Effect of Low-Level Diode Laser and Red Light-Emitting Diode on Survival and Osteogenic/Odontogenic Differentiation of Human Dental Pulp Stem Cells.

Background: This investigation set out to compare the impacts of low-level diode laser (LLDL) and red light-emitting diode (LED) on the survival of human dental pulp stem cells (hDPSCs) and osteogenic/odontogenic differentiation. Methods and materials: In this ex vivo experimental study, the experimental groups underwent the irradiation of LLDL (4 J/cm2 energy density) and red LED in the osteogenic medium. Survival of hDPSCs was assessed after 24 and 48 h (n = 9) using the methyl thiazolyl tetrazolium (MTT) assay. The assessment of osteogenic/odontogenic differentiation was conducted using alizarin red staining (ARS; three repetitions). The investigation of osteogenic and odontogenic gene expression was performed at two time points, specifically 24 and 48 h (n = 12). This analysis was performed utilizing real-time reverse-transcription polymerase chain reaction (RT-PCR). The groups were compared at each time point using SPSS version 24. To analyze the data, the Mann-Whitney U test, analysis of variance, Tukey's test, and t-test were utilized. Results: The MTT assay showed that LLDL significantly decreased the survival of hDPSCs after 48 h, compared with other groups (p < 0.05). The qualitative results of ARS revealed that LLDL and red LED increased the osteogenic differentiation of hDPSCs. LLDL and red LED both upregulated the expression of osteogenic/odontogenic genes, including bone sialoprotein (BSP), alkaline phosphatase (ALP), dentin matrix protein 1 (DMP1), and dentin sialophosphoprotein (DSPP), in hDPSCs. The LLDL group exhibited a higher level of gene upregulation (p < 0.0001). Conclusions: The cell survival of hDPSCs was reduced, despite an increase in osteogenic/odontogenic activity. Clinical relevance: Introduction of noninvasive methods in regenerative endodontic treatments.

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.

Effect of 810 nm Diode Laser Irradiation on the Time of Initiation and Depth of Anesthesia for Endodontic Treatment of Mandibular First Molars with Symptomatic Irreversible Pulpitis: A Clinical Trial.

Objective: In endodontic treatments, performing appropriate anesthesia in patients with irreversible pulpitis in mandibular molars may result in pain and severe problems. The irradiation of low-level lasers could be effective in this regard due to its anti-inflammatory and regenerative properties. This study aimed to assess the effect of 810 nm diode laser on the time of initiation and depth of anesthesia for endodontic treatment of mandibular first molars with symptomatic irreversible pulpitis. Materials and methods: This randomized controlled clinical trial evaluated 60 patients requiring endodontic treatment of mandibular first molars with symptomatic irreversible pulpitis and pain score ≥114 according to the Heft-Parker visual analog scale (HP-VAS). The teeth were randomized into two groups of diode laser and control. In the diode laser group, 810 nm diode laser with 300 mW power and 15 J/cm2 energy density was irradiated to the buccal surface of tooth crowns for 20 sec at 2 mm distance immediately before anesthesia administration. Laser in off mode was used in the control group. Inferior alveolar nerve block was then performed using 2% lidocaine with 1:80,000 epinephrine. After anesthetic injection, the mandibular first molar and canine teeth (control) were tested by an electric pulp tester every 2 min. Two consecutive negative responses to 80 mA indicated the initiation of anesthesia. HP-VAS forms were filled out by patients to assess their level of pain during the procedure. Data were analyzed by the Student's t and Chi-square tests, and analysis of variance (α = 0.05). Results: No remarkable difference was noted between the laser group and control groups in pain severity or anesthesia onset (p > 0.05). Conclusions: Low-level (810 nm) diode laser did not affect the time of initiation or depth of anesthesia in endodontic treatment of mandibular first molars with symptomatic irreversible pulpitis. Clinical trials registration: Iranian Registry of Clinical Trials (IRCT20181222042076N1).

Cone-beam computed tomography assessment of the root canal morphology of primary molars.

