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.

Substance P/calcitonin gene-related peptide and their receptors expression in human periodontal ligament after root canal preparation with five different systems.

AIM: The purpose of this study was to quantify the effect of five different root canal preparation instruments on Substance P (SP), Calcitonin gene-related peptide (CGRP) and their receptors expression in healthy human periodontal ligament. METHODOLOGY: STROBE guidelines were used to design a study using 60 periodontal ligament samples obtained from healthy lower premolars where extraction was indicated for orthodontic reasons. Prior to extraction 40 of these premolars were equally divided into four groups and root canals were prepared using different systems: Mtwo, Reciproc Blue, HyFlex EDM and Plex-V. Ten premolars were prepared with hand files and served as a positive control group. The remaining 10 premolars where extracted without treatment and served as a negative control group. All periodontal ligament samples were processed to measure the expression of SP, CGRP and their receptors by radioimmunoassay. Kruskal-Wallis and Duncan tests were performed to determine statistically significant differences between the groups for each variable. RESULTS: Greater expression of all the peptides measured were found in the hand-file preparation group, followed by the Reciproc Blue, Mtwo, HyFlex EDM and Plex-V groups. The lower SP, CGRP and their receptors values were for the intact teeth control group. Kruskal-Wallis test showed statistically significant differences amongst groups (p < .001). Dunn post-hoc tests showed statistically significant differences in SP, CGRP and their receptors expression between the intact teeth and the hand-file and Reciproc Blue groups. Hand-file group showed significant differences with the other groups, except with Reciproc Blue, where no differences were observed in any of the peptides measured. Finally, no differences were observed between Plex-V and HyFlex in any of the peptides measured. CONCLUSIONS: Root canal preparation with hand files and Reciproc Blue generates the highest expression of SP, CGRP, NK1 and CGRP1R in human periodontal ligament, whilst Plex-V and HyFlex maintain the basal expression of neuropeptides and their receptors. Mtwo showed intermediate results between Reciproc Blue and HyFlex.

Allogeneic Bone Marrow Mesenchymal Stromal Cell Transplantation Induces Dentin Pulp Complex-like Formation in Immature Teeth with Pulp Necrosis and Apical Periodontitis.

INTRODUCTION: Dental pulp regeneration is challenging in endodontics. Cellular therapy is an alternative approach to induce dental pulp regeneration. Mesenchymal stromal cells (MSCs) have the capacity to induce dental pulp-like tissue formation. In this study, we evaluated the capacity of allogeneic bone marrow MSCs (BM-MSCs) to regenerate pulp following necrosis and apical periodontitis in children's permanent immature apex teeth. METHODS: Patients aged 8 to 12 years with pulp necrosis and apical periodontitis were evaluated. The study included 15 teeth (13 incisors and 2 molars) from 14 patients (8 boys and 6 girls). Radiographic evaluation showed periapical radiolucency and immature apex teeth. There was no response to cold or electric pulp testing. The root canal of each tooth was cleaned, shaped, and Ca(OH)2 used as an interappointment medication. Cryopreserved allogeneic BM-MSCs were thawed, expanded, incorporated into preclotted platelet-rich plasma, and implanted into the tooth's pulp cavity. They were sealed with bioceramic cement and composite. Sensibility, apical foramen, calcium deposits within the root canal, and resolution of periapical lesions were evaluated in each tooth over the following 12 months. RESULTS: Based on 9 variables established for dental pulp-like tissue regeneration, all MSC-treated teeth showed evidence of successful regeneration. Clinical and radiographic evaluation of the treated teeth showed periapical lesion healing, sensitivity to cold and electricity, decreased width of the apical foramen, and mineralization within the canal space. CONCLUSIONS: Transplantation of allogeneic MSCs induces the formation of dental pulp-like tissue in permanent immature apex teeth with pulp necrosis and apical periodontitis. Implant of MSCs constitutes a potential therapy in regenerative endodontics in pediatric dentistry. Future studies incorporating a larger sample size may confirm these results.

