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Article in English | WPRIM | ID: wpr-880855


Dental pulp can initiate its damage repair after an injury of the pulp-dentin complex by rearrangement of odontoblasts and formation of newly differentiated odontoblast-like cells. Connexin 43 (Cx43) is one of the gap junction proteins that participates in multiple tissue repair processes. However, the role of Cx43 in the repair of the dental pulp remains unclear. This study aimed to determine the function of Cx43 in the odontoblast arrangement patterns and odontoblastic differentiation. Human teeth for in vitro experiments were acquired, and a pulp injury model in Sprague-Dawley rats was used for in vivo analysis. The odontoblast arrangement pattern and the expression of Cx43 and dentin sialophosphoprotein (DSPP) were assessed. To investigate the function of Cx43 in odontoblastic differentiation, we overexpressed or inhibited Cx43. The results indicated that polarized odontoblasts were arranged along the pulp-dentin interface and had high levels of Cx43 expression in the healthy teeth; however, the odontoblast arrangement pattern was slightly changed concomitant to an increase in the Cx43 expression in the carious teeth. Regularly arranged odontoblast-like cells had high levels of the Cx43 expression during the formation of mature dentin, but the odontoblast-like cells were not regularly arranged beneath immature osteodentin in the pulp injury models. Subsequent in vitro experiments demonstrated that Cx43 is upregulated during odontoblastic differentiation of the dental pulp cells, and inhibition or overexpression of Cx43 influence the odontoblastic differentiation. Thus, Cx43 may be involved in the maintenance of odontoblast arrangement patterns, and influence the pulp repair outcomes by the regulation of odontoblastic differentiation.

Animals , Cell Differentiation , Connexin 43 , Dental Pulp , Extracellular Matrix Proteins , Odontoblasts , Phosphoproteins , Rats , Rats, Sprague-Dawley
Odontoestomatol ; 23(38): e207, 2021. graf
Article in Spanish | LILACS, BNUY, BNUY-Odon | ID: biblio-1340273


Resumen Objetivos: Establecer e implementar un protocolo simplificado de extracción, aislamiento primario y cultivo de células madre derivadas de la pulpa dental humana (DPSCh). Analizar cuantitativamente y cualitativamente las células aisladas. Metodología: 10 terceros molares sanos donados por pacientes que concurrieron a la Facultad de Odontología, UdelaR y otorgaron su consentimiento escrito fueron procesados antes de las 48 hs. Se realizó la fractura de la pieza para la obtención del tejido pulpar y se procesó por el método explante. Se analizó viabilidad celular y expresión de marcadores por citometría de flujo en pasajes 4 y 12 y se corroboró mediante inmunocitoquímica. Resultados: Las células obtenidas presentaron una vitalidad mayor al 90% en todos los pasajes, observándose una morfología característica y expresión de marcadores de células madre mesenquimales CD90, C105, CD73, CD29 y 166 mediante citometría de flujo en ambos pasajes. Conclusiones: Se logró establecer un protocolo de aislamiento y expansión celular, con alta tasa de éxito de una población de DPSCh.

Resumo Objetivos: Estabelecer e implementar um protocolo simplificado para a extração, isolamento primário e cultura de células-tronco da polpa dentária humana (DPSCh). Analise as células isoladas quantitativa e qualitativamente. Metodologia: 10 terceiros molares saudáveis ​​doados por pacientes que frequentaram a Faculdade de Odontologia UdelaR e deram consentimento por escrito foram processados ​​antes de 48 horas. A fratura da peça foi realizada para obtenção do tecido pulpar e processada pelo método do explante. A viabilidade celular e a expressão do marcador foram analisadas por citometría de fluxo nas passagens 4 e 12 e confirmadas por inmunocitoquímica. Resultados: As células obtidas apresentaram viabilidade superior a 90% em todas as passagens, observando uma morfologia característica e expressão dos marcadores de células-tronco mesenquimais CD90, C105, CD73, CD29 e 166 por citometría de fluxo em ambas as passagens. Conclusões: Foi possível estabelecer um protocolo de isolamento celular, com alta taxa de sucesso e segurança para isolar o DPSCh.