BACKGROUND: This study aimed to assess the root canal morphology of primary molars using cone-beam computed tomography (CBCT). METHODS: This cross-sectional study evaluated 60 maxillary and mandibular primary first and second molars on CBCT scans of patients retrieved from the archives of Hamadan School of Dentistry between 2018-2020. The teeth were evaluated regarding the number of roots and canals, canal type according to the Vertucci's classification, and root surface concavities. Data were analyzed descriptively and by independent t-test. RESULTS: The most frequent number of canals and roots in the maxillary right and left first molars was 3 canals (60%) and 3 roots (80%). These values were 4 canals (80%) and 5 canals (50%) with 3 roots in the maxillary right and left second molars, respectively, 4 canals (100%) and 2 roots (50%), and 3 canals (60%) and 2 roots (50%) in mandibular right and left first molars, respectively, and 4 canals (92.3%) and 3 roots (61.5%) in mandibular right and left second molars. Vertucci's type IV was the most common canal type in mesial and distal canals, type I was the most common in mesiobuccal, mesiolingual, distobuccal, and distolingual, and types I and II were the most common in the palatal canal. The maximum and minimum concavities were noted in the buccal (26.7%) and mesial (8.3%) surfaces, respectively. CONCLUSIONS: A wide variation exists in the number of roots and canals of maxillary and mandibular primary molars, which calls for further attention in treatment of such teeth.

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.

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.

Comparative effects of Er:YAG laser, and EDTA, MTAD, and QMix irrigants on adhesion of stem cells from the apical papilla to dentin: A scanning electron microscopic study.

Background: Dentin conditioning can affect the adhesion of stem cells in endodontic regenerative treatments. This study aimed to assess the effects of the most commonly used endodontic irrigants, namely, ethylenediaminetetraacetic acid (EDTA), MTAD, and QMix in comparison with Er:YAG laser (as a novel modality for root canal disinfection) on the adhesion of stem cells from the apical papilla (SCAPs) to dentin. Material and Methods: Forty dentin specimens were prepared and subjected to different treatments in 5 groups (n=8) of control, irrigation with EDTA for 1 minute, irrigation with MTAD for 5 minutes, irrigation with QMix for 5 minutes, and Er:YAG laser irradiation. SCAPs were isolated from third molar tooth buds that two-thirds of their roots had formed. The cells were cultured on dentin specimens for 3 days and were counted using scanning electron microscopy (SEM). Results: MTAD resulted in significantly lower adhesion of cells to dentin compared with other groups (P<0.05). All other modalities induced cell adhesion with no significant difference with each other (P>0.05). Conclusions: Despite many favorable properties, MTAD cannot serve as an optimal irrigant in endodontic regenerative procedures since it inhibits the adhesion of SCAPs to dentin and impairs an important step in tissue engineering. Key words:Endodontic Regeneration, Er-YAG laser, MTAD, QMix, EDTA, SCAP, stem cell adhesion.

Comparison of the Antimicrobial Efficacy of Calcium Hydroxide and Photodynamic Therapy Against Enterococcus faecalis and Candida albicans in Teeth With Periapical Lesions; An In Vivo Study.

Introduction: Elimination of pathological microflora of root canal systems is a major goal in endodontic treatment. This study aimed to compare the antimicrobial efficacy of calcium hydroxide as an intracanal medication and antibacterial photodynamic therapy (aPDT) against Enterococcus faecalis and Candida albicans in teeth with periapical (PA) lesions. Methods: This in vivo study was conducted on 20 patients with single-rooted mandibular premolar with previously failed endodontic treatment. This study was performed as a clinical trial (IRCTID: IRCT2016090429686N1). After conventional chemo-mechanical root canal preparation (hand and rotary instruments and 2.5% NaOCl), microbiological samples were obtained using sterile paper points, then stored in thioglycolate solution and transferred to a microbiology laboratory. Group 1 (n = 10) specimens underwent aPDT (diode laser 808 nm + 50 mg/mL methylene blue), while creamy calcium hydroxide paste was used in group 2 for a duration of 1 week. A control sample was taken with sterile paper points and F3 Protaper rotary file. The samples were dispersed in transport medium, serially diluted, and cultured on selective mediums to determine the number of colony forming units (CFUs). Data were analyzed by Mann-Whitney U test at 5% significance level. The significance level for all analyses was set at P < 0.05. Results: Number of CFU significantly decreased in both groups after the interventions (P < 0.001); however, there was no significant difference in the colony count between the 2 groups. Conclusion: aPDT and calcium hydroxide therapy showed the same antimicrobial efficacy on E. faecalis and C. albicans.

Vital Pulp Therapy with Calcium-Silicate Cements: Report of Two Cases.

This article describes successful use of calcium-enriched mixture (CEM) cement and Biodentine in apexogenesis treatment in two 8-year-old patients, one with immature permanent molar diagnosed primarily with irreversible pulpitis and the other with partially vital maxillary central incisor. After access cavity preparation, partial pulpotomy in molar and full pulpotomy in central was performed, and the remaining pulps was capped with either Biodentine or CEM cement, in each tooth. The crowns were restored with composite filling material at the following visit. The post-operative radiographic and clinical examinations (approx. average of 16 months) showed that both treated teeth remained functional, with complete root development and apex formation. A calcified bridge was produced underneath the capping material. No further endodontic intervention was necessary. Considering the healing potential of immature vital pulps, the use of CEM cement and Biodentine for apexogenesis might be an applicable choice. These new endodontic biomaterials might be appropriate for vital pulp therapies in an immature tooth. However, further clinical studies with longer follow-up periods are recommended.

