Avances en endodoncia regenerativa: reporte de caso con uso de la membrana amniótica humana / Advances in regenerative endodontics: report of a case using a human amniotic membrane

Los procedimientos endodónticos regenerativos (REPs) representan una evolución significativa en el campo de la endodoncia, buscando no sólo tratar la infección o lesión presente en el diente, sino tam-bién promover la regeneración de los tejidos denta-rios afectados. El presente caso clínico muestra un incisivo lateral superior izquierdo con apexogénesis incompleta y diagnóstico de absceso alveolar crónico reagudizado en una paciente de 22 años, en el que se aplicó un procedimiento de endodoncia regenerativa (REPs). La estrategia terapéutica elegida se basó en los principios de ingeniería tisular, incorporando la novedosa aplicación de la membrana amniótica hu-mana liofilizada esterilizada como andamio bioactivo intraconducto. Las evaluaciones clínicas, radiográ-ficas y tomográficas a corto, mediano y largo plazo revelaron el éxito de la terapia. La resolución exitosa mostró en los controles a la pieza dentaria asintomá-tica, con una notable remisión de la patología apical, aumento de la longitud radicular y disminución del calibre apical. Se ha podido destacar la eficacia de los REPs, con una exitosa aplicabilidad de la membra-na amniótica como andamio innovador (AU)

Regenerative endodontic procedures (REPs) represent a significant evolution in the field of endodontics, aiming not only to address the infection or injury within the tooth, but also to promote the regeneration of the affected dental tissues. In this clinical case, an upper left lateral incisor with incomplete apexogenesis and diagnosis of acute exacerbation of a chronic periapical lesion in a 22-year-old patient is presented. A regenerative endodontic procedure (REPs) was applied. The chosen therapeutic strategy was based on tissue engineering principles, incorporating the innovative use of sterilized lyophilized human amniotic membrane as an intraconduct bioactive scaffold. Clinical, radiographic, and tomographic assessments at short, medium, and long-term follow-up revealed the success of the therapy. Successful resolution demonstrated an asymptomatic tooth in the follow-up, with a notable resolution of apical pathology, increased root length, and decreased apical caliber. The effectiveness of REPs has been highlighted, demonstrating the successful applicability of amniotic membrane as an innovative scaffold (AU)

Determinación de parámetros estéticos gingivales y dentarios en dientes anterosuperiores / Determination of gingival and dental esthetic parameters in anterosuperior teeth

La odontología estética no es una disciplina especial o área de la odontología en sí misma, pero con consideraciones funcionales y biológicas, representa uno de los objetivos de las intervenciones de tratamiento dental, que abarca todas las áreas de especialidad, desde la odontología preventiva y restaurativa hasta la prostodoncia, ortodoncia, periodoncia, como, así como la cirugía oral y maxilofacial. El cénit gingival es un importante componente de la sonrisa y su estudio sigue siendo muy relevante en la odontoestética internacional. En el presente trabajo se determinaron distancias del cenit al eje longitudinal y se correlacionaron con otros parámetros gingivales, se establecieron diferencias en las alturas de las papilas interdentales y se correlacionaron los datos métricos de las piezas dentarias anteriores de la población estudiada y los datos conocidos con el fin de obtener datos estadísticos relevantes. Los datos mesurables fueron obtenidos de pacientes de ambos sexos, (18 ­ 25) años, con piezas dentarias del grupo anterior y superior a saber: incisivos centrales, incisivos laterales y caninos superiores, normalmente implantados, libres de lesiones o restaruración, ausencia de enfermedad gingivoperiodontal y sin tratamientos ortodoncicos. El trabajo observacional, descriptivo y tranversal arrojó resultados basados en la estadística preponderante. Provee dimensiones y proporciones de dientes maxilares que pueden adaptarse a pacientes individuales en relación con parámetros establecidos en la odontoestética. Estos datos pueden ser pautas útiles para el diagnóstico y la planificación del tratamiento (especialmente cirugía periodontal) en la dentición maxilar (AU)