Effectiveness, efficiency, and apical extrusion of 2 rotaries and 2 reciprocating systems in removing filling material during endodontic retreatment. A systematic review.

Background: This systematic review investigated the effectiveness, efficiency and apical extrusion of the debris of two rotary and two reciprocating single-file systems used for the removal of filling material from straight root canals. Material and Methods: A literature search was performed in the Medline, ISI Web of Science, and Scopus databases for relevant articles matching the keyword search strategy. Effectiveness was determined with studies dealing with the ability of the instruments to remove filling material from root canals. Efficiency was assessed with studies dealing with the time needed to completely remove the root canal filling, and apical extrusion was determined with studies that measured the amount of filling material extruded through the apex. Results: From the 424 articles initially found, 406 were excluded for being non-relevant or not fulfilling the selection criteria. Another 9 articles were excluded after methodology evaluation. Finally, 9 studies were included in the systematic review. Conclusions: None of the reviewed systems is effective to completely remove the filling materials from straight root canals, and all systems appear to be equally time-efficient, although this variable shows different results. In terms of apical extrusion, the analyzed reciprocating systems extrude more material toward the periapical tissues than the continuous rotation systems. Key words:Systematic review, rotary files, reciprocating files, apical extrusion, endodontic retreatment.

The effect of orthodontic forces on calcitonin gene-related peptide (CGRP) expression in the human periodontal ligament and its relationship with the human dental pulp.

Background: The purpose of this study was to quantify the effect of moderate and severe orthodontic forces on Calcitonin gene-related peptide (CGRP) expression in the healthy human periodontal ligament (PDL) and its possible relationship with the human dental pulp. Material and Methods: Ninety human periodontal ligament samples were obtained from healthy premolars where extraction was indicated for orthodontic reasons. Prior to extraction, teeth were divided in 3 groups of 30 samples each: I) Untreated teeth control group; II) Moderate force group: A 56 g force was applied to the premolars for 24 hours; and III) Severe force group: A 224 g force was applied to the premolars for 7 days. All periodontal ligament samples were processed and CGRP was measured by radioimmunoassay. Results: Greater CGRP expression was found in the severe force group, followed by the moderate force group. The lower CGRP values were for the untreated teeth. Kruskal-Wallis test showed statistically significant differences between groups (p<0.001). LSD post hoc tests showed statistically significant differences in CGRP expression between the untreated teeth and the severe forces group (p<0.001). Differences between the moderate and severe force groups were statistically significant (p<0.001). There was no statistically significant differences between the untreated teeth and the moderate forces group (p<0.261). Conclusions: CGRP expression in human periodontal ligament increases when teeth are submitted to severe orthodontic forces. This elevated expression of CGRP, which is proportional to the applied force, may affect the way the dental pulp responds to different stimuli from the orthodontic forces. Key words:Calcitonin gene-related peptide, orthodontic force, human periodontal ligament, neurogenic inflammation.

In Vitro Comparison of MTA and BC RRM-Fast Set Putty as Retrograde Filling Materials.

OBJECTIVE: To compare, in vitro, the bioceramic materials (MTA and BC RRM-fast set putty) capacity to prevent microleakage of Enterococcus faecalis over time. METHODS: An experimental design was made with forty extracted human teeth, coronally cut, and prepared to be placed in a leakage system under sterile conditions. They were randomly divided into two experimental groups: thirty teeth (fifteen for each of retrograde filling material MTA and BC RRM-fast set putty) and a control group: ten teeth (five positive control, five negative control). The 3 mm root-ends were submerged in a brain-heart infusion broth with a red phenol indicator. The coronal access of each sample was inoculated with E. faecalis every seven days to maintain bacterial viability. The lower chamber was evaluated daily for 30 days to observe the turbidity of the culture medium and establish the presence and day of the filtration. Calculation of the colony-forming units (CFU) was performed for each leaked sample. Fisher's Exact Test was used to verify the association between the presence or absence of leakage of the samples by type of bioceramic material used and the Mann-Whitney U test to verify the existence of a difference between the average of CFU by type of bioceramic material used. The significance level used was α=0.05 and a 95% confidence level, as a decision rule for rejecting the null hypothesis. RESULTS: Of the total samples prepared for each group, leakage was found in 60.0% (9/15) of the MTA group and 40.0% (6/15) of the BC RRM-fast set putty group. All positive controls filtered on the first day of evaluation, while 20% (1/5) of the negative control leaked in the second week. There was no significant difference in leakege between the two groups, nor concerning the bacterial count (P=0.101) and the type of cement used (P=1.000). CONCLUSION: BC RRM-fast set putty was comparable to MTA in resisting bacterial microleakage during the observation time.

