Comparison of two obturation techniques on adhesion and proliferation of human periodontal ligament fibroblasts / Comparación de la adhesión y proliferación de fibroblastos periodontales humanos en dos técnicas de obturación endodóntica

The present study aimed to compare the adhesion and proliferation of human periodontal ligament fibroblasts (hPDL) in transverse sections of the teeth sealed with two different obturation techniques, BioRoot RCS/hydraulic obturation (HO) and AH-Plus/continuous-wave condensation (CWC). The techniques were tested using an in vitro model to simulate the interaction between periodontal tissues and the materials. The root canals were instrumented and sterilized. A total of 15 samples were obturated with BioRoot RCS/HO and 15 samples with AH-Plus/CWC. Then, roots were sectioned to obtain obturated teeth slices, and hPDL cells were seeded onto the root slices. The results were obtained at intervals of 4 and 24h for cell adhesion; and at 3,7,14, and 21 days for cell proliferation. Empty cell culture plates were use as controls. The cell adhesion was increased at 4 and 24h for both groups, with an increased response observed in the BioRoot RCS/HO group (p<0.05). The difference in cell proliferation was also found between experimental groups. After 14 days of culture, BioRoot RCS/HO group showed an increase response than control and AH-Plus/CWC groups (p<0.05), and after 21 days both groups behaved better than control group, with an increased response observed in the BioRoot RCS/HO group. This study demonstrated that both root canal sealers allow the attach and growth of periodontal ligament fibroblasts, with an increased biological response in the BioRoot RCS/HO group.

El presente estudio se enfocó en comparar la adhesión y proliferación de fibroblastos de ligamento periodontal humano (hPDL) en secciones transversales de raíces previamente obturadas con dos técnicas de obturación diferentes: obturación hidráulica empleando cono único de gutapercha y BioRoot RCS como sellador (HO), y obturación de condensación de onda continua y AH-Plus como sellador (CWC). Los selladores se usaron en un modelo in vitro que simula la interacción entre los tejidos periodontales y los materiales de obturación. Los conductos radiculares fueron instrumentados, esterilizados y obturados. La muestra se compuso de un total de 15 raíces con la técnica BioRoot RCS/HO y 15 raíces con la técnica AH-Plus/CWC. Las células de hPDL fueron sembradas en condiciones estándar de cultivo sobre las raíces seccionadas. Los resultados fueron obtenidos a intervalos de 4 y 24h para adhesión celular, y a los 3,5,7,14 y 21 días de cultivo para proliferación celular. La adhesión celular a las 4 y 24 horas mostró ser diferente para ambas técnicas en comparación con el grupo control, siendo más importante en el grupo BioRoot RCS/HO. La diferencia en la proliferación entre grupos se observó a los 14 días de cultivo, únicamente para el grupo BioRoot RCS/HO; Sin embargo para el día 21 ambas técnicas mostraron mayor proliferación celular que el grupo control, con mejor respuesta para el grupo BioRoot RCS/HO. Este estudio ha demostrado que ambos selladores de conductos permiten la adhesión y crecimiento de fibroblastos de ligamento periodontal, siendo el grupo BioRoot RCS/HO el que mostró mayor biocompatibilidad.

Viability of periodontal ligament cells in selected transport media for avulsed teeth: an in vitro study

To compare the viability of periodontal ligament (PDL) cells stored in Hanks Balanced Salt Solution (HBSS) with those in readily available transport media over a variable period of time. Methods: Periodontal ligament cells harvested from premolars freshly extracted for orthodontic reasons were cultured for exponential growth. The cells were exposed to egg white, evaporated milk, water and Hanks Balanced Salt Solution (HBSS) at room temperature. Their viability was evaluated after 30 minutes, 1 hour and 3 hours with the tetrazolium salt-based colorimetric assay (MTT assay). Statistical analysis was done using the IBM® SPSS version 23.0 software. Comparison between the Mean Optical Densities (MODs) of the cells stored in HBSS and other media at each time interval was done using the independent t test. Repeated measure ANOVA and Tukey post-hoc test were also carried out to compare the MOD of cells within each medium over time. The level of significance was set at p <0.05. Result: The PDL cells stored in egg white had higher MODs than those in HBSS at 30 minutes and 1 hour. Conversely, the MODs of the cells stored in milk and water were lower than those in HBSS at all the studied points. There was a significant difference in the viability of the cells stored in HBSS and water at all the time points (p<0.05). Conclusion: For up to an hour, egg white was found to perform better than HBSS in supporting the viability of PDL cell

Channa Striata and Hyperbaric Oxygen Therapy Combination for Pressure Areas in Orthodontic Treatment

ABSTRACT Objective: To study the effect of using a combination of Channa Striata gel and hyperbaric oxygen therapy on pressure areas during orthodontic treatment. Material and Methods: The study was conducted using the ARRIVE Essential 10 guidelines. In this study, 35 3-4 months male guinea pigs (Cavia Cobaya) weighing 300-400 grams were used and divided into 5 groups (n=7). Decalcification was performed to dissolve the dental calcium and jawbone to cut the tissue properly. The decalcification was performed for 30 days. Then preparations were made with HE (Hematoxylin Eosin), observed using a microscope, and counted the number of osteoclasts and macrophages on a light microscope with 400 times magnification. The results of the preparations were analyzed using the SPSS program. Results: The Kruskal-Wallis test of macrophage cells and the ANOVA test of osteoclast cells showed significant results between all groups (p<0.05). Conclusion: The effect of hyperbaric oxygen therapy 2,4 ATA administered on days 8-14 and Channa Striata extract gel administered on days 3-14 can increase the number of macrophages in the periodontal ligament and osteoclasts in the alveolar bone in the pressure area during orthodontic tooth movement.

