Characterization of cells in blood evoked from periapical tissues in immature teeth with pulp necrosis and their potential for autologous cell therapy in Regenerative Endodontics.

OBJECTIVE: The objectives of this study were to isolate, characterize progenitor cells from blood in the root canals of necrotic immature permanent teeth evoked from periapical tissues and evaluate the applicable potential of these isolated cells in Regenerative Endodontics. DESIGN: Ten necrotic immature permanent teeth from seven patients were included. Evoked bleeding from periapical tissues was induced after chemical instrumentation of the root canals. Cells were isolated from the canal blood and evaluated for cell surface marker expression, multilineage differentiation potential, proliferation ability, and target protein expression. Cell sheets formed from these cells were transferred into human root segments, and then transplanted into nude mice. Histological examination was performed after eight weeks. Data analysis was conducted using one-way ANOVA followed by Tukey's post-hoc comparison, considering p < 0.05 as statistically significant. RESULTS: The isolated cells exhibited characteristics typical of fibroblastic cells with colony-forming efficiency, and displayed Ki67 positivity and robust proliferation. Flow cytometry data demonstrated that at passage 3, these cells were positive for CD73, CD90, CD105, CD146, and negative for CD34 and CD45. Vimentin expression indicated a mesenchymal origin. Under differentiation media specific differentiation media, the cells demonstrated osteogenic, adipogenic, and chondrogenic differentiation potential. Subcutaneous root canals with cell sheets of isolated cells in nude mice showed the formation of pulp-like tissues. CONCLUSIONS: This study confirmed the presence of progenitor cells in root canals following evoked bleeding from periapical tissues of necrotic immature teeth. Isolated cells exhibited similar immunophenotype and regenerative potential with dental mesenchymal stromal cells in regenerative endodontic therapy.

Efficacy of imaging techniques for the diagnosis of apical periodontitis: A systematic review.

BACKGROUND: Apical periodontitis (AP) is a chronic inflammatory response of microbial aetiology. Pathological changes associated with AP may not be visible on radiographic images and may linger without causing any symptoms. Clinicians rely mostly on clinical examination and imaging techniques to establish a diagnosis. OBJECTIVES: The aim of this review was to answer the following question using the PICO format: In the adult human permanent dentition (P), what is the efficacy of diagnostic imaging of the periapical tissues (I) using histopathology as a reference standard (C) in the diagnosis of apical periodontitis, in terms of diagnostic accuracy (O). METHODS: MEDLINE, EMBASE, Scopus and Cochrane Library were searched for English articles published through October 2021. At least two independent reviewers evaluated the study design, imaging modality used, histopathological assessment, outcome measures, results and conclusions for each article. The risk of bias was assessed using the Quality Assessment Tool for Diagnostic Accuracy Studies-2. RESULTS: The initial search strategy identified 544 articles. Seven articles were included for analysis in the final review, all of which involved tissue samples obtained from cadavers. No clinical studies were identified that met the eligibility criteria. A consistently low sensitivity score and negative predictive value were reported for periapical radiography, especially in comparison to CBCT, which scored highly. Both modalities achieved high scores for specificity and positive predictive value. Diagnostic accuracy of CBCT was lower for root-filled teeth in comparison to non-root-filled teeth. DISCUSSION: Assessment of the periapical tissues using periapical radiographs was shown to have a low to moderate agreement with the histopathological assessment. CBCT was reported to be more accurate than PR and demonstrated a good agreement with histopathology, especially for non-root-filled teeth. CONCLUSIONS: This review identified a need for greater standardization in methodology and reporting, and as the findings are based on cadaver studies, their clinical relevance must be interpreted with caution. REGISTRATION: PROSPERO (CRD42021272147).

The Potential Immunomodulatory Roles of Semaphorin 4D in Human Periapical Lesions.