The hypoxia-dependent angiogenic process in dental pulp.

BACKGROUND: In this review, we analyzed the existing literature to elucidate how the hypoxia-dependent angiogenic processes work in dental pulp. Angiogenesis is an essential biological process in the maturation and homeostasis of teeth. It involves multiple sequential steps such as endothelial cell proliferation and migration, cell-to-cell contact, and tube formation. HIGHLIGHT: Clinical implications of understanding the process of angiogenesis include how the mineralization processes of dental pulp occur and how dental pulp maintains its homeostasis, preventing irreversible inflammation or necrosis. CONCLUSION: The angiogenesis process in dental pulp regulates adequate concentrations of oxygen required for mineralization in root development and defense mechanisms against chronic stimuli.

Dental Pulp Regeneration Induced by Allogenic Mesenchymal Stromal Cell Transplantation in a Mature Tooth: A Case Report.

INTRODUCTION: Cellular therapy constitutes a new therapeutic alternative in regenerative endodontics. In this case report, we evaluated the capacity of allogeneic mesenchymal stromal cells (MSCs) to induce dental pulp and apical bone regeneration in a tooth previously endodontically treated. METHODS: A healthy 55-year-old female patient consulting for swelling and a sinus tract associated with tooth #8 was referred for an endodontic evaluation. Previously, tooth #8 had undergone root canal treatment and apical resection and had no response to thermal or electric pulp testing. Radiographically, tooth #8 showed root canal treatment, a cut apex angle, and periapical radiolucency. The root canal was recleaned and shaped, and calcium hydroxide was used as an interappointment medication. Cryopreserved allogeneic bone marrow MSCs were thawed, expanded, incorporated into preclotted platelet-rich plasma, and implanted into the pulp cavity of tooth #8. The cervical part of the canal was sealed with bioceramic cement and a composite. RESULTS: After 14 months of MSC transplantation, tooth #8 showed sensitivity to cold and electric pulp tests. Radiographic and cone-beam computed tomographic imaging showed signs of increased periapical bone density, healing of the periapical lesion, and almost complete apical remodeling. CONCLUSIONS: This case report shows periodontal bone formation, apex remodeling, and dental pulp regeneration induced by allogeneic MSC transplantation in a mature nonvital tooth. Allogeneic MSCs may constitute a first-line therapy in regenerative endodontics.

Gene Expression of Vascular Endothelial Growth Factor A and its Receptors in Dental Pulp of Immature and Mature Teeth.

OBJECTIVE: Vascular endothelial growth factor A (VEGFA) and its receptors are essential proteins for the angiogenic activity of dental pulp. Angiogenesis fundamentally provides oxygen and nutrients to cells for root formation and defence mechanisms. The angiogenic potential of dental pulp should be understood and considered for the conservative and regenerative endodontics. The purpose of this research was to measure the VEGFA expression and its receptors such as vascular endothelial growth factor receptors 1, -2 (VEGFR1, VEGFR2) and Neuropilin 1 (NRP1) in human dental pulp from molars with immature and mature apexes. METHODS: VEGFA system mRNAs expressions were assessed in dental pulp obtained from freshly extracted human third molars divided into immature (n=8) and mature (n=8) apexes. RNAs were extracted from the samples. Each sample's cDNA was synthetized and the target genes VEGFA, VEGFR1, VEGFR2, NRP1 expression profiles obtained by RT2-PCR. Analysis was based on the Student's t-test comparing the replicate 2-ΔCt values for each gene. P values of <0.05 were considered significant. RESULTS: In teeth with mature apexes, VEGFA (P=0.0002), NRP1 (P=0.0001), VEGFR1 (P=0.0057) and VEGFR2 (P=0.018259) significantly increased statistically with respect to the immature apexes group. CONCLUSION: Within the limitation of the present investigation, it can be concluded that the angiogenic process seems to be a physiological process in the dental pulp due to the studied angiogenic growth factor are expressed in both immature and mature dental pulps. VEGFA and its receptors are expressed significantly higher in mature apex teeth than immature apex teeth.