A esthetic dentistry is not a special discipline or area of dentistry itself, but with functional and biological considerations, it represents one of the objectives of dental treatment interventions, which covers all areas of specialty, from preventive and restorative dentistry up to prosthodontics, orthodontics, periodontics, as well as oral and maxillofacial surgery. The gingival cenith is an important component of the smile and its study remains very relevant in international dentistry. In this work, distances from the cenith to the longitudinal axis were determined and correlated with other gingival parameters, differences in the heights of the interdental papillae were established and the metric data of the anterior teeth of the studied population were correlated and the known data with in order to obtain relevant statistical data. The measurable data were obtained from patients of both sexes, (18 - 25) years, with teeth of the anterior and superior group, namely: central incisors, lateral incisors and upper canines, normally implanted, free of lesions or restoration, absence of disease gingivoperiodontal and without orthodontic treatments. The observational, descriptive and transverse work produced results based on the preponderant statistics. It provides dimensions and proportions of maxillary teeth that can be adapted to individual patients in relation to parameters established in odontoesthetics. These data can be useful guidelines for diagnosis and treatment planning (especially periodontal surgery) in the maxillary dentition (AU)

Reconstrucción de la papila interdental con ácido hialurónico. Artículo de Revisión Corto / Interdental papilla reconstruction with hyaluronic acid. Mini review

Uno de los problemas estéticos odontológicos más frecuentes es la presencia de triángulos negros por la pérdida de la papila interdental. Existen varias opciones de tratamiento quirúrgico y no quirúrgico para corregir esta anomalía, entre las más novedosas y menos invasivas, encontramos la utilización de un gel inyectable de ácido hialurónico para la reconstrucción de la papila interdental. El objetivo del estudio fue realizar una recopilación bibliográfica sobre la efectividad del ácido hialurónico como tratamiento para la reconstrucción de la papila interdental.

One of the most frequent cosmetic dental problems is the presence of black triangles due to the loss of the interdental papilla. There are several surgical and non-surgical treatment options to correct this anomaly, among the most innovative and less invasive we found the use of an injectable gel of hyaluronic acid for the reconstruction of the interdental papilla. The objective of the study was to carry out a bibliographic compilation on the effectiveness of hyaluronic acid as a treatment for the reconstruction of the interdental papilla.

Berberine mediates root remodeling in an immature tooth with apical periodontitis by regulating stem cells from apical papilla differentiation / 国际口腔科学杂志·英文版

Once pulp necrosis or apical periodontitis occurs on immature teeth, the weak root and open root apex are challenging to clinicians. Berberine (BBR) is a potential medicine for bone disorders, therefore, we proposed to apply BBR in root canals to enhance root repair in immature teeth. An in vivo model of immature teeth with apical periodontitis was established in rats, and root canals were filled with BBR, calcium hydroxide or sterilized saline for 3 weeks. The shape of the roots was analyzed by micro-computed tomography and histological staining. In vitro, BBR was introduced into stem cells from apical papilla (SCAPs). Osteogenic differentiation of stem cells from apical papilla was investigated by alkaline phosphatase activity, mineralization ability, and gene expression of osteogenic makers. The signaling pathway, which regulated the osteogenesis of SCAPs was evaluated by quantitative real time PCR, Western blot analysis, and immunofluorescence. In rats treated with BBR, more tissue was formed, with longer roots, thicker root walls, and smaller apex diameters. In addition, we found that BBR promoted SCAPs osteogenesis in a time-dependent and concentration-dependent manner. BBR induced the expression of β-catenin and enhanced β-catenin entering into the nucleus, to up-regulate more runt-related nuclear factor 2 downstream. BBR enhanced root repair in immature teeth with apical periodontitis by activating the canonical Wnt/β-catenin pathway in SCAPs.