A comparison between effect of photodynamic therapy by LED and calcium hydroxide therapy for root canal disinfection against Enterococcus faecalis: A randomized controlled trial.

BACKGROUND: Insufficient root canal disinfection is one of the main reasons for persistent periapical pathology. Photodynamic therapy (PDT) has been proven effective in disinfecting infected root canals. The aim of this study was to evaluate the antimicrobial effect of photo activated disinfection (PAD) when using toluidine blue as photosensitizer and a LED lamp after the conventional treatment, and comparing it with calcium hydroxide therapy in vivo. METHODS: This clinical trial includes 20 patients with molars requiring endodontic retreatment. After the conventional treatment, first microbiological samples were obtained using sterile rotary ProTaper F2 file and 3 paper points and transferred to a microbiology laboratory. Group 1 (n=10) specimens underwent PAD with photosensitizer (PS) solution (0.1mg/mL TB) and irradiation with Fotosan light emitting diode (LED) lamp (635nm, 200mW/cm2) for 60s. Creamy Ca(OH)2 paste was used in group 2 (n=10) for two weeks. A second sample was then obtained. The samples were cultured and then bacterial colonies were counted. Data included number of colony forming units (CFUs) before and after treatments, analyzed by t-test and analysis of covariance (ANCOVA) using SPSS vs.18. RESULTS: A significant difference between results of before and after treatment of both groups (calcium hydroxide therapy p=0.02<0.05, PAD p<0.0001) indicated the efficacy of both treatments. The mean numbers for log 10CFUs/mL before calcium hydroxide therapy and PAD with LED irradiation was 10.1968 and 11.3773. After treatment, the mean numbers were 9.4202 and 8.3772, respectively. The difference in results after treatment between groups was significant (p=0.01<0.05) and indicate that PAD was more effective. CONCLUSION: PAD and calcium hydroxide therapy, as auxiliary methods adjunct to conventional root canal therapy, are both effective in root canal disinfection. In comparison with calcium hydroxide therapy, PAD leads to a greater reduction in enterococcus faecalis number in the infected root canals.

Comparative Evaluation of the Efficacy of Hand-Wrist and Cervical Vertebrae Radiography for the Determination of Skeletal Age.

BACKGROUND: Prediction of skeletal growth is necessary for growth modification and surgical orthodontic treatments and is usually done by assessing skeletal maturity indicators in hand-wrist radiographs. The use of growth stages of cervical vertebrae in lateral cephalograms has been suggested to avoid overexposure. OBJECTIVES: This study seeks to assess the degree of agreement between hand-wrist and cervical vertebrae maturation stages for skeletal age determination and prediction of the peak growth spurt (PGS). PATIENTS AND METHODS: This cross-sectional study was conducted with 67 boys and 66 girls between 8 and 18 years of age, divided into 11 age groups; 266 hand-wrist radiographs and lateral cephalograms were obtained and analyzed. Hand-wrist maturation stages were evaluated according to the Grave and Brown, Bjork system (stages 1 - 9). The cervical vertebral maturation stage (CVMS) was determined on lateral cephalograms based on a system described by Baccetti et al. (CVMS 1-5). To apply the Cohen's kappa index, the stages of growth were reduced to 5 intervals (A - E) to relate the 5 CVMS to the 9 stages of Bjork hand-wrist analysis. RESULTS: In all age groups, the skeletal maturity stages of the hand and wrist bones and the cervical vertebrae of the girls were ahead of the boys. Cohen's kappa test revealed a low level of agreement between the two methods [Kappa (95% CI) = 0.312 (0.290 - 0.377)]; concordance was slightly higher in males (K = 0.33 for males versus 0.27 for females). Evaluation of concordance coefficients between the stages determined by the two methods indicated the highest concordance in 8- and 9-year-olds and the lowest in 12- and 14-year-olds. The level of agreement between the two methods was only acceptable in 8- and 9-year-olds of both genders and 10-year-old boys. The level of agreement between the two methods in other age groups was not acceptable. CONCLUSION: The level of agreement between the two methods was low; thus, they cannot be used alternatively to estimate patients' skeletal age or to predict the PGS. This may be due to the effect of different maturation levels (influenced by the environment, ethnicity, and gender) on the agreement between methods for skeletal age determination.