INTRODUCTION: Semaphorin 4D (SEMA4D) is an important immunoregulator in the development of inflammatory diseases. Currently, the role of SEMA4D in human apical periodontitis remains unclear. This study aims to investigate the expression of SEMA4D and its potential immunomodulatory roles in apical periodontitis. METHODS: A total of 31 periapical tissues and 6 healthy gingival tissues were used in this experiment. Hematoxylin-eosin staining, immunohistochemical staining, and multiplex immunofluorescence staining were performed for histologic examination and immunochemical analysis. For data processing, the number of SEMA4D+, CD4+, CD8+, and CD20+ cells was analyzed by QuPath. In addition, the colocalization of SEMA4D with CD4, CD8, and CD20 was detected. RESULTS: Radicular cysts (RCs) (n = 18) and periapical granulomas (PGs) (n = 13) were identified by histologic evaluation. The number of SEMA4D+ cells in PGs was significantly greater than that in RCs (P < .05). T-cell and B-cell infiltration did not differ significantly between RCs and PGs. An increased number of CD20+ cells was observed in both types of apical periodontitis compared to CD8+ cells and CD4+ cells. Additionally, the presence of SEMA4D/CD4 and SEMA4D/CD20 double-positive cells was also markedly higher in PGs than in RCs. CONCLUSION: The expression of SEMA4D and related immune cells showed different characteristics between RCs and PGs. The disparate expression patterns indicated the possible different pathologic states of the 2 types of periapical lesions. This study provides a new perspective on the description of the comprehensive microenvironment of periapical lesions.

Unusual Lesion Caused by Food Displacement Into the Oral Mucosa, Mimicking a Sinus Tract.

This article reports an unusual case of foreign body lesion on the oral mucosa. A 61-year-old man, with no previous traumatic episode, presented with a small swelling resembling a sinus tract on the mucosa in the anterior mandible, adjacent to the left central and lateral incisors. Because of a discrete drainage of serous/purulent exudate, the clinician initially suspected endodontic involvement; however, the adjacent teeth were caries-free, responded positively to pulp tests, and the periapical tissues were radiographically normal. Exploration of the lesion opening under an operating microscope revealed a small seed associated with a dark filamentous structure; both were removed and processed for histologic examination. The seed was from a fig, and was germinating under the mucosa conditions. Bacterial colonization of the vegetable structures and polymorphonuclear leukocyte accumulations were histologically observed. The mucosa healed completely and uneventfully over the following weeks.

Inflammatory Response Mechanisms of the Dentine-Pulp Complex and the Periapical Tissues.

The macroscopic and microscopic anatomy of the oral cavity is complex and unique in the human body. Soft-tissue structures are in close interaction with mineralized bone, but also dentine, cementum and enamel of our teeth. These are exposed to intense mechanical and chemical stress as well as to dense microbiologic colonization. Teeth are susceptible to damage, most commonly to caries, where microorganisms from the oral cavity degrade the mineralized tissues of enamel and dentine and invade the soft connective tissue at the core, the dental pulp. However, the pulp is well-equipped to sense and fend off bacteria and their products and mounts various and intricate defense mechanisms. The front rank is formed by a layer of odontoblasts, which line the pulp chamber towards the dentine. These highly specialized cells not only form mineralized tissue but exert important functions as barrier cells. They recognize pathogens early in the process, secrete antibacterial compounds and neutralize bacterial toxins, initiate the immune response and alert other key players of the host defense. As bacteria get closer to the pulp, additional cell types of the pulp, including fibroblasts, stem and immune cells, but also vascular and neuronal networks, contribute with a variety of distinct defense mechanisms, and inflammatory response mechanisms are critical for tissue homeostasis. Still, without therapeutic intervention, a deep carious lesion may lead to tissue necrosis, which allows bacteria to populate the root canal system and invade the periradicular bone via the apical foramen at the root tip. The periodontal tissues and alveolar bone react to the insult with an inflammatory response, most commonly by the formation of an apical granuloma. Healing can occur after pathogen removal, which is achieved by disinfection and obturation of the pulp space by root canal treatment. This review highlights the various mechanisms of pathogen recognition and defense of dental pulp cells and periradicular tissues, explains the different cell types involved in the immune response and discusses the mechanisms of healing and repair, pointing out the close links between inflammation and regeneration as well as between inflammation and potential malignant transformation.

Immunohistochemical expression of PCNA, STRO-1 and CD 44 in the healing of experimentally induced periapical lesions in rats.