Mediated pathways of exosomes uptake by stem cells of apical papilla / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the uptake of exosomes by stem cells from apical papilla (SCAP), thus to provide experimental basis for mechanism of the exosomes endocytosis by SCAP.@*METHODS@#(1) Exosomes of dental pulp stem cells (DPSCs) were isolated by hypercentrifugation combined with ultrafiltration method. The exosomes were identified by transmission electron microscopy, nanoparticle tracking analysis and western blot. (2) PKH-26 membrane labeling technology was used to mark the DPSCs derived exosomes. The labeled exosomes were co-cultured with SCAP at 37 °C as positive control group, and co-cultured with SCAP at 4 °C as the low-temperature treatment group, while the negative control group was set up. (3) Using clathrin-mediated endocytosis inhibitor chlorpromazine (CPZ, 10 μmol /L) as CPZ group, caveolae-mediated endocytosis Genistein (200 μmol/L) as Genistein group, and macropinocytosis inhibitor LY294002 (50 μmol/L) as LY294002 group to treat the SCAP respectively. Solvent control group (DMSO group) was set. Immunofluorescence staining was used to detect the red fluorescence SCAP and flow cytometry was used to analyze the proportion of SCAP labeled with red fluorescence.@*RESULTS@#(1) The bilayer membrane and cup-shaped appearance of representative exosomes were observed. The peak of the size of DPSCs-derived exosomes was at 144 nm. The exosomes expressed exosomal marker proteins TSG101 and CD63, but not GAPDH which was the cellular internal control protein. (2) Immunofluorescence staining showed that after being co-cultured at 37 °C for 6 hours, red fluorescence could be detected in SCAP but it could not be detected after being co-cultured at 4 °C for 6 hours. After endocytosis inhibition, the red fluorescence in SCAP was reduced. Flow cytometry showed that the proportion of SCAP labeled with red fluorescence in positive group was 35.0%, in negative control group was 0.5%, and in solvent control group was 29.7%, in CPZ group, Genistein group and Genistein group were reduced to 13.7%, 16.6%, and 20.9%, respectively.@*CONCLUSION@#SCAP could uptake the DPSCs derived exosomes, and low temperature could inhibit this process. The exosomes uptake of SCAP was mediated by the clathrin endocytosis pathway, caveolae-mediated endocytosis and macropinocytosis pathway.

Remodelación de papila gingival interdental con ácido hialurónico. Una solución estética / Remodeling interdental gingival papilla with hyaluronic acid. An aesthetic solution

RESUMEN: El objetivo de este trabajo es evaluar el efecto de remodelación y relleno mediante aplicación de ácido hialurónico en papilas gingivales de sector estético con defectos de triángulo negro en un paciente tratado periodontalmente. Se inyectaron 0.15 ml de gel de ácido hialurónico dividido en 3 sesiones. Clínicamente se observó aumento de volumen y aumento de dimensiones verticales y horizontales en las papilas, mejorando así los parámetros estéticos. Como conclusión la aplicación de ácido hialurónico es efectiva en la remodelación y relleno en pérdidas de papila clase I de Nordland y Tarnow.

ABSTRACT: The objective of this work is to evaluate the effect of remodeling and filling through the application of hyaluronic acid in gingival papilla in the aesthetic sector with black triangle defects, in a periodontally- treated patient. In 3 portioned sessions, 0.15 ml of hyaluronic acid gel was injected. Clinically, an increase in volume was observed, and vertical and horizontal dimensions in the papilla increased too, thus improving aesthetic parameters. In conclusion, the application of hyaluronic acid is effective in the remodeling and filling in losses of class I papilla (Nordland and Tarnow classification).

Una Nueva Alternativa para la Creación de Papilas en Implantes de Carga inmediata: Reporte de Caso / A New Alternative for the Creation of Papillae in Immediate Loading Implants: Case Report

RESUMEN: El objetivo de este artículo es evaluar el éxito clínico de una nueva alternativa para crear papilas en implantes de carga inmediata, que se realizó en una paciente de sexo femenino de 26 años de edad. Se inició el tratamiento con la confección de una guía quirúrgica, previo a la manipulación de los tejidos para crear las papilas peri-implantarias. En la cirugía se realizan 2 incisiones semilunares, unidas por una incisión perpendicular, las que al suturarlas sobre ellas mismas permitió crear las papilas; se instala un implante con buena estabilidad primaria y se toma la impresión para la rehabilitación. A las 24 horas se instala una corona atornillada, mostrando la presencia de las papilas de forma inmediata. Se realiza controles hasta los 20 meses de función, evidenciando buena estabilidad de las papilas mesial y distal sin presentar ningún tipo de complicación.