Abstract Objectives: To establish and implement a simplified protocol for the extraction, primary isolation, and culture of human dental pulp stem cells (hDPSCs). To analyze the isolated cells quantitatively and qualitatively. Methodology: Ten healthy third molars were donated by patients who attended the School of Dentistry, UdelaR, and gave their written consent. The teeth were processed within 48 hours. The teeth were sectioned to obtain the pulp tissue and processed with the explant method. Cell viability and marker expression were analyzed by flow cytometry at passages 4 and 12 and verified by immunocytochemistry. Results: The cells obtained had a vitality greater than 90% in all passages. We found the characteristic morphology and the expression of CD90, C105, CD73, CD29 and 166 mesenchymal stem cell markers by flow cytometry in both passages. Conclusion: It was possible to establish a cell isolation protocol that is highly successful and safe to isolate hDPSC.

Cell Separation , Laboratory Equipment , Tissue and Organ Harvesting/methods , Adult Stem Cells , Flow Cytometry , Dental Pulp , Mesenchymal Stem Cells
J. appl. oral sci ; 29: e20201058, 2021. graf
Article in English | LILACS | ID: biblio-1286914


Abstract Apical periodontitis is an inflammatory disorder of periradicular tissues developed from endodontic infections. Understanding its pathophysiology and the underlying molecular mechanisms is key to the advancement of endodontics. MicroRNAs (miRNAs), a group of evolutionarily conserved small non-coding RNAs, may be phenotypically and functionally associated with the pathogenesis of apical periodontitis. Several studies have focused on the role of miRNAs in the pulp and periradicular biology, and they have demonstrated their essential functions, such as initiating odontogenic differentiation and promoting pro- or anti-inflammatory responses in pulpitis. Up to date, over 2,000 miRNAs have been discovered in humans; however, only few have been reported to associate with apical periodontitis. Therefore, identifying miRNAs involved in diseased apical tissues and conducting functional studies are important in expanding our current knowledge of pulp and periradicular biology and exploring novel therapeutic avenues. In this review, we revisit current models of apical periodontitis and miRNA biogenesis, analyze existing evidence of the involvement of miRNAs in diseased apical tissues, and discuss their diverse functions and potential values. Based on their sheer abundance, prolonged stability in biofluid, and relative ease of sampling, miRNAs may be a useful tool to be developed as diagnostic biomarkers for apical periodontitis. Furthermore, it can be used as therapeutic targets in conjunction with conventional endodontic therapies.

Humans , Periapical Periodontitis/genetics , Pulpitis , MicroRNAs/genetics , Endodontics , Dental Pulp
J. appl. oral sci ; 29: e20200998, 2021. tab, graf
Article in English | LILACS | ID: biblio-1286921


Abstract Objectives The aim of this study is to evaluate the effect of using gel and solution forms of NaOCI during the chemomechanical preparation of the root canals on postoperative pain at different time intervals. Methodology 114 patients with mandibular molar teeth and symptomatic irreversible pulpitis were included in the study. All patients were divided into two groups based on the irrigant used during root canal preparation (n=57): Group 1, 5.25% NaOCI, Group 2, 5.25% NaOCI gel. All groups were filled with gutta-percha and AH Plus root canal sealer using single-cone technique. VAS scale (1-10) was used for postoperative pain assessment. After endodontic treatment, all patients were asked to record their postoperative pain levels at the 6th, 24th, 48th, 72nd hours, and 1 week later. The data were analyzed using Chi-Squared, Independent Samples T, Cochran Q and Friedman tests. Results Statistically significant difference was not found between the distributions of pain levels at different times according to the groups (p>0.050). A statistically significant difference was observed between the distributions of pain levels measured at different times in the solution group (p<0.001). A statistically significant difference was found between the distributions of pain levels measured at different times in the gel group (p<0.001). In both groups, highest postoperative pain levels occurred in the first 6 hours. Pain levels of the gel group as 38,5% mild, 17.3% moderate, 5.8% severe and pain levels of the solution group were obtained as 46.2% mild, 26.9% moderate, 9.6% severe at the 6th hour. Conclusions The use of the gel form of NaOCI during the chemomechanical preparation of the root canals showed similar postoperative pain when compared to the solution form.