The peri-implant ligament: a scoping review

The peri-implant ligament is formed from the interface of bone tissue, through the anchoring of proteins and the surface of the dental implant. In this sense, it is relevant to understand the extent to which this ligament is structured and biomimics the periodontal ligament functions. Aim: The goal of this scoping review is to present and analyze the peri-implant ligament composition and compare the extent to which this ligament is structured and biomimics the periodontal ligament functions. Methods: This scoping review was performed according to the Joanna Briggs Institute methodology for scoping reviews and following the Preferred Reporting Items for Systematic Reviews and Meta-analyses extension for scoping review. Two independent researchers searched Pubmed, Cochrane, Embase, Virtual Health Library, Scielo, Scopus, Web of Science, Brazilian Bibliography of Dentistry, Latin American and Caribbean Literature in Health Sciences, Digital Library of Theses and Dissertations from the University of São Paulo and Portal Capes. Studies published in English, Portuguese and Spanish, over the last 21 years (2000-2021). Results: A total of 330 titles were identified and after applying inclusion and exclusion factors, 27 studies were included in this review. All proteins were identified regarding their tissue function and classified into 6 major protein groups. After that this new protein ligament was compared with the periodontal ligament regarding its function and composition. The main proteins associated with osseointegration, and thus, with the peri-implant ligament are recognized as belonging to the periodontal ligament. Conclusion: This scoping review results suggest evidence of the composition and function of the periimplant ligament. However, variations may still exist due to the existence of several modulants of the osseointegration process

Células madre en odontología: nuevas perspectivas / Stem cells in dentistry: new perspectives

En el campo de la odontología, prevalecen actualmente alternativas terapéuticas con una filosofía conservadora. Sin embargo, con el advenimiento de los tratamientos con células madre (CM), se amplían las posibilidades terapéuticas, que buscan la combinación y el equilibrio entre la intervención tradicional y las posibilidades de reposición de estructuras anatómicas dañadas, a través de la regeneración de tejidos utilizando células madre o sus derivados (AU)

In the dentistry field, therapeutic alternatives with a conservative philosophy currently prevail. However, with the advent of stem cell (SC) treatments, therapeutic possibilities are expanding, seeking a combination and balance between traditional intervention and the pos- sibility of replacing damaged anatomical structures through tissue regeneration, using stem cells or their derivatives (AU)

Viabilidade de fibroblastos do ligamento periodontal estimulados com sobrenadante necrótico de células de polpa dental in vitro / Viability of periodontal ligament fibroblasts stimulated with necrotic supernatant from dental pulp cells in vitro

As células necróticas são capazes de induzir a instalação de processos inflamatórios importantes, mesmo em ambientes estéreis, e o papel de remanescentes necróticos pulpares ainda não é conhecido na periodontite apical. O objetivo deste estudo foi investigar a capacidade de modulação in vitro de fibroblastos do ligamento periodontal (FLP) pelo sobrenadante necrótico (SN) de polpa dental. Cultura de fibroblastos de polpa dental (FPD) (n=1) e de ligamento periodontal (n=1) foram obtidas a partir do biobanco. Os FPDs passaram por ciclos de congelamento/descongelamento e o SN submetido a diluições 1/2, 1/10 e 1/20. Os FLPs foram estimulados com o SN de FPD. Para avaliar se o SN interfere na viabilidade celular do FLP foi realizado o ensaio de alamarBlue nos períodos de 24h, 48h e 72h. Nos grupos 24h, 48h e 72h não se observou diferença estatisticamente significativa. Porém, foi notado que no grupo de 72h o tratamento com SN resultou em uma maior ativação celular nas concentrações 1/10 e 1/20 em relação ao controle (p<00,5). O ensaio de MTT avaliou o efeito do SN na viabilidade celular do FLP. No grupo de 24h o SN resultou em uma ativação celular expressiva nas concentrações 1/2, 1/10 e 1/20 em relação ao controle. Nos grupos 48h e 72h não houve diferença estatisticamente significativa (p< 0,05). Os FLPs foram tratados com SN de FPD para avaliar os efeitos da produção de citocinas. A produção de OPG se apresentou maior no grupo com diluição 1/2 em comparação com as demais (p<0,05). A IL-6 apresentou-se aumentada no grupo com menor diluição e reduzida nos grupos com SN mais diluído (p<0,05). A expressão de CCL2 demonstrou-se menor nos grupos com diluição 1/10 e 1/20 (p<0,05). Diante desses resultados, observou-se que o sobrenadante de fibroblastos de polpa dental foi capaz de ativar células de ligamento periodontal in vitro modulando a produção de OPG, IL-6 e CCL2 de forma diluição-dependente.