ABSTRACT: The aim of this paper is to evaluate the clinical success of a new alternative to create papillae in immediate load implants, which was performed in a 26-yearold female patient. The treatment was started with the preparation of a surgical guide, prior to the manipulation of tissues to create the peri-implant papillae. In the surgery, two semilunar incisions are made, joined by a perpendicular incision, which when sutured on them allowed to create the papillae. An implant with good primary stability is installed and the impression is taken for rehabilitation. At 24 h a screwed crown is installed, showing the presence of the papillae immediately. Controls are performed up to 20 months of function, demonstrating good stability of the mesial and distal papillae without presenting any type of complication.

Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA

Abstract Objective The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). Material and Methods Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. Results In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. Conclusion mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Inhibition of autophagy suppresses osteogenic differentiation of stem cells from apical papilla / 南方医科大学学报

OBJECTIVE@#To investigate the effects of autophagy on osteogenic differentiation of stem cells from the apical papilla (SCAPs) in the presence of tumor necrosis factor- (TNF-) stimulation .@*METHODS@#SCAPs treated with TNF- (0, 5, and 10 ng/mL) with or without 5 mmol/L 3-MA were examined for the expression of autophagy marker LC3-Ⅱ using Western blotting. The cells were transfected with GFP-LC3 plasmid and fluorescence microscopy was used for quantitative analysis of intracellular GFP-LC3; AO staining was used to detect the acidic vesicles in the cells. The cell viability was assessed with CCK-8 assays and the cell apoptosis rate was analyzed using flow cytometry. The cells treated with TNF- or with TNF- and 3-MA were cultured in osteogenic differentiation medium for 3 to 14 days, and real- time PCR was used to detect the mRNA expressions of osteogenesis-related genes (ALP, BSP, and OCN) for evaluating the cell differentiation.@*RESULTS@#TNF- induced activation of autophagy in cultured SCAPs. Pharmacological inhibition of TNF--induced autophagy by 3-MA significantly decreased the cell viability and increased the apoptosis rate of SCAPs ( < 0.05). Compared with the cells treated with TNF- alone, the cells treated with both TNF- and 3-MA exhibited decreased expressions of the ALP and BSP mRNA on days 3, 7 and 14 during osteogenic induction ( < 0.05) and decreased expression of OCN mRNA on days 3 and 7 during the induction ( < 0.05).@*CONCLUSIONS@#Autophagy may play an important role during the osteogenic differentiation of SCAPs in the presence of TNF- stimulation.

Root canal dressings for revascularization influence in vitro mineralization of apical papilla cells

Abstract Endodontic revascularization is based on cell recruitment into the necrotic root canal of immature teeth after chemical disinfection. The clinical outcome depends on the ability of surviving cells from the apical tissue to differentiate and promote hard tissue deposition inside the dentinal walls. Objective To investigate the effect of calcium hydroxide (CH) and modified triple antibiotic paste (mTAP - ciprofloxacin, metronidazole and cefaclor) on the viability and mineralization potential of apical papilla cells (APC) in vitro . Material and Methods APC cultures were kept in contact with CH or mTAP (250-1000 µg/mL) for 5 days, after which cell viability was assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Next, APCs were subjected to CH or mTAP at 250 µg/mL for 5 days before inducing the differentiation assay. After 14 and 21 days, calcium deposition was assessed by the Alizarin Red S staining method, followed by elution and quantification using spectrophotometry. Data were analyzed using ANOVA followed by Tukey post hoc test. Results CH induced cell proliferation, whereas mTAP showed significant cytotoxicity at all concentrations tested. APC treated with CH demonstrated improved mineralization capacity at 14 days, while, for mTAP, significant reduction on the mineralization rate was observed for both experimental periods (14 and 21 days). Conclusion Our findings showed that CH induces cell proliferation and improves early mineralization, whereas mTAP was found cytotoxic and reduced the mineralization potential in vitro of APCs.