Humans , Root Canal Filling Materials , Sodium Hypochlorite , Pain, Postoperative/prevention & control , Root Canal Obturation , Root Canal Preparation , Dental Pulp , Dental Pulp Cavity , Molar
J. appl. oral sci ; 29: e20201074, 2021. tab, graf
Article in English | LILACS | ID: biblio-1340110


Abstract Hyperglycemia, a major characteristic of diabetes, is considered to play a vital role in diabetic complications. High glucose levels have been found to inhibit the mineralization of dental pulp cells. However, gene expression associated with this phenomenon has not yet been reported. This is important for future dental therapeutic application. Objective Our study aimed to investigate the effect of high glucose levels on mineralization of human dental pulp-derived cells (hDPCs) and identify the genes involved. Methodology hDPCs were cultured in mineralizing medium containing 25 or 5.5 mM D-glucose. On days 1 and 14, RNA was extracted and expression microarray performed. Then, differentially expressed genes (DEGs) were selected for further validation using the reverse transcription quantitative polymerase chain reaction (RT-qPCR) method. Cells were fixed and stained with alizarin red on day 21 to detect the formation of mineralized nodules, which was further quantified by acetic acid extraction. Results Comparisons between high-glucose and low-glucose conditions showed that on day 1, there were 72 significantly up-regulated and 75 down-regulated genes in the high-glucose condition. Moreover, 115 significantly up- and 292 down-regulated genes were identified in the high-glucose condition on day 14. DEGs were enriched in different GO terms and pathways, such as biological and cellular processes, metabolic pathways, cytokine-cytokine receptor interaction and AGE-RAGE signaling pathways. RT-qPCR results confirmed the significant expression of pyruvate dehydrogenase kinase 3 (PDK3), cyclin-dependent kinase 8 (CDK8), activating transcription factor 3 (ATF3), fibulin-7 (Fbln-7), hyaluronan synthase 1 (HAS1), interleukin 4 receptor (IL-4R) and apolipoprotein C1 (ApoC1). Conclusions The high-glucose condition significantly inhibited the mineralization of hDPCs. DEGs were identified, and interestingly, HAS1 and Fbln-7 genes may be involved in the glucose inhibitory effect on hDPC mineralization.

Humans , Dental Pulp , Transcriptome , Cell Differentiation , Cells, Cultured , Microarray Analysis , Cell Proliferation , Glucose
J. appl. oral sci ; 29: e20210296, 2021. graf
Article in English | LILACS | ID: biblio-1340101


Abstract Objectives Human dental pulp stem cells (DPSCs) have been used to regenerate damaged nervous tissues. However, the methods of committing DPSCs into neural stem/progenitor cells (NSPCs) or neurospheres are highly diverse, resulting in many neuronal differentiation outcomes. This study aims to validate an optimal protocol for inducing DPSCs into neurospheres and neurons. Methodology After isolation and characterization of mesenchymal stem cell identity, DPSCs were cultured in a NSPC induction medium and culture vessels. The durations of the culture, dissociation methods, and passage numbers of DPSCs were varied. Results Neurosphere formation requires a special surface that inhibits cell attachment. Five-days was the most appropriate duration for generating proliferative neurospheres and they strongly expressed Nestin, an NSPC marker. Neurosphere reformation after being dissociated by the Accutase enzyme was significantly higher than other methods. Passage number of DPSCs did not affect neurosphere formation, but did influence neuronal differentiation. We found that the cells expressing a neuronal marker, β-tubulin III, and exhibiting neuronal morphology were significantly higher in the early passage of the DPSCs. Conclusion These results suggest a guideline to obtain a high efficiency of neurospheres and neuronal differentiation from DPSCs for further study and neurodegeneration therapeutics.

Humans , Stem Cells , Dental Pulp , Cell Differentiation
Rev. Fac. Odontol. (B.Aires) ; 36(83): 67-74, 2021. ilus
Article in Spanish | LILACS | ID: biblio-1343747


El presente trabajo de investigación tiene como objetivo principal el aislar, expandir y caracterizar inmunofenotípicamente células madre mesenquimales de la pulpa dental humana, según los criterios mínimos propuestos por The International Society for Cellular Therapy (ISCT), como así también establecer la puesta a punto de las técnicas y protocolos de procedimientos para tal fin. Los cultivos fueron permanentemente monitoreados mediante microscopio invertido con contraste de fase y la inmunotipificación fue realizada por citometría de flujo (AU)