Cytotoxicity of Reparative Endodontic Cements on Human Periodontal Ligament Stem Cells

ABSTRACT Objective To compare the cytotoxicity of commercial reparative endodontic cements on human periodontal ligament stem cells (hPDLSCs). Material and Methods The culture of hPDLSCs was established. Cell density was set at 2 × 104 cells/well in 96-well plates. Extracts of Biodentine, Bio-C Repair, Cimmo HD, MTA Repair HP and White MTA were prepared. Then, the extracts were diluted (pure, 1:4 and 1:16) and inserted into cell-seeded wells for 24, 48, and 72 h to assess cell viability through MTT assay. hPDLSCs incubated with culture medium alone served as a negative control group. Data were analyzed by Two-Way ANOVA and Tukey's test (α=0.05). Results At 24 h, pure extract of MTA Repair HP and Biodentine 1:16 presented higher cell viability compared to control. Lower cell viability was found for pure extract of Cimmo HD, MTA Repair HP 1:4 and 1:16, and White MTA 1:16. At 48 h, pure extract of Bio-C Repair and MTA Repair HP presented higher cell viability compared to control. At 72 h, only the pure extract of MTA Repair HP led to higher cell proliferation compared to control. Conclusion Biodentine, Bio-C Repair and MTA Repair HP were able to induce hPDLSCs proliferation. Cimmo HD and White MTA were found to be mostly cytotoxic in hPDLSCs.

Tratamento cirúrgico conservador de cisto radicular de grande extensão: relato de caso com acompanhamento / Conservative surgical treatment of large extension radicular cyst: case report with follow-up

Introdução: os cistos radiculares são as lesões císticas mais comuns nos maxilares. Eles surgem dos Restos Epiteliais de Malassez, presos no ligamento periodontal e podem ser ativados por um processo inflamatório na região pulpar. Geralmente são descobertos em exames radiográficos de rotina, apresentando-se como uma imagem radiolúcida, bem delimitada, envolvendo o periápice de um ou mais dentes. Objetivo: apresentar o tratamento de um extenso cisto radicular, em região de maxila, com acompanhamento de 18 meses. Relato do caso: Paciente do sexo feminino, 49 anos, foi encaminhada para avaliação e tratamento na Clínica Odontológica da Faculdade Sete Lagoas (FACSETE), apresentando lesão extensa em região maxilar anterior direita ao exame radiográfico. Ao exame clínico, observou-se leve assimetria facial e ausência de sintomas dolorosos. Tomografia computadorizada, punção aspirativa e biópsia incisional foram utilizadas para se chegar ao diagnóstico compatível com cisto radicular. Optou-se por uma técnica conservadora, em que foi realizada a descompressão da lesão. Após 05 meses de tratamento, um novo procedimento cirúrgico foi realizado para enuclear o restante da patologia. Conclusão: a descompressão, com utilização de cânula, é um tratamento auxiliar fácil, conservador, eficaz e reduz a morbidade causada por diferentes cistos odontogênicos(AU)

Introduction: root cysts are the most common cystic lesions in the jaw. They arise from the Epithelial Remains of Malassez, trapped in the periodontal ligament and can be activated by an inflammatory process in the pulp region. They are usually discovered in routine radiographic examinations, presenting as a well-defined radiolucent image involving the periapex of one or more teeth. Objective: to present the treatment of an extensive root cyst, in the maxillary region, with a follow-up of 18 months. Case report: A 49-year-old female patient was referred for evaluation and treatment at the Dental Clinic of Faculdade Sete Lagoas (FACSETE), with an extensive lesion in the right anterior maxillary region on radiographic examination. On clinical examination, mild facial asymmetry and absence of painful symptoms were observed. Computed tomography, aspiration puncture and incisional biopsy were used to reach a diagnosis compatible with radicular cyst. We opted for a conservative technique, in which the lesion was decompressed. After 05 months of treatment, a new surgical procedure was performed to enucleate the rest of the pathology. Conclusion: decompression, using a cannula, is an easy, conservative, effective auxiliary treatment and reduces the morbidity caused by different odontogenic cysts.

Cytotoxicity and cytokine production by calcium silicate-based materials on periodontal ligament stem cells

Abstract This study investigated the effect of three commercial calcium silicate-based materials (CSBM) on cytotoxicity and pro-and anti-inflammatory cytokines production in cultured human periodontal ligament stem cells (hPDLSCs). Culture of hPDLSCs was established and characterized. Extracts of Bio-C Sealer (Angelus, Londrina, PR, Brazil), MTA Fillapex (Angelus, Londrina, PR, Brazil) and PBS Cimmo HP (Cimmo Soluções em Saúde, Pouso Alegre, MG, Brazil) were prepared by placing cement specimens (5 x 3 mm) in culture medium. Then, the extracts were serially two-fold diluted (1, 1:2, 1:4, 1:8, 1:16) and inserted into the cell-seeded wells for 24, 48 and 72 h for MTT assays. TNF-α and IL-10 cytokines were quantified by ELISA at 24h-cell supernatants. Data were analyzed by ANOVA and Tukey's test (α = 0.05). All CSBM exhibited some cytotoxicity that varied according to extract concentration and time of evaluation. MTA Fillapex presented the highest cytotoxic effects with significant reduction of metabolic activity/cell viability when compared to Bio-C Sealer and Cimmo HP®. TNF-α was significantly upregulated by the three tested cements (p < 0.05) while only MTA Fillapex significantly upregulated IL-10 in comparison to control. Taken collectively, the results showed that PBS Cimmo HP®, Bio-C Sealer and MTA Fillapex present mild and transient cytotoxicity and slightly induced TNF-α production. MTA Fillapex upregulated IL-10 release by hPDLSCs.