Extracellular calcium increases fibroblast growth factor 2 gene expression via extracellular signal-regulated kinase 1/2 and protein kinase A signaling in mouse dental papilla cells

Abstract We previously reported that elevated extracellular calcium (Ca2+) levels increase bone morphogenetic protein 2 expression in human dental pulp (hDP) cells. However, it is unknown whether extracellular Ca2+ affects the expression of other growth factors such as fibroblast growth factor 2 (FGF2). Objective: The present study aimed to examine the effect of extracellular Ca2+ on FGF2 gene expression in hDP and immortalized mouse dental papilla (mDP) cells. Materials and Methods: Cells were stimulated with 10 mM CaCl2 in the presence or absence of cell signaling inhibitors. FGF2 gene expression was assessed using real-time polymerase chain reaction. The phosphorylation status of signaling molecules was examined by Western blotting. Results: Extracellular Ca2+ increased FGF2 gene expression in mDP and hDP cells. Gene expression of the calcium-sensing receptor and G protein-coupled receptor family C group 6 member A, both of which are extracellular Ca2+ sensors, was not detected. Ca2+-mediated Fgf2 expression was reduced by pretreatment with the protein kinase A (PKA) inhibitor H-89 or extracellular signal-regulated kinase (ERK) 1/2 inhibitor PD98059 but not by pretreatment with the protein kinase C inhibitor GF-109203X or p38 inhibitor SB203580. Extracellular Ca2+ increased PKA activity and ERK1/2 phosphorylation. Ca2+-induced PKA activity decreased by pretreatment with PD98059. Conclusions: These findings indicate that elevated extracellular Ca2+ levels led to increased Fgf2 expression through ERK1/2 and PKA in mDP cells and that this mechanism may be useful for designing regenerative therapies for dentin.

Angiotensina II na modulação de células de papila apical humana in vitro / Angiotensin II in the modulation of apical human papilla cells in vitro

A Angiotensina II (Ang II), principal peptídeo efetor do Sistema Renina Angiotensina (SRA), tem sido relatada como importante mediador de processos inflamatórios. Considera-se de extrema relevância detectar esse sistema e compreender a modulação da Ang II nas células da papila apical (CPA) considerando que esta população celular é a chave no sucesso na revascularização pulpar. Vale também considerar que a infecção do sistema de canais radiculares pode alterar as funções destas células. Sendo assim, o presente trabalho in vitro teve como objetivo elucidar o papel do lipopolissacarídeo (LPS) nas células da papila apical, detectar os componentes do Sistema Renina Angiotensina (SRA) e compreender a modulação da Ang II nas CPA. Para vencer o desafio científico, culturas de células da papila foram estabelecidas e caracterizadas fenotipicamente por citometria de fluxo e funcionalmente por Vermelho de Alizarina. Quando estimuladas por LPS, foi feita análise da citotoxicidade por MTT e produção de citocinas e Osteoprotegerina (OPG) por ELISA. Em seguida, a detecção da expressão gênica dos componentes do Sistema Renina Angiotensina foi realizada por Transcrição Reversa seguida de Reação em Cadeia da Polimerase quantitativa (RT-qPCR) e do peptídeo Ang II por ELISA. As células foram estimuladas com LPS, a própria Ang II e um antagonista do receptor de Angiotensina II tipo 1 (AT1) e avaliadas quanto à citotoxicidade por ensaio de MTT, capacidade de formação de nódulos mineralizados por Vermelho de Alizarina e produção de citocinas inflamatórias e OPG por ELISA. Os resultados demonstraram que culturas de CPA expressaram níveis típicos de marcadores de células-tronco mesenquimais CD146, CD24 e STR0-1 e formaram nódulos mineralizados com o meio de diferenciação. O estímulo com LPS a 0,1; 1 e 10?g/mL no decorrer de 1, 3, 5, 7 e 14 dias não afetou a viabilidade das CPA. As células estimuladas com LPS apresentaram níveis maiores da quimiocina MCP-1/ CCL-2 a 1?g/mL em uma e 24 horas e a expressão de OPG diminui na presença de LPS a 0,1 e 10?g/mL. Na concentração de 1?g/mL, foi detectada a expressão gênica de Angiotensinogênio, Renina, Enzima Conversora de Angiotensina (ECA) e AT1. Houve maior expressão de ECA no período experimental de 1 hora com LPS e em 24 horas, apenas AT1 e ECA continuaram presentes. Sendo a produção da proteína soberana, Ang II foi encontrada nas células pelo ELISA. Os ensaios funcionais demonstraram que a Ang II aumentou a proliferação celular em 24, 48 e 72 horas. Em 24 horas, a presença do peptídeo efetor do SRA aumentou a produção da quimiociona MCP-1/ CCL-2 e do ligante do ativador do receptor do fator nuclear- ?B (RANKL). Concluímos que o LPS foi capaz de alterar funcionalmente as CPA, que o Sistema Renina Angiotensina está presente e a Ang II tem capacidade de modulação pró-inflamatória, proliferativa e de reabsorção óssea nesta população celular.