Humans , Male , Female , Tissue Engineering , Dental Pulp , Adult Stem Cells , Mesenchymal Stem Cells , Phenotype , Argentina , Schools, Dental , Cell Culture Techniques , Regenerative Medicine
Braz. oral res. (Online) ; 35: e004, 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132741


Abstract: There is a lack of evidence about the best approach for cavitated caries lesions with the possibility of pulpal involvement in primary teeth. Thus, the present authors aimed to verify the best treatment for deep caries lesions with or without pulp involvement in primary teeth. The search was conducted in MEDLINE/Pubmed and Web of Science databases until May 2020. Studies that compared techniques to manage deep caries lesions with at least 12 months of follow-up were included. The risk of bias was evaluated using the RoB tool. Network meta-analysis and pairwise meta-analyses were conducted considering the treatment clinical success as an outcome, according to the pulp health condition. From 491 potentially eligible studies, 9 were included. For deep caries lesions with pulp vitality, the Hall Technique presented the highest probability of success (78%). In the event of accidental pulp exposure, pulpectomy presented a 76% chance of providing the best clinical results. For pulp necrosis, no difference was observed between a pulpectomy and non-instrumented endodontic treatment (RR = 0.69; 95%CI: 0.21-2.33) Thus, it was concluded that the Hall Technique may be a better option for deep caries lesions with pulp vitality. In cases of accidental pulp exposure of vital teeth during caries removal, a pulpectomy may be considered the best option. However, there are insufficient studies to build up evidence about the best treatment option when irreversible pulpitis or pulp necrosis is present.

Humans , Tooth, Deciduous , Dental Caries/therapy , Pulpectomy , Dental Pulp , Network Meta-Analysis
Article in Chinese | WPRIM | ID: wpr-879535


OBJECTIVE@#To compare the mRNA level of cell proliferation-related genes Twist1, SIRT1, FGF2 and TGF-β3 in placenta mesenchymal stem cells (PA-MSCs), umbilical cord mensenchymals (UC-MSCs) and dental pulp mesenchymal stem cells (DP-MSCs).@*METHODS@#The morphology of various passages of PA-MSCs, UC-MSCs and DP-MSCs were observed by microscopy. Proliferation and promoting ability of the three cell lines were detected with the MTT method. Real-time PCR (RT-PCR) was used to determine the mRNA levels of Twist1, SIRT1, FGF2, TGF-β3.@*RESULTS@#The morphology of UC-MSCs and DP-MSCs was different from that of PA-MSCs. Proliferation ability and promoting ability of the PA-MSCs was superior to that of UC-MSCs and DP-MSCs. In PA-MSCs, expression level of Twist1 and TGF-β3 was the highest and FGF2 was the lowest. SIRT1 was highly expressed in UC-MSCs. With the cell subcultured, different expression levels of Twist1, SIRT1, FGF2, TGF-β3 was observed in PA-MSCs, UC-MSCs and DP-MSCs.@*CONCLUSION@#Up-regulated expression of the Twist1, SIRT1 and TGF-β3 genes can promote proliferation of PA-MSCs, UC-MSCs and DP-MSCs, whilst TGF-β3 may inhibit these. The regulatory effect of Twist1, SIRT1, FGF2 and TGF-β3 genes on PA-MSCs, UC-MSCs and DP-MSCs are different.

Cell Differentiation , Cell Proliferation/genetics , Cells, Cultured , Dental Pulp/cytology , Female , Fibroblast Growth Factor 2/genetics , Humans , Mesenchymal Stem Cells/cytology , Nuclear Proteins/genetics , Placenta/cytology , Pregnancy , Sirtuin 1/genetics , Transforming Growth Factor beta3/genetics , Twist-Related Protein 1/genetics , Umbilical Cord/cytology
Int. j. morphol ; 38(6): 1742-1750, Dec. 2020. tab, graf
Article in English | LILACS | ID: biblio-1134507