OBJECTIVE: The aim of the study was to determine the expression of cell proliferating marker, anti-proliferating cell nuclear antigen (anti-PCNA) and mesenchymal stem cell (MSC) markers (anti-STRO-1 and anti-CD44) in periapical periodontitis and their role in the healing of periapical lesion in periapical periodontitis. MATERIALS AND METHODS: Ninety Sprague-Dawley male rats (100 g) were divided into 3 groups: Experimental group I (EG I: n = 30), experimental group II (EG II: n=30) and control group (CG: n = 30). Periapical lesions were experimentally developed by leaving the dental pulp of maxillary first molars mesial root open to oral environment for 4 weeks. Conventional root canal treatment was performed in EG II. Maxillary first molars along with alveolar bone were resected and fixed. The processed samples were stained with routine hematoxylin and eosin (H&E), and evaluated immunohistochemically using antibodies against anti-PCNA, anti-STRO-1, and anti-CD44 polyclonal antibodies. Data were analyzed using Chi-square test and a p-value of <0.05 was considered significant. RESULTS: Immunostaining of anti-PCNA showed 30%, 70% and 53.3% positive staining in CG, EG I, and EG II, respectively (p<0.001). Moreover, the CD44 staining was 20% in CG in contrast to 63.6% in EG I and 43.3 in EG II. STRO-1 staining in CG was 10%, 50% in the EG I and 36.6% in the EG II (p<0.001). CONCLUSIONS: Periapical inflammatory tissues expressed significant proliferative cell marker PCNA and mesenchymal stem cell markers STRO-1, and CD44. These findings further reaffirm the promising role of mesenchymal stem cells in the healing of periapical periodontitis.

El tratamiento con implantes en pacientes con diabetes. Un estudio comparativo a 7 años / Treatment with zygomatic implants in patients with advanced atrophy of maxilla

INTRODUCCIÓN: La implantología oral puede constituir una modalidad terapéutica en el tratamiento prostodóncico de los pacientes con compromiso médico. El estudio muestra la evaluación del tratamiento con implantes en pacientes con diabetes comparados con pacientes sin diabetes. MÉTODOS: 48 pacientes edéntulos totales (24 pacientes diabéticos y 24 no diabéticos) fueron tratados con 96 implantes con superficie arenada y grabada Galimplant ® en la mandíbula para su rehabilitación prostodóncica con sobredentaduras mandibulares. 2 implantes fueron insertados en cada paciente. Los implantes fueron cargados funcionalmente tras un periodo de tiempo de 6 semanas con retenedores de fricción. Los hallazgos clínicos (implantológicos y prostodóncicos) se han seguido durante 7 años. RESULTADOS: Los resultados indican una supervivencia de los implantes del 95,8%. Durante el periodo de seguimeinto clínico se perdieron 4 implantes (dos implantes en cada grupo). La pérdida de hueso marginal media fué de 0,7 mm en ambos grupos. El 100% de los pacientes fueron tratados mediante una sobredentadura implantorretenida con anclajes de fricción. En 14 pacientes se realizaron cambios en los componentes plásticos de los ataches. El seguimiento clínico medio fue de 82,5 meses (60-102 meses). CONCLUSIONES: Los resultados del presente estudio indican que el tratamiento con implantes dentales en pacientes diabéticos es una terapéutica exitosa sin diferencias entre los pacientes diabéticos y no diabéticos

INTRODUCTION: Implant dentistry can to constitute a therapeutic modality in the prosthodontic treatment of medically compromised patients. This study reports the evaluation of treatment with implants in patients with diabetes compared with patients without diabetes. METHODS: 48 edentulous patients (24 diabetic patients and 24 non-diabetic patients) were treated with 96 Galimplant ® sand-blasted and acid-etched surface implants for prosthodontic rehabilitation with mandibular overdentures. Two implants were inserted in each patient. Implants were loaded after a healing free-loading period between 6 weeks with locator attaches. Clinical findings (implant and prosthodontics) were followed during at 7 years. RESULTS: Clinical results indicate a survival rate of implants of 95.8% in both groups. Four implant was lost during the follow-up period (two implants in each group). Media marginal bone loss was 0.7 mm in both groups. 100% of patients were treated with overdentures retained with 2 implants with locator attaches. Changes in plastic components of attaches were reported in 14 patients. The media follow-up was of 82.5 months (60-102 months). CONCLUSIONS: Clinical results of this study indicate that treatment with dental implants in diabetic patients is a successful implant treatment without differences with nondiabetic patients

Resolvin D2 Induces Resolution of Periapical Inflammation and Promotes Healing of Periapical Lesions in Rat Periapical Periodontitis.