Effect of Resveratrol on Cell Differentiation and Mineralization in Cultured Odontoblasts

Resveratrol (3,4′,5,-trihydroxystilbene), a phytoalexin present in grapes, exerts a variety of actions to reduce superoxides, prevents diabetes mellitus, and inhibits inflammation. Resveratrol acts as a chemo-preventive agent and induces apoptotic cell death in various cancer cells. However, the role of resveratrol in odontoblastic cell differentiation is unclear. In this study, the effect of resveratrol on regulating odontoblast differentiation was examined in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. Resveratrol significantly accelerated mineralization as compared with the control culture in differentiation of MDPC-23 cells. Resveratrol significantly increased expression of ALP mRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly accelerated expression of ColImRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly increased expressions of DSPP and DMP-1 mRNAs as compared with the control in differentiation of MDPC-23 cells. Treatment of resveratrol did not significantly affect cell proliferation in MDPC-23 cells. Results suggest resveratrol facilitates odontoblast differentiation and mineralization in differentiation of MDPC-23 cells, and may have potential properties for development and clinical application of dentin regeneration materials.

Expressão gênica de BMPR1, BMPR2 e TGFBR1 liberados de discos de dentina humana tratados com diferentes soluções desmineralizantes e adesão das células da papila apical cultivadas / Gene expression of BMPR1, BMPR2 and TGFBR1 released from human dentin disks treated with different demineralizing solutions and adhesion of cultured apical papilla cells