Resumo Este estudo investigou o efeito de três materiais comerciais à base de silicato de cálcio (CSBM) na citotoxicidade e na produção de citocinas pró e antiinflamatórias em células-tronco do ligamento periodontal humano (hPDLSCs). Cultura de hPDLSCs foi estabelecida e caracterizada. Extratos de Bio-C Sealer (Angelus, Londrina, PR, Brasil), MTA Fillapex (Angelus, Londrina, PR, Brasil) e PBS Cimmo HP® (Cimmo Soluções em Saúde, Pouso Alegre, MG, Brasil) foram preparados com a colocação de espécimes dos cimentos (5 x 3 mm) em meio de cultura. Em seguida, os extratos foram diluídos (1, 1: 2, 1: 4, 1: 8, 1:16) e inseridos nos poços semeados de células para ensaio de citotoxicidade por meio de MTT por 24, 48 e 72 h. As citocinas TNF-α e IL-10 foram quantificadas por ELISA em sobrenadantes de células de 24 h. Os dados foram analisados por ANOVA e teste de Tukey (α = 0,05). Todos os CSBM exibiram alguma citotoxicidade que variou de acordo com a concentração do extrato e o tempo de avaliação. O MTA Fillapex apresentou os maiores efeitos citotóxicos com redução significativa da atividade metabólica / viabilidade celular quando comparado ao Bio-C Sealer e Cimmo HP®. O TNF-α foi regulado positivamente pelos três cimentos testados (p <0,05), enquanto apenas o MTA Fillapex regulou positivamente a liberação de IL-10 em comparação com o controle. Tomados em conjunto, os resultados mostraram que PBS Cimmo HP®, Bio-C Sealer e MTA Fillapex apresentam citotoxicidade leve e transitória e induziram a produção de TNF-α. O MTA Fillapex regulou positivamente a liberação de IL-10 por hPDLSCs.

Recombinant human insulin-like growth factor-1 promotes osteoclast formation and accelerates orthodontic tooth movement in rats

Abstract Background: IGF-1 may be an important factor in bone remodeling, but its mechanism of action on osteoclasts during orthodontic tooth movement is complex and unclear. Methodology: The closed-coil spring was placed between the left maxillary first molar and upper incisors with a force of 50 g to establish an orthodontic movement model. Eighty SD rats were randomized to receive phosphate buffer saline or 400 ng rhIGF-1 in the lateral buccal mucosa of the left maxillary first molar every two days. Tissue sections were stained for tartrate-resistant acidic phosphatase (TRAP), the number of TRAP-positive cells was estimated and tooth movement measured. Results: The rhIGF-1 group exhibited evidential bone resorption and lacuna appeared on the alveolar bone compared to the control group. Moreover, the number of osteoclasts in compression side of the periodontal ligament in the rhIGF-1 group peaked at day 4 (11.37±0.95 compared to 5.28±0.47 in the control group) after the orthodontic force was applied and was significantly higher than that of the control group (p<0.01). Furthermore, the distance of tooth movement in the rhIGF-1 group was significantly larger than that of the control group from day 4 to day 14 (p<0.01), suggesting that rhIGF-1 accelerated orthodontic tooth movement. Conclusion: Our study has showed that rhIGF-1 could stimulate the formation of osteoclasts in the periodontal ligament, and accelerate bone remodeling and orthodontic tooth movement.

Comparison of tooth movement and biological response resulting from different force magnitudes combined with osteoperforation in rabbits

Abstract Objective To compare tooth movement rate and histological responses with three different force magnitude designs under osteoperforation in rabbit models. Methodology 48 rabbits were divided into three groups: Group A, Group B, and Group C, with traction force of 50 g, 100 g, 150 g, respectively. Osteoperforation was performed at the mesial of the right mandibular first premolar, the left side was not affected. One mini-screw was inserted into bones between two central incisors. Coil springs were fixed to the first premolars and the mini-screw. Tooth movement distance was calculated, and immunohistochemical staining of PCNA, OCN, VEGF, and TGF-β1 was analyzed. Results The tooth movement distance on the surgical side was larger than the control side in all groups (P<0.01). No significant intergroup difference was observed for the surgical side in tooth movement distance among the three groups (P>0.05). For the control side, tooth movement distance in Group A was significantly smaller than Groups B and C (P<0.001); no significant difference in tooth movement distance between Group B and Group C was observed (P>0.05). On the tension area of the moving premolar, labeling of PCNA, OCN, VEGF and TGF-β1 were confirmed in alveolar bone and periodontal ligament in all groups. PCNA, OCN, VEGF and TGF-β1 on the surgical side was larger than the control side in all groups (P<0.001). Conclusion Osteoperforation could accelerate orthodontic tooth movement rate in rabbits. Fast osteoperforation-assisted tooth movement in rabbits was achieve with light 50 g traction.