Periapical periodontitis results from pulpal infection leading to pulpal necrosis and resorption of periapical bone. The current treatment is root canal therapy, which attempts to eliminate infection and necrotic tissue. But, in some cases periapical inflammation doesn't resolve even after treatment. Resolvins belongs to a large family of specialized pro-resolving lipid mediators that actively resolves inflammation signaling via specific receptors. Resolvin D2 (RvD2), a metabolite of docosahexaenoic acid (DHA), was tested as an intracanal medicament in rats in vivo. Mechanism was evaluated in rat primary dental pulp cells (DPCs) in vitro. The results demonstrate that RvD2 reduces inflammatory cell infiltrate, periapical lesion size, and fosters pulp like tissue regeneration and healing of periapical lesion. RvD2 enhanced expression of its receptor, GPR18, dentin matrix acidic phosphoprotein 1 (DMP1) and mineralization in vivo and in vitro. Moreover, RvD2 induces phosphorylation of Stat3 transcription factor in dental pulp cells. We conclude that intracanal treatment with RvD2 resolves inflammation and promoting calcification around root apex and healing of periapical bone lesions. The data suggest that RvD2 induces active resolution of inflammation with pulp-like tissue regeneration after root canal infection and thus maybe suitable for treating periapical lesions.

Development of criteria for investigation of periapical tissue from root-filled teeth.

OBJECTIVE: To develop and assess a set of criteria to grade inflammation including relative area of inflammation in periapical lesions in endodontically treated teeth. MATERIAL AND METHODS: A set of criteria was developed, encompassing data on: Lymphocytes, denoting chronic inflammation, were graded 0 (occasional) to 4 (heavy/dense inflammation). Polymorphonuclear cells, denoting acute inflammation, were graded 0 (none) to 2 (many). The third parameter, area of inflammation, that is, the relative area of the specimen that was inflamed, was graded 0 (none) to 4 (76-100%). The criteria were tested on 199 consecutive biopsies from 180 patients (aged 31-75 years). Information about symptoms was retrieved from the referrals. Mann-Whitney's U-test was used to calculate possible differences in average values for the histopathological variables in the two groups of patients: symptomatic or asymptomatic. RESULTS: Using the criteria, varying grades of inflammation were seen in the biopsies. The majority showed few or no PMN cells. There was a correlation between symptoms and the extent of infiltration of lymphocytes and plasma cells (p = .001), PMN cells (p < .001) and the area of inflammation (p = .002): biopsies from the asymptomatic patients exhibited less pronounced and relatively smaller areas of inflammation. CONCLUSIONS: Using the criteria on a specific selection of root-filled teeth with persisting apical periodontitis, periapical inflammation was common, but varied in extent and severity. The inflammation was less pronounced and affected a relatively smaller area in asymptomatic teeth, although outliers in both directions were identified.

Immunohistochemical Analysis of Cyclooxygenase-2 and Tumor Necrosis Factor Alpha in Periapical Lesions.

INTRODUCTION: The purpose of this study was to evaluate the expression of cyclooxygenase-2 (COX-2) and tumor necrosis factor alpha (TNF-α) in periapical granuloma (PG) and radicular cyst (RC) samples and to correlate it with the type of lesion, the intensity of the inflammatory infiltrate, and the thickness of the epithelial lining. METHODS: A total of 51 cases of periapical lesions (25 PGs and 26 RCs) were subjected to morphologic analysis and immunohistochemical study. The anti-COX-2 and anti-TNF-α antibodies were applied using the immunoperoxidase technique. Data were analyzed by the Mann-Whitney test, Pearson chi-square test, Fisher exact test, and Spearman correlation. RESULTS: Analysis of the inflammatory infiltrate revealed that 80% of PGs exhibited a grade III infiltrate as opposed to a 19% rate in RCs (P < .001). Morphologic evaluation of the epithelial thickness of RCs revealed the presence of atrophic epithelium in 73% of cases. The majority of PGs had a score of 1 for COX-2 immunoexpression (n = 14, 54%) and a score of 2 for TNF-α expression (n = 16, 64%), whereas in cases of RCs a score of 1 was more prevalent for COX-2 and TNF-α expression (n = 17, 65%). Significant differences in the expression scores of COX-2 and TNF-α were detected in periapical lesions (P < .001). CONCLUSIONS: Based on these findings, we emphasize that RCs and PGs have a similar expression of inflammatory mediators (COX-2 and TNF-α) although the secretion of TNF-α by macrophages and of COX-2 by several cells was higher in PGs, indicating a greater inflammatory response in these lesions.

Randomized controlled clinical study of veneered zirconia abutments for single implant crowns: Clinical, histological, and microbiological outcomes.