O sucesso dos procedimentos endodônticos regenerativos depende da adequada descontaminação do espaço endodôntico, da presença de células mesenquimais indiferenciadas, da liberação de fatores de crescimento que atuam como moléculas sinalizadoras para atração, proliferação, diferenciação celular e da presença de um "scaffold" que propicia suporte para a organização e vascularização do tecido neoformado. Os protocolos de regeneração pulpar têm utilizado o EDTA (ácido etileno-diamino-tetracético) para remover o smear layer e liberar fatores de crescimento presentes na matriz dentinária. Poucos estudos analisaram se a utilização de soluções quelantes específicas para o cálcio ou menos agressivas, apresentam melhores resultados na liberação de fatores de crescimento e na adesão das células da papila apical à superfície dentinária. O objetivo deste estudo foi avaliar a expressão gênica de BMPR1, BMPR2 e TGFBR1 liberados de discos de dentina humana previamente tratados com EDTA, EGTA e Ácido Cítrico e a adesão das células da papila apical à esses discos. Foram utilizadas células criopreservadas do biorrepositório de células do Laboratório de Biologia Básica do Departamento de Dentística da Faculdade de Odontologia da Universidade de São Paulo. Os discos de dentina foram obtidos da porção radicular interna de pré-molares e terceiros molares hígidos, humanos, que foram obtidos do Biobanco da FOUSP - Divisão de Dentes Humanos. Após a imersão por 1 minuro nas soluções desmineralizantes, os discos de detina foram transferidos para placas com 90 poços e as células foram plaqueadas. A adesão celular foi avaliada após 48 horas empregando-se a Microscopia Eletrônica de Varedura. Determinou-se a expressão gênica após os períodos de 07 e 21 dias de cultivo celular. A análise estatística foi realizada por meio da análise de variância a 1 critério (ANOVA) seguida de pós teste de Tukey, com nível de significância de 5%. Aos 21 dias o tratamento da dentina com EGTA resultou em significativo aumento da BMPR1, comparado com PBS. Nenhuma condição experimental alterou a expressão de BMPR2. A expressão da TGBR1 aos 21 dias foi significativamente aumentada pelo tratamento prévio dos discos com EGTA. Embora não tenha ocorrido distribuição uniforme de células ao longo dos discos, foi observada adesão celular em todos os grupos experimentais. Conclui-se que o tipo de solução desmineralizante interfere na quantidade liberada dos fatores de crescimento BMPR1 e TGFBR1, presentes na dentina humana, não interferindo na adesão das células da papila apical na superfície dentinária.

Effect of Metformin on Cell Growth and Differentiation in Cultured Odontoblasts

Metformin (1,1-dimethylbiguanide hydrochloride), derived from French lilac (Galega officinalis), is a first-line anti-diabetic drug prescribed for patients with type 2 diabetes. However, the role of metformin in odontoblastic cell differentiation is still unclear. This study therefore undertook to examine the effect of metformin on regulating odontoblast differentiation in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. As compared to controls, metformin significantly accelerated the mineralization, significantly increased and accelerated the expressions of ALP and Col I mRNAs, and significantly increased the accelerated expressions of DSPP and DMP-1 mRNAs, during differentiation of MDPC-23 cells. There was no alteration in cell proliferation of MDPC-23 cells, on exposure to metformin. These results suggest that the effect of metformin on MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells, facilitates the odontoblast differentiation and mineralization, without altering the cell proliferation.

Tratamiento de recesión única de clase III con injerto libre subepitelial mediante técnica de colgajo de doble papila reposicionado: informe de un caso clínico / Treatment of a single Class III recession combining free subepithelial connective tissue graft and a double papilla repositioned flap: report of a clinical case

Objetivo: mostrar y evaluar los resultados clínicos de un procedimiento de cirugía plástica periodontal, efectuada para cubrir una recesión radicular de clase III de Miller, con una técnica bilaminar. Además, analizar el aumento de la banda de encía y el cambio de biotipo gingival de la pieza dentaria. Caso clínico: paciente femenino de 30 años de edad, con una recesión gingival de Clase III de Miller por vestibular de la pieza 43 que no sobrepasa la línea mucogingival, con pérdida interproximal de tejidos duros y blandos. El tratamiento consiste en un colgajo de doble papila a espesor parcial, con injerto libre subepitelial tomado del paladar, con seguimiento a 1 año. Conclusiones: la técnica bilaminar es una solución viable en casos de recubrimiento radicular poco predecibles, como la recesión de clase III de Miller. El biotipo gingival se vio engrosado y la encía queratinizada no sufrió variaciones.