Diagnosis of alveolodental ankylosis in unerupted canines: one of the answers to why the canine does not come

ABSTRACT Introduction: Teeth frequently fail to erupt and situations arise that prevent the canines from reaching the occlusal plane. Objective: Discourse about the three situations in which the canine does not reach the occlusal plane, and remains unerupted; and at the same time, point how to make a safe diagnosis of alveolodental ankylosis - one of the three causes -, based on tomography. Conclusions: Ankylosis occurs in impacted teeth by atrophy of the periodontal ligament, including the epithelial rests of Malassez. The tomographic signs of alveolodental ankylosis in unerupted canines are the interruption of hypodense periodontal space, discontinuity of the lamina dura and its continuity with the root surface, which gradually loses its regular shape.

RESUMO Introdução: Muitas vezes, a erupção falha, e ocorrem situações que impedem que os caninos cheguem até o plano oclusal. Objetivos: Discorrer sobre quais as três situações nas quais o canino não chega até o plano oclusal, permanecendo não irrompido e, ao mesmo tempo, destacar como se diagnosticar com segurança uma dessas três causas, a anquilose alveolodentária, a partir da tomografia. Conclusões: A anquilose em dentes não irrompidos ocorre pela atrofia do ligamento periodontal, incluindo os Restos Epiteliais de Malassez. Os sinais tomográficos de uma anquilose alveolodentária em caninos não irrompidos são a interrupção do espaço periodontal hipodenso, a descontinuidade da lâmina dura e a sua continuidade com a superfície radicular, que, gradativamente, perde sua regularidade.

Los restos de Malassez, vida y destino en los tejidos odontogénicos. Una revisión de la literatura / The remains of Malassez life and fate in the odontogenic tissues. A review of the kiterature

Los restos epiteliales de Malassez son células que se encuentran alrededor de las raíces de las piezas dentarias y forman parte de los tejidos del ligamento periodontal, donde se disponen en forma de red. Las funciones que desempeñan no son muy específicas; sin embargo, se demostró su participación en el mantenimiento del espacio del ligamento periodontal, la regeneración del tejido periodontal, la regeneración del cemento, entre otros. Además, tienen capacidad de diferenciarse en otros linajes celulares, lo que demuestra su capacidad como células madres, y el papel más conocido que tienen es su participación y proliferación en la formación de diversas patologías, como quistes y tumores odontogénicos. (AU)

Epithelial rests of Malassez are cells that are arranged in a network located around the roots of the teeth forming part of the periodontal ligament tissues. The functions that these cells perform are not very specific, however, they have shown to participate in the maintenance of the periodontal ligament space, regeneration of periodontal tissue, including cement regeneration among others. In addition, they are able to differentiate into other cell lineages, thereby demonstrating their capacity as stem cells. These cells are best known for their role in the participation and proliferation of the formation of different pathologies, such as cysts and odontogenic tumors. (AU)

Experimental acute sepsis reduced number of osteocalcin immunolabeled cells in periodontal ligament

Abstract The aim of this study was to evaluate the effect of acute sepsis in the periodontal ligament, alveolar and furcation bone in absence of periodontitis induction through histological and immunohistochemical analyses. A septic rat model was established by cecal ligation and puncture (CLP). Twelve rats were randomly divided into CLP (n=6) and Sham (n=6) groups. The animals were euthanized at 24 h and hemimandibles were submitted to histomorfometric (bone matrix, collagenous fibers, fibroblasts, osteocytes, inflammatory cells, and blood vessels) and immunohistochemical (BMP-2/4, RANKL and osteocalcin) evaluation in alveolar bone, furcation bone and periodontal ligament. Our results demonstrated that histomorphometric parameters were similar in alveolar bone, furcation bone and periodontal ligament of Sham and CLP rats. Regarding to immunohistochemical analyses, the number of BMP-2/4 and RANKL immunolabeled cells was also similar in both groups. Furthermore, it was detected a reduction in the osteocalcin immunolabeled cells in periodontal ligaments of CLP compared to Sham rats (p=0.0014). In conclusion, the acute sepsis induction resulted in reduced number of osteocalcin labelled cells in periodontal ligament region. Moreover, no significant histological differences were observed in the periodontium of rats under acute sepsis. Considering the role of osteocalcin in bone remodeling, the study contributes to revealing the importance of careful periodontal evaluation in the presence of sepsis.

Resumo O objetivo deste estudo foi avaliar o efeito da sepse aguda no ligamento periodontal, osso alveolar e osso da furca por meio de análise histológica e imunohistoquímica. O modelo de sepse em ratos foi estabelecido pelo procedimento de ligação e perfuração do ceco (CLP). Doze ratos foram divididos de forma randomizada em ratos sépticos (n=6) e controle - grupo Sham (n=6). Os animais foram eutanasiados após 24 horas e suas hemimandíbulas foram submetidas aos procedimentos histotécnicos para análise histomorfométricos (matriz óssea, fibras colágenas, fibroblastos, osteócitos, células inflamatórias e vasos sanguíneos) e imunohistoquímicos (BMP-2/4, RANKL e osteocalcina) no osso alveolar, osso de furca e ligamento periodontal. Nossos resultados demonstraram que os parâmetros histomorfométricos foram similares no osso alveolar, osso de furca e ligamento periodontal dos animais do grupo sepse e do grupo Sham. Em relação à análise por imunohistoquímica, o número de células imunomarcadas para BMP-2/4 e RANKL também foi similar em ambos os grupos. Entretanto, houve redução (p=0.0014) no número de células imunomarcadas para osteocalcina no ligamento periodontal de ratos sépticos em relação ao grupo Sham. Como conclusão, o estabelecimento de sepse aguda resultou em um número reduzido de células imunomarcadas para osteocalcina na região do ligamento periodontal (p=0,0014). Além disso, não foram observadas diferenças histológicas significativas no periodonto de ratos na presença de sepse aguda. Considerando o papel da osteocalcina na remodelação óssea, este estudo contribui para revelar a importância da avaliação periodontal cuidadosa na presença de sepse.