OBJECTIVES: To analyze the effect of veneering of the submucosal part of zirconia abutments and the type of retention (cemented vs screw-retained) on clinical, microbiological, and histological outcomes of single-tooth implant crowns. MATERIAL AND METHODS: A total of 44 patients with a single missing tooth to be replaced by an implant in the anterior region participated in the study. Implants were randomly assigned to receive zirconia-based CAD/CAM reconstructions using either one of four treatment modalities: cement-retained with submucosal veneering (CR-P), cement-retained without submucosal veneering (CR-W), screw-retained with submucosal veneering (SR-P), and screw-retained without submucosal veneering (SR-W). Clinical parameters were assessed at baseline (after crown insertion), at 6 and 12 months. Histological and microbiological analyses were performed at 6 months. Descriptive statistics and the Kruskal-Wallis test were applied. RESULTS: The clinical evaluation revealed, in general, stable peri-implant soft tissues with minimal differences for all measured parameters between the four groups, except for bleeding on probing with the two cemented groups exhibiting higher values at 12 months (35.0% ± 26.5% for CR-W and 25.0% ± 38.8% for CR-P versus 13.1 ± 14.8 for SR-W and 13.0 ± 18.2 for SR-P). The descriptive and semi-quantitative histology showed a trend for a higher inflammatory reaction in the two cemented (a medium to high number of inflammatory cells) compared to the screw-retained groups (low number of inflammatory cells) at 6 months. The microbiological test demonstrated low bacterial counts and a similar distribution in between the groups except for two species (Tannerella forsythia and Peptostreptococcus micros) that were found in higher counts in the cemented groups at 6 months. CONCLUSION: Submucosal veneering of zirconia abutments did not negatively affect the health of the peri-implant tissues. The cemented groups, though, did show a clinical and histological trend to higher levels of inflammation.

Histology of NeoMTA Plus and Quick-Set2 in Contact with Pulp and Periradicular Tissues in a Canine Model.

INTRODUCTION: NeoMTA Plus (Avalon Biomed Inc, Bradenton, FL) is a tricalcium silicate material similar to the first mineral trioxide aggregate product, ProRoot MTA (Dentsply Sirona, York, PA), but with improvements such as decreased setting time, increased ion release, increased water sorption, and nonstaining radiopacifiers. Quick-Set2 (Avalon Biomed Inc) is a newly formulated calcium aluminosilicate material that has a faster setting time and increased acid resistance and is nonstaining. The purpose of this study was to compare the healing of pulpal and periapical tissues in dogs after exposure to NeoMTA Plus and Quick-Set2 after pulpotomy and root-end surgery procedures. METHODS: Seventy-two teeth (36 for each procedure) in 6 beagle dogs received pulpotomy or root-end surgery using either NeoMTA Plus or Quick-Set2. The dogs were sacrificed at 90 days, and the teeth and surrounding tissues were prepared for histologic evaluation. Sixty teeth were evaluated and scored histologically (29 with pulpotomies and 31 with root-end resections). Specimens were scored for inflammation, quality and thickness of dentin bridging, pulp tissue response, cementum and periodontal ligament formation, and apical bone healing. RESULTS: Both materials displayed favorable healing at 90 days. The only significant difference was the quality of dentin bridge formation in pulpotomies using NeoMTA Plus compared with Quick-Set2. CONCLUSIONS: Quick-Set2 and NeoMTA Plus had similar effects on inflammation, pulp response, periodontal ligament and cementum formation, and apical tissue healing in dogs. NeoMTA Plus had superior dentin bridge quality compared with Quick-Set2.

Tejido hepático ectópico intratorácico / Ectopic Intrathoracic Liver Tissue

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Large Bacterial Floc Causing an Independent Extraradicular Infection and Posttreatment Apical Periodontitis: A Case Report.

This article describes a case of large persistent posttreatment apical periodontitis associated with 2 maxillary incisors, which was successfully managed by periradicular surgery. Histobacteriologic analysis revealed that the lesion was a granuloma that contained in its body a very large actinomycoticlike colony surrounded by accumulations of polymorphonuclear leukocytes and showing no direct communication with the root canal systems from both teeth. One incisor had no evidence of persistent intraradicular infection, whereas the other exhibited some residual dentinal tubule infection in the apical canal, which may have not significantly contributed to persistent inflammation given the organization and agglomeration of inflammatory cells around the large extraradicular bacterial colony. Findings showed that the main cause of persistent disease was the extraradicular infection in the form of a large bacterial floc, apparently independent of an intraradicular infection and as such only solved by surgery.