Root canal morphology and variations in mandibular second molars: an in vivo cone-beam computed tomography analysis.

BACKGROUND: The purpose of this study was to determine the anatomical variations of the root canal system of mandibular second molars using cone-beam computed tomography (CBCT). METHODS: 190 mandibular second molars cone-beam computed tomography images were reviewed. The evaluation was performed by a radiologist with endodontic experience and two endodontists trained with CBCT technology. Tooth position, number of root and root canals, C-shaped root canal system configuration, presence of extra root (radix), and radicular grooves were assessed. Data was statistically analyzed using The Chi-square test (α = 0,05) to determine any significant difference between gender and the total number of root and root canals, and any significant difference between gender and root canal anatomical variation. RESULTS: Overall, 85.5% showed two separated roots, 12.1% a single root, 2.6% three roots or radix. 87.7% showed three root canals, 12.1% two root canals, 2.6% four root canals, and 1.6% a single root canal. 10% showed a single foramen, 75.3% two foramina, 13.6% three foramina and 1% showed four foramina.19.5% showed C-shaped anatomical variation, 51.4% in male patients, 48.6% in female patients. According to Fan classification: C1 13.6% in cervical third, C2 10% in the middle third, C3 17.3% in middle third, 15.5% in apical third, and C4 12.7% in the apical third. Root canals number in these samples were 5.4% a single canal, 21.6% two canals, 70.3% three canals, and 2.7% four canals. The root showed 46% with one foramen, 46% two foramina, and 8% three foramina. Radicular grooves 83.3% were found in the lingual area and 16.2% towards the buccal area. CONCLUSIONS: The most prevalent anatomic presentation of the evaluated sample was a mandibular second molars with two roots, three root canals, and two apical foramina. Their variation was C-shaped root canals and Radix Paramolaris.

Cone-beam computed tomography evaluation of C-shaped root and canal morphology of mandibular premolars.

BACKGROUND: Mandibular premolars are complicated teeth to endodontically treat due to the anatomical variations that can present. The purpose of this study was to determine the presence of C-shaped configurations in mandibular premolars by cone-beam computed tomography (CBCT). METHODS: 380 mandibular first premolars and 308 mandibular second premolars cone-beam computed tomographic images were obtained from 292 patients (175 female and 117 male). Tooth position, number of roots, root canals, C-shaped root canal system configuration, level of canal bifurcation and radicular grooves were evaluated by two endodontists trained in CBCT evaluation; previously calibrated between them, and a radiologist with endodontic experience. Data were statistically compared by The Chi-square test (α = 0.05) to examine any significant difference between gender and C-shaped root canal system and any significant difference between C-shaped configuration according to Fan criteria and gender. RESULTS: Overall 100% (n = 688) teeth examined, 19.2% (n = 132) had a C-shaped root canals system. 9.16% (n = 63) in male patients and 10.03% (n = 69) in female patients. The prevalence of C-shaped root canal system in mandibular first premolar was 83.33 and 16.66% in mandibular second premolars. According to Fan classification, the mandibular first premolars showed 3.63% as C1, 9.09% C2, 59.09% C3, 21.81% C4a, 1.8% C4b. Mandibular second premolars showed 13.63% as C1, 18.18% C2, 13.63% C3, 27.27% C4a, 9.09% C4b. Male patients showed 4.54% as C1, 3.78% C2, 22.72% C3, 11.36% C4a, 0.75% C4b, and 4.54% out of classification. Female patients showed 0.75% as C1, 6.81% C2, 30.03% C3, 9.84 C4a, 2.27% C4b, and 2.27% out of classification. The 53.36% canal bifurcation in mandibular first premolars and 50.09% in mandibular second premolars were in the middle third. No statistical differences were found between C-shaped root canal system and gender or C-shaped configuration according to Fan criteria and gender. P was < 0.05. CONCLUSIONS: Within the limitation of this study, the presence of C-shaped anatomical system is relatively low in mandibular second premolars than in mandibular first premolars.