Reabsorções radiculares externas inflamatórias em dentes permanentes reimplantados: determinantes clínicos e epigenéticos / Reabsorções radiculares externas inflamatórias em dentes permanentes reimplantados: determinantes demográficos, clínicos e genético / Evaluation of Clinical Determinants and DNA Methylation profile in Inflammatory External Root Resorption in Replanted Permanent Teeth

A reabsorção radicular externa inflamatória (RREI) é um processo patológico definido como a perda progressiva de tecido mineralizado radicular, dentina e cemento, resultante da combinação entre a lesão às camadas protetoras da superfície externa da raiz e a presença de microrganismos no interior do sistema de canais radiculares. Estudos clínicos demonstraram o papel da idade e de fatores relacionados ao manejo e tratamento do dente avulsionado na etiopatogenia e evolução das RREI após reimplantes. Entretanto, não existem informações sobre a interação destes fatores, bem como poucos estudos avaliaram a influência do perfil genético e imunológico do paciente no padrão de cicatrização após reimplantes dentários. O presente estudo objetivou (1) avaliar a interação de fatores prognósticos para o desenvolvimento da RREI após o reimplante de dentes permanentes, bem como (2) investigar o papel da epigenética nos processos imunomediados das RREI pós-traumáticas. Para estudo dos determinantes clínicos e suas interações, o universo da pesquisa envolveu 427 pacientes (idade média de 12,6 anos) portadores de 581 dentes permanentes reimplantados, com rizogênese completa no momento do trauma, tratados na Clínica de Traumatismos dentários da Faculdade de Odontologia da Universidade Federal de Minas Gerais entre 1994 e 2018. Dados relativos à idade do paciente no momento do trauma, grau de rizogênese, condições de armazenamento e período extra alveolar do dente avulsionado, uso de antibioticoterapia sistêmica, tempo decorrido entre o reimplante e o início da terapia endodôntica radical (TER) e a duração do período de imobilização foram coletados dos prontuários dos pacientes. Tomadas radiográficas realizadas na consulta de início do TER foram utilizadas para diagnóstico da atividade de reabsorção. Sinais radiográficos de RREI foram encontrados em 80,7% da amostra (469 dentes). Os resultados demonstraram que a idade do paciente no momento do trauma e o tempo decorrido até o início do TER representaram importantes fatores prognósticos para a ocorrência de RREI. Além disso foi observada uma interação quantitativa entre estas duas variáveis uma vez que o aumento na idade do paciente atenuou significativamente o efeito do tempo até o início da terapia endodôntica. Este resultado inédito evidencia a maior vulnerabilidade do paciente mais jovem e enfatiza a importância de se considerar estas duas covariáveis conjuntamente durante a tomada de decisão clínica. Para o estudo epigenético, o perfil de metilação do DNA de 22 genes envolvidos na resposta imune foi avaliado em um pool de 08 amostras de fragmentos radiculares de dentes reimplantados portadores de RREI, indicados para exodontia. O grupo controle consistiu em um pool de 06 amostras de tecido ósseo saudável coletado durante a extração cirúrgica de dentes impactados. Os padrões de metilação do DNA dos 22 genes foram quantificados utilizando EpiTect Methyl II Signature Human Cytokine Production PCR Array. Os resultados do estudo da epigenética revelou que o pool de amostras com RREI apresentou nível mais alto de metilação do DNA na região promotora da FOXP3, em comparação com o pool de osso normal (65,95% e 23,43%, respectivamente). Esta é a primeira evidência de uma possível participação de eventos epigenéticos na modulação da RREI e especula-se se o padrão hipermetilado da FOXP3 poderia estar relacionado à presença da infecção endodôntica.