The Expression of Interferon Regulatory Factor 8 in Human Periapical Lesions.

INTRODUCTION: Interferon regulatory factor 8 (IRF8) is a critical transcription factor in innate immune responses that regulates the development and function of myeloid cells. Human periapical lesions are caused by endodontic microbial infections. However, the presence of IRF8 in human periapical lesions remains elusive. This study aims to explore the expression of IRF8 in human periapical lesions and the possible association of IRF8 with macrophages, nuclear factor kappa B (NF-κB) signaling, and the autophagy process. METHODS: Thirty-nine human periapical tissues, including healthy control tissues (n = 15), radicular cysts (RCs, n = 11), and periapical granulomas (PG, n = 13), were examined. Tissues were fixed in paraformaldehyde and analyzed. The inflammatory infiltrates of lesions were evaluated by hematoxylin-eosin, and the expression of IRF8 was analyzed by immunohistochemistry. Double immunofluorescence assessment was performed to colocalize IRF8 with CD68, NF-κB p65, and LC3B. RESULTS: The expression of IRF8 was significantly higher in RCs and PGs than in the healthy control group, but no significant difference was found between RCs and PGs. There were significantly more IRF8-CD68 double-positive cells in RCs and PGs than in the healthy control group, but no significant difference was observed between RCs and PGs. Double-labeling analysis of IRF8 with NF-κB and LC3B indicated that IRF8 expression is associated with NF-κB signaling and the autophagy process during periapical lesions. CONCLUSIONS: IRF8 could be observed and might possibly be involved in macrophages in the development of periapical lesions.

Comparative Expression of CD34, Intercellular Adhesion Molecule-1, and Podoplanin and the Presence of Mast Cells in Periapical Granulomas, Cysts, and Residual Cysts.

INTRODUCTION: The aim of the present study was to compare the immunoexpression of CD34, intercellular adhesion molecule-1 (ICAM-1), and podoplanin and the presence of mast cells with clinical, demographic, radiologic, and histologic features from periapical granulomas, periapical cysts, and residual cysts. METHODS: Thirty-one lesions (5 granulomas, 15 periapical cysts, and 11 residual cysts) were selected. Histologic sections in silanized slides were used for the immunohistochemical reactions. The analysis of the images was performed by using an optical microscope, and data were analyzed with 5% significance (P < .05). RESULTS: Cysts presented atrophic and hyperplastic epithelium in 11 cases (35.5%) and 15 cases (48.8%), respectively (P > .05). The intensity of the inflammatory infiltrate was similar when comparing the 3 groups (P > .05). CD34 and podoplanin expression and the presence of mast cells were similar when comparing the 3 groups; ICAM-1 expression was more intense in granulomas than cysts (P < .05). There were no statistically significant differences associated with the expression of the evaluated markers according to the intensity of the inflammatory infiltrate. CONCLUSIONS: There were no differences in the expression of CD34 and podoplanin and in the presence of mast cells when the 3 groups were compared. ICAM-1 expression was more common in periapical granulomas.

Influence of apical enlargement on the repair of apical periodontitis in rats.

AIM: To evaluate the influence of different apical enlargement protocols on the radiographic and histological healing of apical periodontitis in rats. METHODOLOGY: Apical periodontitis was induced bilaterally in the mandibular right and left first molars of 24 Wistar rats by pulp exposure to the oral cavity for 3 weeks. A standard serial root canal preparation technique was performed in the molar of one side, whilst the opposite side was the control group. Rats were randomly divided into three experimental groups (n = 8), according to the diameter of apical enlargement during root canal preparation: K-files size 20 (EG1), size 25 (EG2) and size 30 (EG3). Each animal was its own positive control, because the opposite arch remained untreated. Root canals were filled with a standard technique. After 3 weeks, the animals were euthanized. The main outcome of apical periodontitis healing was evaluated radiographically (mm2 ) and histologically (ordinal scores of inflammation) using a HE staining technique. The measurement of effect was obtained between the three experimental groups by carrying out generalized estimating equations, with Poisson regression with robust variance, pairing each experimental group with its respective control group within animals, adjusted for the mean within animal differences, with α = 5%. RESULTS: The mean and standard deviations of radiographic apical periodontitis size (mm2 ) and intensity of histological inflammatory scores were, respectively: EG1 (0.44 ± 0.27; 2.25 ± 0.46), EG2 (0.33 ± 0.10; 2.50 ± 0.53) and EG3 (0.22 ± 0.08; 2.63 ± 0.74). After 3 weeks, a significantly more favourable radiographic repair was observed when larger apical enlargement was performed (EG3), compared to EG1 and EG2 (P = 0.001). All experimental groups were associated with a significant difference on the radiographic and histological healing of apical periodontitis compared with its respective control group. CONCLUSION: Under the experimental conditions of this study, a larger apical enlargement protocol favoured a more rapid radiographic repair of apical periodontitis in rats after a 3-week follow-up.

