231 CBCT scans of mandibular second molars obtained between October 2018 and February 2022 that were retrieved from the archives of a private radiology clinic. Panoramic-like images were reconstructed from the CBCT scans. The root morphology of mandibular second molars was classified on panoramic-like images as type 1, 2, 3, 4, or 5. The root pattern on axial CBCT images was classified into three types of single, double and C-shaped. The association of root morphology on panoramic-like and axial CBCT views was analyzed by the Chi-square test and Fisher's exact test at 0.05 level of significance. RESULTS: Of all, 62.7% of mandibular second molars were type 1; out of which, 97.3% had a double-root pattern on axial CBCT images. Also, 28.6% of them were type 2; of which, 92.6% had a double-root pattern. Moreover, 3.9% were type 3; of which, 47.9% had a C-shaped pattern; 0.9% were type 4, and 45.5% of them showed a single-root pattern; 3.8% were type 5 with 76.6% of them showing a single-root pattern. The prevalence of C-shaped canals was higher in females, and most C-shaped canals had a C3 pattern. CONCLUSION: Root morphology on panoramic-like CBCT views had a strong association with the root canal pattern on axial CBCT views. According to the results, mandibular second molars with a type 3 morphology on panoramic-like CBCT images are highly probable to have a C-shaped canal.

Effect of copper oxide nanoparticles and light-emitting diode irradiation on the cell viability and osteogenic/odontogenic differentiation of human stem cells from the apical papilla.

OBJECTIVES: This experimental study aimed to assess the effect of copper oxide nanoparticles (CuONPs) and light-emitting diode (LED) irradiation on the cell viability and osteogenic/odontogenic differentiation of human SCAPs. METHODS: After the culture of SCAPs, the effects of different concentrations of CuONPs on cell viability were evaluated by the methyl thiazolyl tetrazolium (MTT) assay after 24 and 48 h, and the optimal concentration was determined (n = 12). SCAPs were then divided into four groups based on the type of treatment: (I) no-treatment control group, (II) exposure to CuONPs, (III) LED irradiation (635 nm, 200 mW/cm2) for 30 s, and (IV) exposure to CuONPs combined with LED irradiation. CuONPs were synthesized by a green technique, which was based on reduction and simultaneous stability of copper ions by using the pomegranate peel extract. After treatments, the expression of osteogenic/odontogenic markers including dentin sialophosphoprotein (DSPP), bone sialoprotein (BSP), alkaline phosphatase (ALP), and dentin matrix acidic phosphoprotein 1 (DMP1) was evaluated in all four groups using quantitative real-time polymerase chain reaction (PCR) (n = 16). Also, osteogenic differentiation of SCAPs was evaluated qualitatively by alizarin red staining (ARS) to assess the matrix mineralization (n = 4). SPSS version 18 was used for data evaluation. The Kruskal-Wallis and Mann-Whitney tests were used to compare the groups. RESULTS: Exposure to 1 µg/mL CuONPs resulted in maximum viability of SCAPs. Concentrations of CuONPs over 10 µg/mL significantly decreased the viability of SCAPs. Real-time PCR showed that the expression of DMP1, BSP, ALP, and DSPP in CuONPs + LED and LED groups was significantly higher than that in CuONPs and control groups at both 24 and 48 h (P < 0.05). The density of ARS increased in all experimental groups after 24 h, and in CuONPs + LED and CuONPs groups after 48 h, compared to the control group. CONCLUSION: Addition of CuONPs and LED irradiation of SCAPs in the culture medium significantly enhanced their osteogenic/odontogenic differentiation.

The effect of low-dose aspirin on aspirin triggered lipoxin, interleukin 1 beta, and prostaglandin E2 levels in periapical fluid: a double-blind randomized clinical trial.

BACKGROUND: The role of pro-resolving mediators in inflammation is a new concern in research. The effect of low-dose aspirin on production of a special kind of these mediators named aspirin triggered lipoxin (ATL) has been studied on different tissues. This randomized clinical trial evaluated the effect of low-dose aspirin on ATL and pro-inflammatory mediators' level in periapical fluid of necrotic teeth with large lesions. METHODS: Twenty-four patients with necrotic pulp and periapical lesion were randomly assigned to low-dose aspirin and placebo groups. In the first appointment, canals were shaped up to F3 size and #40 K-file and cleaned with 10 milliliters 2.5% sodium hypochlorite and 17% Ethylenediaminetetraacetic acid. Periapical fluid was sampled by a paper cone. The tooth was temporized without any intracanal medication. Tablets were administered for 7 days, then the teeth were re-opened and the sampling were repeated. Interleukin-1 beta (IL-1ß), prostaglandin E2 (PGE2) and ATL were analyzed by enzyme-linked immunosorbent assay. Data were analyzed with paired t-test using SPSS statistical software, version 21 (α = 0.05). RESULTS: A significant reduction in PGE2 and IL-1ß was noted in the aspirin-treated group while an increase in ATL was observed (P < 0.001). There was no significant difference in the mediator scores before and after in the placebo-treated group (P > 0.05). CONCLUSION: Low-dose aspirin can influence the inflammatory process by reducing pro-inflammatory mediators such as PGE2 and IL-1ß, as well as increasing the pro-resolving mediators such as ATL. TRIAL REGISTRATION: IRCT20191211045702N1.