Inflammatory external root resorption (IERR) is a pathological process defined as the progressive loss of root mineralized tissue, dentin and cement, resulting from both: damage to the protective layers in the root external surface and the presence of endodontic infection inside the root canal. Clinical studies have demonstrated the role of age and factors related to the management and treatment of avulsed teeth in the etiopathogenesis and progression of RREI. However, there is no information on the interaction of these factors and few studies have evaluated the influence of the patient's genetic and immunological profile on the healing pattern after dental replantation. The present study aimed to (1) evaluate the interaction of prognostic factors for the development of RREI after replantation of permanent teeth, as well as (2) to investigate the role of epigenetics in the immunomediated processes of posttraumatic RREI. To study the clinical determinants and their interactions, the sample comprised 427 patients (mean age 12.6 years) with 581 replanted mature permanent teeth treated at the Dental Trauma Clinic of the Faculty of Dentistry from the Federal University of Minas Gerais between 1994 and 2018. Patients' records were evaluated to collect data such as patient's age at the time of the trauma, storage conditions and extra alveolar period of the avulsed tooth, systemic antibiotic therapy prescription, time elapsed between reimplantation and onset of endodontic therapy (TER) and splinting timing. The presence and index of IERR was assessed radiographically at the visit of pulpectomy. Radiographic signs of IEER were found in 80.7% of the sample (469 teeth) and were absent in 19.7% of cases (112 teeth). The results showed that the patient's age at the time of the trauma and the time that elapsed until the beginning of TER represented important prognostic factors for the occurrence of RREI. In addition, a quantitative interaction was observed between these two variables since the increase in the patient's age significantly attenuated the effect of time until the beginning of endodontic therapy. This is an original result that highlights the greater vulnerability of the younger patients and emphasizes the importance of considering these two covariates together during clinical decision-making. For the epigenetic study, the DNA methylation profile of 22 genes involved in the immune response was evaluated in a pool of 08 samples of root fragments of replanted teeth with RREI, referred to extraction. The control group consisted of a pool of 06 samples of healthy bone tissue collected during surgical extraction of impacted teeth. The DNA methylation pattern was quantified using EpiTect Methyl II Signature Human Cytokine Production PCR Array. The results of the epigenetics study revealed that the sample pool with RREI showed a higher level of DNA methylation in the FOXP3 promoter region, compared to the normal bone pool (65.95% and 23.43%, respectively). This is the first evidence of a possible participation of epigenetic events in the modulation of RREI and it is speculated whether the hypermethylated pattern of FOXP3 could be related to the presence of endodontic infection.

Effect of a novel bioceramic root canal sealer on the angiogenesis-enhancing potential of assorted human odontogenic stem cells compared with principal tricalcium silicate-based cements

Abstract Objective: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). Methodology: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. Results: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). Conclusion: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.

Pain Perception and Rate of Canine Retraction Through Self-Ligating Brackets and Conventional Elastomeric Ligation System: A Split Mouth Study

Abstract Objective: To evaluate the rate of tooth movement and the pain perception via self-ligating (SL) and conventional elastomeric ligation brackets (CB) system. Material and Methods: This study has been conducted at the Orthodontic Department of Baqai Dental College, Baqai Medical University. The sample size of this study comprised 40 patients, falling between the age of 12-30 years without any sex discrimination. Shapiro-Wilk was used to check the distribution of data. Non-parametric Mann Whitney U test was applied to evaluate the pain associated with SL and CB brackets system. To analysis the canine retraction Wilcoxon test was applied for the comparison of CB and SL brackets system. For all statistical analyses, the p-value of <0.05 was considered significant. Results: Pain level associated with retraction via CB and SL shows significant differences. However, the rate of canine retraction via CB and SL shows no significant differences at stages T0-T1 and T1-T2. However, stage T2-T3 shows a significant difference. Conclusion: As pain during orthodontic treatment is mostly associated with the level of compression of the periodontal ligament, it may be hypothesized that lower frictional forces generate less compression of the periodontal ligament and blood vessels, and so alter the type of pain experienced.

Estudo do processo de reparo do reimplante de dentes de ratos mantidos em albumina / Study of the reimplant repair process of teeth mainted on albumin

Quando o reimplante imediato não é possível, os meios de conservação têm papel fundamental para a manutenção da integridade do ligamento periodontal do dente avulsionado, para que um tratamento adequado seja alcançado. A clara do ovo apresentou resultados favoráveis para esse fim e a albumina constitui seu principal componente. Assim o propósito desse estudo foi analisar se a albumina pasteurizada em pó possui o mesmo potencial que a sua forma natural, como meio de conservação. Para isso foram utilizados 40 ratos divididos em 4 grupos de 10 animais. Após a extração do incisivo superior direito, os dentes do grupo IM foram reimplantados em seus respectivos alvéolos após 5 minutos (controle positivo grupo AP os dentes serão imersos em 40 ml de clara de ); No ovo pasteurizada em pó, reconstituída em água (10 gramas de pó em 30ml de água); No grupo CO dentes foram imersos em 40 ml de clara de ovo in natura; No grupo SE - os dentes serão mantidos em meio seco (controle negativo ). Com exceção do grupo IM, os demais dentes foram mantidos em seus respectivos meios por 60 minutos em temperatura ambiente para serem então reimplantados em seus respectivos alvéolos. Os animais receberam em dose única, antibiótico e analgésico. Na análise histológica da inserção epitelial e ligamento periodontal, foram consideradas a ocorrência de inflamação e a localização. Na superfície radicular foi analisada a ocorrência de reabsorção inflamatória, reabsorção por substituição, total de reabsorção, áreas reparadas por ligamento periodontal, anquilose. Os dados foram submetidos à análise estatística a um nível de significância de 5%. Os resultados demonstraram que a intensidade da inflamação aguda no tecido conjuntivo da inserção epitelial, foi maior no grupo SE em relação ao grupo IM. O grupo SE foi o mais comprometido que os demais grupos em relação à reabsorção total, reabsorção inflamatória e reabsorção por substituição. O grupo SE apresentou a menor extensão de ligamento periodontal que os demais grupos. Para o ligamento periodontal reinserido, o grupo SE apresentou menor extensão do que os grupo IM e AP. Pode-se concluir que a albumina em pó apresentou resultado histológico semelhante ao da clara de ovo in natura e do reimplante imediato, constituindo uma opção como meio de conservação de dentes avulsionados(AU)