Expression of substance P, calcitonin gene-related peptide and vascular endothelial growth factor in human dental pulp under different clinical stimuli.

BACKGROUND: The aim of this study was to measure the dental pulp inflammatory response through neuropeptides (SP and CGRP) as a response to occlusal trauma, orthodontic movements and a combination of both, as well as the angiogenic defense mechanism through VEGF expression, which could be the initial step to mineralized tissue formation. METHODS: Forty human dental pulp samples were collected from healthy first premolars with extraction indicated due to orthodontic reasons from a sample of 20 patients. Patients were divided into four groups with 10 premolars each (1 mandibular and 1 maxillary premolar from each patient): healthy pulp control group, occlusal trauma group, moderate orthodontic forces group; and occlusal trauma plus moderate orthodontic forces group. Stimuli were applied for 24 h before tooth extraction in all experimental groups. All samples were processed, and SP, CGRP, and VEGF were measured by radioimmunoassay. The Kruskal-Wallis test was performed to assess significant differences among groups and Mann-Whitney's U post hoc pairwise comparisons were also performed. RESULTS: The highest increase in SP, CGRP, and VEGF expressions was found in the occlusal trauma plus orthodontic forces group, followed by the moderate orthodontic forces, the occlusal trauma and the control groups, with statistically significant differences between all groups for each of the 3 peptides analyzed (Kruskal-Wallis p < 0.001). All possible pairwise post-hoc comparisons were also significant for each peptide analyzed (Mann-Whitney's U p < 0.001). CONCLUSION: SP, CGRP, and VEGF expressions significantly increase in human dental pulps when stimulated by occlusal trauma combined with moderate orthodontic forces, as compared with these two stimuli applied independently. Name of the registry: Importance of Neurogenic Inflammation in the Angiogenic Response of the Dental Pulp as a Defensive Response. TRIAL REGISTRATION NUMBER: NCT03804034. Date of registration: 01/15/2019 Retrospectively registered. URL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03804034?term=NCT03804034&draw=2&rank=1 .

Expression of early angiogenesis indicators in mature versus immature teeth.

BACKGROUND: Proper oxygen balance in the dental pulp is essential for cell metabolism. Angiogenesis in the pulp is a constant process during the life of the tooth. Hypoxia indicators in a tissue, such as HIF-1α, as well as vascular destabilization markers, such as ANG2 and its receptor TIE2, are necessary for angiogenesis. Therefore the purpose of this study is to evaluate the expression of HIF-1α, ANG1, ANG2 and TIE2 in dental pulp as early angiogenesis indicators in teeth with complete and incomplete root development. METHODS: Forty human dental pulps were obtained from freshly extracted third molars divided into two groups: incomplete (n = 20) and complete (n = 20) root development. Dental pulps were stored at - 80 °C, defrosted in an ice bath and re-frozen with their respective thaws to disintegrate the tissue. Three sonication cycles were performed until the tissues were homogenized, then thaw were centrifuged and the supernatant was collected for the detection of the markers to be studied. The samples were processed for the ELISA test using the ELISA-sandwich principle. Student t and Mann-Whitney U tests were performed to determine statistically significant differences between groups. RESULTS: In the complete root development, HIF-1α, ANG1, ANG2 and TIE2 expressions were significantly higher than their expression in the incomplete root development group. CONCLUSIONS: The angiogenic process seems to be a physiological process in the dental pulp. Angiogenic activity is higher in teeth with mature than immature apex teeth.