Evaluation of Microleakage by Gas Permeability and Marginal Adaptation of MTA and BiodentineTM Apical Plugs: In Vitro Study / Evaluación de la microfiltración por permeabilidad de gas y adaptación marginal de tapones apicales de MTA y BiodentineTM: Estudio in vitro

Abstract The endodontic treatment of teeth with incomplete development is always a complex task. Nowadays, biomaterials such as bioceramics offers promising clinical evidence that supports its use. However, the standardization of its use for apexification purpose still needs a deeper understanding of the materials' behavior. The aim of this investigation was to evaluate the marginal adaptability and microleakage by gas permeability of MTA and Biodentine™ apical plugs in an in vitro model. Materials and methods: Twenty- four single rooted human teeth were selected according to previously stablished inclusion criteria. All samples were prepared obtaining standard cylindrical internal canals with a diameter of 1.3 mm. Root canals were gently rinsed using 5.25% sodium hypochlorite and EDTA 17%. The apical 3mm and remaining coronal dental structure were sectioned to obtain 10mm roots. Roots were randomly assigned to 3 different groups as follows: GROUP A: MTA (n=10), GROUP B: Biodentine™ (n=10) and Group C: Control (positive n=1, negative n=3). MTA and Biodentine™ were prepared according to manufacturer's indications, and apical plugs of 4mm were passively placed in the correspondent teeth. All samples were stored in saline solution for 7 days at 37°C before evaluation. Samples were mounted in cylindrical sample-holders using epoxy resin. Microleakage was evaluated with an automatic permeability detector that calculates nitrogen diffusion between the material-root interphase. After microleakage evaluation, the samples were recovered and analyzed by scanning electron microscopy (SEM). Microleakage results were analyzed using Chi-square and adaptation was evaluated with a descriptive analysis. Results: None of the evaluated materials completely avoided the nitrogen microleakage (positive leakage of 10% and 20% of samples for MTA and Biodentine™ respectively); with no statistical significant difference between groups (p=0.527). All apical plugs showed good adaptation under SEM, at 30x, 200x, 1000x and 2500x; with microscopical structures similar to previous reports. Conclusions: Both bioceramics behave similar when used as apical barriers to avoid permeability, with acceptable marginal adaptation. Further in vivo studies are needed to validate these results.

Resumen El tratamiento endodóntico de dientes con desarrollo incompleto es siempre una tarea compleja. Hoy en día, biomateriales como las biocerámicas ofrecen una evidencia clínica prometedora que apoya su uso. Sin embargo, la estandarización de su uso para fines de apexificación todavía necesita una comprensión más profunda del comportamiento de los materiales. El objetivo de esta investigación fue evaluar la adaptabilidad marginal y microfiltración por permeabilidad de gas de los tapones apicales de MTA y Biodentine™ en un modelo in vitro. Materiales y métodos: Veiticuatro dientes humanos uniradiculares fueron seleccionados meticulosamente según criterios de inclusión previamente establecidos. Todas las muestras fueron preparadas con canales cilíndricos internos estandarizados de 1,3 mm de diámetro. Los conductos radiculares fueron gentilmente lavados con hipoclorito de sodio al 5,25% y EDTA al 17%. La estructura dental apical de 3 mm y la coronal restante se seccionó para obtener raíces de 10 mm de longitud. Las raíces se asignaron aleatoriamente a 3 grupos diferentes de la siguiente manera: GRUPO A: MTA (n = 10), GRUPO B: Biodentine™ (n = 10) y Grupo C: Control (n = 1 positivo, n = 3 negativos). El MTA y Biodentine™ se prepararon de acuerdo con las indicaciones de los fabricantes, y se colocaron pasivamente los tapones apicales de 4 mm en los dientes correspondientes. Todas las muestras se almacenaron en solución salina durante 7 días a 37ºC antes de la evaluación. Las muestras se montaron en porta-muestras cilíndricos utilizando resina epóxica. La microfiltración se evaluó con un detector de permeabilidad automática que calcula la difusión de nitrógeno entre la interfase material-raíz. Después de la evaluación de microfiltración, las muestras fueron recuperadas y analizadas por microscopía electrónica de barrido (SEM). Los resultados de microfiltración se analizaron utilizando una prueba estadística de Chi-cuadrado y la adaptación se evaluó con un análisis descriptivo. Resultados: Ninguno de los materiales evaluados evitó completamente la microfiltración de nitrógeno (fuga positiva de 10% y 20% de muestras para MTA y Biodentine™, respectivamente); sin diferencias estadísticamente significativas entre los grupos (p = 0,527). Todos los tapones apicales mostraron una buena adaptación bajo SEM, a 30x, 200x, 1000x y 2500x; con morfologías similares a las previamente reportadas. Conclusiones: ambas biocerámicas se comportan de forma similar cuando se usan como barreras apicales para evitar la permeabilidad de gas, con adaptación marginal aceptable. Se necesitan más estudios in vivo para validar estos resultados.