SUMMARY: Mesenchymal stem cells are present in adult tissues such as the human dental pulp. They are pluripotent and can differentiate into various specialized cell types in vitro through appropriate stimuli. Ameloblasts produce human tooth enamel only during embryonic development before tooth eruption, so endogenous regeneration is not possible. Various efforts have been aimed at generating natural or artificial substitutes for dental enamel with properties similar to the specific components of said tissue. The purpose of this study was to induce human dental pulp stem cells to produce enamel proteins using extracellular matrix derived from the rat tail tendon and pigskin. Primary cultures of human dental pulp stem cells were established and characterized by RT-PCR and immunofluorescence, using mesenchymal cell markers such as CD14, CD40, CD44, CD105, and STRO-1. The cells were then incubated with the extracellular matrix for fourteen days and labeled with specific antibodies to detect the expression of dental enamel proteins such as amelogenin, ameloblastin, enamelisin, tuftelin, and parvalbumin, characteristics of the phenotype of ameloblasts. This work demonstrated a positive effect of the extracellular matrix to induce the expression of enamel proteins in the stem cells of the human dental pulp.

RESUMEN: Las células madre mesenquimales están presentes en los tejidos adultos como la pulpa dental humana. Son pluripotentes y pueden diferenciarse en varios tipos de células especializadas in vitro a través de estímulos adecuados. Los ameloblastos producen esmalte dental humano sólo durante el desarrollo embrionario antes de la erupción dental, por lo que no es posible su regeneración endógena. Varios esfuerzos se han orientado a generar sustitutos naturales o artificiales de esmalte dental con propiedades similares a los componentes específicos de este tejido. El propósito de este estudio fue inducir células madre de pulpa dental humana para producir proteínas del esmalte dental a través del estímulo de matriz extracelular derivada del tendón de la cola de rata y piel de cerdo. Se establecieron cultivos primarios de células madre de pulpa dental humana y se caracterizaron por RT-PCR e inmunofluorescencia utilizando marcadores de células mesenquimales como CD14, CD40, CD44, CD105 y STRO-1. Posteriormente, las células se incubaron con matriz extracelular durante un período de catorce días y se marcaron con anticuerpos específicos para detectar la expresión de proteínas de esmalte dental como amelogenina, ameloblastina, enamelisina, tuftelina y parvalbúmina, las cuales son características del fenotipo de ameloblastos. Este trabajo demostró el efecto positivo que tiene el empleo de la matriz extracelular para inducir la expresión de proteínas de esmalte en las células pluripotenciales de la pulpa dental humana.

Humans , Stem Cells , Dental Enamel Proteins , Dental Pulp , Extracellular Matrix , Immunophenotyping , Fluorescent Antibody Technique , Cell Culture Techniques , Tissue Engineering
Dent. press endod ; 10(2): 73-78, maio-ago.2020. Ilus
Article in English | LILACS | ID: biblio-1344662


Introdução: Reabsorção radicular interna é a perda progressiva de dentina intrarradicular como resultado de atividades clásticas. Geralmente, mantém-se assintomática e é diagnosticada durante o exame radiográfico de rotina. O correto diagnóstico é de extrema importância para evitar a terapêutica inadequada, já que, em alguns casos, sua progressão pode resultar na perda do elemento dentário. Objetivo: O objetivo do presente trabalho é relatar um caso clínico de perfuração radicular causada por uma reabsorção interna inflamatória em incisivo central superior, em terço médio da raiz, com integridade do terço apical. Métodos: O tratamento foi realizado usando técnica de obturação com cones de guta-percha e cimento endodôntico à base de resina epóxi no terço apical e agregado trióxido mineral na área da reabsorção, com auxílio do microscópio operatório. Resultado: Após um período de três anos de acompanhamento clínico e radiográfico, pode-se confirmar o processo de reparo, demonstrando que é possível obter sucesso em casos de reabsorção perfurante. Conclusão: Mesmo em casos em que há comunicação da polpa dentária com tecidos periodontais, é possível salvar o elemento dentário com o diagnóstico preciso e tratamento correto.

Introduction: Internal root resorption is the progressive destruction of intraradicular dentin as a result of clastic activities. It is usually asymptomatic and discovered by chance on routine radiographic examinations. Is of vital importance to make a correct diagnosis so that the appropriate treatment can be provided because in some cases can result in destruction of the dental element. Objective: The aim of this study is to report a case of successful non-surgical management of perforating internal resorption in an maxillary central incisor, in the middle third of the root. Materials and Methods: The treatment was successfully performed with the aid of an operative microscope. The apical third was filled with guttapercha and epoxy-resin-based sealer and the perforation site it was repaired with mineral trioxide aggregate. Conclusion: Clinical findings and periapical radiographs show that the symptoms and signs ceased, and the results were satisfactory at 3-years follow-up with radiographic examination (AU).