Effect of biodentine coated with emdogain on proliferation and differentiation of human stem cells from the apical papilla.

BACKGROUND: This study assessed the effect of Biodentine coated with Emdogain (Biodentine/Emdogain) on proliferation and differentiation of human stem cells from the apical papilla (SCAPs). METHODS AND RESULTS: In this in vitro, experimental study, SCAPs were isolated from two immature impacted third molars and cultured. After ensuring the stemness of the cells by assessing the cell surface markers, they were exposed to Biodentine, Emdogain, and Biodentine/Emdogain for 24 and 72 h. The control cells did not receive any intervention. Cell viability was evaluated by the methyl thiazolyl tetrazolium assay. Expression of odontogenic differentiation genes was analyzed by the quantitative reverse transcription polymerase chain reaction. Alkaline phosphatase (ALP) activity was quantified by the respective kit. Data were analyzed by one-way ANOVA, t-test, and Mann-Whitney test (α = 0.05). Cell viability did not change after 24 h of exposure to biomaterials. At 72 h, the viability of the cells exposed to Biodentine and Biodentine/Emdogain decreased compared with the control group. The expression of dentin sialophosphoprotein, dentin matrix protein 1, and bone sialoprotein genes, and ALP activity significantly increased in all three experimental groups, compared with the control group at both 24 and 72 h; this increase was significantly greater in Biodentine/Emdogain group. The number of mineralized nodules significantly increased in all groups after 72 h with a greater rate in Biodentine/Emdogain group. CONCLUSIONS: All biomaterials increased the differentiation of SCAPs, expression of odontogenic genes, and ALP activity, but Biodentine/Emdogain was significantly more effective for this purpose.

Effect of diode low level laser and red light emitting diode irradiation on cell proliferation and osteogenic/odontogenic differentiation of stem cells from the apical papilla.

BACKGROUND: This experimental study aimed to assess the effect of irradiation of red light-emitting diode (LED) and Diode low-level laser (LLL) on osteogenic/odontogenic differentiation of stem cells from the apical papilla (SCAPs). MATERIALS AND METHODS: SCAPs were isolated from the human tooth root. The experimental groups were subjected to 4 J/cm2 diode low level laser and red LED irradiation in osteogenic medium. The control group did not receive any irradiation. Cell viability/proliferation of SCAPs was assessed by the methyl thiazolyl tetrazolium (MTT) assay on days 1 and 2 (n = 9). Osteogenic differentiation was evaluated by alizarin red staining (ARS) (n = 3), and expression of osteogenic genes by real-time polymerase chain reaction (RT-PCR) (n = 12) on days 1 and 2. SPSS version 18 was used for data evaluation. The Kruskal-Wallis and Mann-Whitney tests were used to compare the groups at each time point. RESULTS: The MTT assay showed no significant difference in cell viability/proliferation of SCAPs in the low level laser, red LED, and control groups at 24 or 48 h (P < 0.001). The ARS assessment showed that low level laser and red LED irradiation enhanced osteogenic differentiation of SCAPs. low level laser and red LED irradiation both induced over-expression of osteogenic/dentinogenic genes including alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), and bone sialoprotein (BSP) in SCAPs. Up-regulation of genes was significantly greater in low level laser irradiation group than red LED group (P < 0.001). CONCLUSION: Diode low level laser irradiation with 4 J/cm2 energy density and red LED irradiation enhanced osteogenic differentiation of SCAPs without adversely affecting cell viability.

Cone-beam computed tomographic analysis of apical transportation and centering ratio of ProTaper and XP-endo Shaper NiTi rotary systems in curved canals: an in vitro study.

BACKGROUND: Cleaning and shaping of the root canal system is an important step of endodontic treatment. Canal transportation is a common procedural error in preparation of curved canals. This study aimed to compare the canal transportation and centering ratio of two rotary files in curved canals using cone-beam computed tomography (CBCT). METHODS: Forty-four extracted human mandibular first molars with mature apices and 10° to 30° apical curvature were selected. The samples were randomly divided into two groups (n = 22) with similar curvature. The canals were prepared with ProTaper and XP-endo Shaper file systems according to the manufacturers' instructions. The CBCT images were obtained using Cranex 3D CBCT scanner before and after root canal preparation, and canal transportation and centering ratio of the files at 3, 4 and 5 mm levels from the apex were calculated. Data were compared between the two groups using independent t-test at 0.05 level of significance. RESULTS: The ProTaper Universal caused greater canal transportation and had lower centering ratio than XP-endo Shaper in both mesiodistal and buccolingual directions at all levels from the apex. The difference between the two groups regarding canal transportation was significant at all levels from the apex in buccolingual direction (P < 0.05) except for 3 mm from the apex (P > 0.05). The difference between the two groups regarding centering ratio was not significant (P > 0.05) in mesiodistal direction at all levels except for 4 mm from the apex (P < 0.05). CONCLUSION: The ProTaper Universal causes greater canal transportation in both buccolingual and mesiodistal directions than XP-endo Shaper.