Periapical Cemento-osseous Dysplasia Is Rarely Diagnosed on Orthopantomograms of Patients with Neurofibromatosis Type 1 and Is Not a Gender-specific Feature of the Disease.

Several skeletal aberrations of the skull have been described for the tumor predisposition syndrome neurofibromatosis type 1 (NF1). Recently, periapical cemental/cemento-osseous dysplasia (COD) has been described in females affected with NF1. This reactive lesion of the hard tissues in tooth-bearing areas of the jaw has been proposed to represent a gender-specific radiological feature of NF1. The aim of this study was to investigate the prevalence of COD in patients with NF1. PATIENTS AND METHODS: The orthopantomograms (OPGs) of 179 patients with a confirmed diagnosis of NF1 were analyzed for COD. The results were compared to radiographic findings obtained in OPGs of age- and sex-matched controls. The NF1 patient group was further differentiated according to the evidence of facial plexiform neurofibroma. RESULTS: COD was a very rare finding in both groups. The extension of the diagnostic criteria including radiologically-healthy teeth and a widened periodontal gap in the periapical area only marginally increased the number of considered cases. Although there was a somewhat more common occurrence of such changes in the patient group compared to the control group and the number of affected women was greater than the number of men, none of these differences reached statistical significance. Furthermore, COD or widening of the periradicular periodontal space was not found to be associated with facial tumor type in NF1. CONCLUSION: The investigation revealed that COD is not a diagnostic feature of NF1. There is no clear association of the rare finding of COD with gender. These studies should be compared with patient groups of other ethnic backgrounds.

Immunohistochemical Analysis of Galectins-1, -3, and -7 in Periapical Granulomas, Radicular Cysts, and Residual Radicular Cysts.

INTRODUCTION: Galectins play important roles in immunoinflammatory responses, but their participation in the development of periapical lesions remains unclear. This study aimed to evaluate the expressions of galectins-1, -3, and -7 in periapical lesions, correlating them with the intensity of the inflammatory infiltrate and the pattern of the cystic epithelium. METHODS: Twenty periapical granulomas (PGs), 20 radicular cysts (RCs), and 20 residual radicular cysts (RRCs) were submitted to immunohistochemistry using anti-galectin-1, -3, and -7 antibodies. The percentage of immunopositive cells in epithelial and connective tissues was determined. RESULTS: In connective tissue, PGs exhibited higher cytoplasmic/membrane expression of galectins-1 and -7 than RCs and RRCs (P < .05). There was higher nuclear expression of galectin-1 in PGs compared with RCs and RRCs (P < .05). The expression of galectins-1 and -7 in connective tissue was higher in lesions with grade III inflammation (P < .05). No significant differences in galectin-3 immunoexpression were observed for any of the parameters evaluated (P > .05). In the epithelial component, a higher nuclear expression of galectin-7 was detected in RRCs (P < .05), and a higher cytoplasmic/membrane expression of this protein was found in cysts with hyperplastic epithelium (P < .05). Positive correlations were observed between the nuclear and cytoplasmic/membrane expression of galectin-1 in connective tissue (P < .05) as well as between the nuclear and cytoplasmic/membrane expression of galectin-7 in epithelial tissue of cysts (P < .05). CONCLUSIONS: Galectins-1 and -7 may play important roles in the pathogenesis of PGs, RCs, and RRCs. On the other hand, the present results suggest only a minor involvement of galectin-3 in the development of these lesions.