Induced Pluripotent Stem (iPS) Cells in Dentistry: A Review

iPS cells are derived from somatic cells via transduction and expression of selective transcription factors. Both viral-integrating (like retroviral) and non-integrating (like, mRNA or protein-based) techniques are available for the production of iPS cells. In the field of dentistry, iPS cells have been derived from stem cells of apical papilla, dental pulp stem cells, and stem cells from exfoliated deciduous teeth, gingival and periodontal ligament fibroblasts, and buccal mucosa fibroblasts. iPS cells have the potential to differentiate into all derivatives of the 3 primary germ layers i.e. ectoderm, endoderm, and mesoderm. They are autogeneically accessible, and can produce patient-specific or disease-specific cell lines without the issue of ethical controversy. They have been successfully tested to produce mesenchymal stem cells-like cells, neural crest-like cells, ameloblasts-like cells, odontoblasts-like cells, and osteoprogenitor cells. These cells can aid in regeneration of periodontal ligament, alveolar bone, cementum, dentin-pulp complex, as well as possible Biotooth formation. However certain key issues like, epigenetic memory of iPS cells, viral-transduction, tumorgenesis and teratoma formation need to be overcome, before they can be successfully used in clinical practice. The article discusses the sources, pros and cons, and current applications of iPS cells in dentistry with an emphasis on encountered challenges and their solutions.

Requirement of Smad4-mediated signaling in odontoblast differentiation and dentin matrix formation / 대한해부학회지

Dentin is the major part of tooth and formed by odontoblasts. Under the influence of the inner enamel epithelium, odontoblasts differentiate from ectomesenchymal cells of the dental papilla and secrete pre-dentin which then undergo mineralization into dentin. Transforming growth factor-beta (TGF-β)/bone morphogenetic protein (BMP) signaling is essential for dentinogenesis; however, the precise molecular mechanisms remain unclear. To understand the role of TGF-β/BMP signaling in odontoblast differentiation and dentin formation, we generated mice with conditional ablation of Smad4, a key intracellular mediator of TGF-β/BMP signaling, using Osr2 or OC-Cre mice. Here we found the molars of Osr2(Cre)Smad4 mutant mice exhibited impaired odontoblast differentiation, and normal dentin was replaced by ectopic bone-like structure. In Osr2(Cre)Smad4 mutant mice, cell polarity of odontoblast was lost, and the thickness of crown dentin was decreased in later stage compared to wild type. Moreover, the root dentin was also impaired and showed ectopic bone-like structure similar to Osr2(Cre)Smad4 mutant mice. Taken together, our results suggest that Smad4-dependent TGF-β/BMP signaling plays a critical role in odontoblast differentiation and dentin formation during tooth development.

Extrusão ortodôntica para restabelecer a arquitetura gengival / Orthodontic extrusion for restoring the gingival architecture: a clinical case report

Este artigo procurou enfatizar a importância de se realizar uma adequada avaliação tecidual antes de iniciar reabilitações estéticas, além de sugerir possíveis condutas para restabelecer a arquitetura gengival. No caso clínico apresentado, os incisivos centrais superiores foram tracionados lentamente por aparatologia ortodôntica, visando nivelar o arco gengival e devolver a papila interdental. Após a obtenção de um quadro gengival mais harmônico, a forma dental foi restaurada com facetas cerâmicas para alcançar um bom resultado estético.

This article emphasizes the importance of adequate tissue assessment before starting aesthetic restorations, as well as to suggest possible guidelines to restore the gingival architecture. In the clinical case presented, the upper central incisors were orthodontically extruded, aiming to level the gingival arch and to retrieve the dental papillae. After obtaining a more harmonic gingival architecture, the dental form was restored with ceramic veneers to achieve good aesthetic result.

Papilla Reconstruction: A Case Report.

Smile plays an important role in improving esthetics and radiates health and self-confidence. The perfect smile requires an optimal relationship between the teeth, surrounding oral tissues, and periodontal complex. When a disharmony exists between any of these components, the result is a smile that is likely to be perceived as unaesthetic. Similar is the case with interdental bone loss resulting in increased embrasure spaces, with unaesthetic appearance especially in the anterior maxillary regions. Periodontal papilla reconstruction has proved to be a successful technique and a boon to the patient in such a case to return an esthetic and healthy smile to the patient. We report here a similar case of papilla reconstruction with pre-treatment and post-treatment differences depicted in the esthetics.