When immediate replantation is not possible, preservatives play a fundamental role in maintaining the integrity of the periodontal ligament of the avulsed tooth so that proper treatment is achieved. Egg white showed favorable results for this purpose and a lbumin is its main component. Thus the purpose of this study was to analyze whether pasteurized albumin powder has the same potential as its natural form as a preservative medium. Forty rats were divided into 4 groups of 10 animals. After extraction of the right upper incisor, the teeth of the IM group were reimplanted in their respective alveoli after 5 minutes ( group AP positive control); In the teeth will be immersed in 40 ml of pasteurized egg white powder, reconstituted in water (10 grams of powder i n 30ml of water); In group CO were immersed in 40 ml of fresh egg white; In the SE group -- the teeth the teeth will be kept in a dry environment ( negative control). With the exception of the IM group, the remaining teeth were kept in their respective mea ns for 60 minutes at room temperature and then reimplanted in their respective alveoli. The animals received a single dose, antibiotic and analgesic. Histological analysis of epithelial insertion and periodontal ligament considered the occurrence of inflam mation and location. The root surface was analyzed for inflammatory resorption, replacement resorption, total resorption, areas repaired by periodontal ligament, ankylosis. Data were subjected to statistical analysis at a significance level of 5%. The resu lts showed that the intensity of acute inflammation in the connective tissue of the epithelial insertion was higher in the SE group than in the IM group. The SE group was the most compromised than the other groups in terms of total resorption, inflammatory resorption and replacement resorption. The SE group had the smallest periodontal ligament extension than the other groups. For the reinserted periodontal ligament, the SE group presented smaller extension than the IM and AP groups. It can be concluded tha t albumin powder presented histological results similar to that of fresh egg white and immediate replantation, constituting an option as a means of preserving avulsed teeth(AU)

Influence of photobiomodulation on pain perception during initial orthodontic tooth movement / Influência do fotobiomodulação na percepção da dor durante o movimento ortodôntico inicial

Introduction: Laser in low intensity (LLI) has been used to reduce the discomfort and pain that is triggered by the forces applied during orthodontic treatment. Objective: To evaluate the effect of LLI application in the pain perception of periodontal ligament initial compression, during orthodontic tooth movement; and to compare the effect of this therapy between men and women. Material and method: The sample consisted of 30 volunteers, who needed orthodontic band placement on mandibular first molars. After insertion of the elastic separators, LLI was applied to the mesial and distal apical region (wavelength 808nm, energy 2J, time 20s and fluency of 8.32J/cm2) and at three points on the root region (wavelength 808nm, energy 1J, time 10s and fluency of 4.16J/cm2) of the first molar (irradiated side) and compared to the contralateral first molar (non-irradiated side), in three time intervals: 0hs, 24hs and 48hs. Pain perception was evaluated by the Visual Analog Scale (VAS), at 0hs, 24hs and 48hs after insertion, with significance of 5%. Result: The pain level was observed to be significantly lower (p<0.05) on the irradiated side, irrespective of gender and time. Women presented a significantly higher pain level (p<0.05) than men, irrespective of time and side. There were no significant differences between the time intervals (p>0.05). Conclusion: It was concluded that LLI reduced the perception of initial pain in patients in whom compression of the periodontal ligament was promoted by elastic separation, and that women had a greater perception of pain sensitivity in the time intervals studied.

Introdução: O tratamento com Laser em Baixa Intensidade (LBI) tem sido utilizado para reduzir o desconforto e a dor desencadeados pelas forças aplicadas durante o tratamento ortodôntico. Objetivo: Avaliar o feito da aplicação de LBI na percepção da dor na compressão inicial do ligamento periodontal durante o movimento dentário ortodôntico; e comparar o efeito desta terapia entre os sexos. Material e método: A amostra foi composta por 30 voluntários, que necessitavam de bandagem dos primeiros molares inferiores. Após a instalação dos elásticos separadores, aplicou-se o LBI infravermelho na região apical mesial e distal (comprimento de onda 808nm, energia 2J, tempo 20s e fluência de 8,32J/cm2) e em três ponto na região radicular (comprimento de onda 808nm, energia 1J, tempo 10s e fluência de 4,16J/cm2) no lado irradiado e comparou-se com o primeiro molar contralateral não irradiado (lado controle), em três tempos: 0hs, 24hs e 48hs. A percepção de dor foi avaliada pela interpretação da Escala Visual Analógica (EVA) em 0hs, 24hs e 48hs após a instalação, com nílvel de significância de 5%. Resultado: Observou-se que o nível de dor foi significativamente menor (p<0,05) no lado irradiado, independentemente do sexo e do tempo. O sexo feminino apresentou nível de dor significativamente maior (p<0,05) que o sexo masculino, independentemente do tempo e do lado. Não houve diferença significativa entre os tempos (p>0,05). Conclusão: Concluiu-se que o LBI diminui a percepção de dor inicial em pacientes onde se promoveu a compressão do ligamento periodontal por meio de separação elástica, e que o sexo feminino apresentou maior percepção da sensibilidade dolorosa nos tempos observados.

Transcriptome profile of highly osteoblastic/cementoblastic periodontal ligament cell clones

Abstract Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale. Objective Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage. Methodology To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes. Results The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including "anatomical structure development" and "cell adhesion." Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. Conclusions This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process "anatomical structure development," Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.