Root Canal Filling Materials , Root Resorption , Radiography, Dental , Dental Pulp , Root Canal Obturation , Therapeutics , Perforant Pathway
Rev. cient. Esc. Univ. Cienc. Salud ; 7(1): 22-28, ene.-jun. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1223261


Según la Organización Mundial dela Salud (OMS), entre el 60 y 90% de la población infantil presenta lesiones cariosas concavitación. Las patologías pulpares son consecuencia de la evolución de la caries o traumatismo dental, manifestándose con dolor, inflamación o infección, que obliga a los pacientesa acudir de forma urgente a la consulta odontológica con cuadros de pulpitis reversible, irreversible o necrosis pulpar. Dependiendo dela gravedad de la patología, esta puede intervenirse mediante terapias curativas y cuando ha alcanzado un nivel muy avanzado, laúnica opción es la exodoncia, dejando secuelas a corto, mediano y largo plazo en el niño. Objetivo: Analizar las diferentes patologías pulpares en molares de ciduos de pacientes infantiles entre 5 y 9 años que acuden a la clínica de Odontopediatría de la Facultad de Odontología de la Universidad Nacional Autónoma de Honduras (UNAH) durante 2016-2018.Pacientes y métodos: Estudio descriptivo, retrospectivo y cuantitativo.Se recolectaron historias clínicas de niños entre 5 y 9 años que acudieron entre 2014 -2016 con una muestra de 310 expedientes de un universo de1605. Resultados: Predominaron las patologías pulpares en el género masculino (54.2%). La caries dental fue la etiología más registrada (77.34%),predominó la pulpitis reversible (9.3%), el órgano dentario más afectado, en el sistema de nomenclatura FDI, (Federation Dentaire Internationale), fue el primer molar deciduo inferior izquierdo (7,4). El tratamiento más realizado fue pulpotomía (15.2%). Conclusión: en la población infantil la caries dental no tratada evolucionó en su mayoría apulpitis reversible...(AU)

Humans , Male , Female , Child, Preschool , Child , Pulpitis , Dental Caries/diagnosis , Tooth Injuries , Dental Pulp
Braz. dent. j ; 31(3): 298-303, May-June 2020. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132307


Abstract Diabetes is a group of metabolic disorders that can lead to damage and dysfunction of many organs including the dental pulp. Increased inflammatory response, reduction of dentin formation and impaired healing were reported in diabetic dental pulp. Hyperglycemia, which is a main characteristic of diabetes, was suggested to play a role in many diabetic complications. Therefore our aim was to investigate the effects of high glucose levels on proliferation, reactive oxygen species (ROS) production and odontogenic differentiation of human dental pulp cells (HDPCs). HDPCs were cultured under low glucose (5.5mM Glucose), high glucose (25 mM Glucose) and mannitol (iso-osmolar control) conditions. Cell proliferation was analyzed by MTT assay for 11 days. Glutathione and DCFH-DA assay were used to assess ROS and antioxidant levels after 24 h of glucose exposure. Odontogenic differentiation was evaluated and quantified by alizarin red staining on day 21. Expression of mineralization-associated genes, which were alkaline phosphatase, dentin sialophosphoprotein and osteonectin, was determined by RT-qPCR on day 14. The results showed that high glucose concentration decreased proliferation of HDPCs. Odontogenic differentiation, both by gene expression and mineral matrix deposit, was inhibited by high glucose condition. In addition, high DCF levels and low reduced glutathione levels were observed in high glucose condition. However, no differences were observed between mannitol and low glucose conditions. In conclusion, the results clearly showed the negative effect of high glucose condition on HDPCs proliferation and differentiation. Moreover, it also induced ROS production of HDPCs.

Resumo O diabetes abrange um grupo de distúrbios metabólicos que podem levar a danos e disfunções de muitos órgãos, incluindo a polpa dentária. Aumento da resposta inflamatória, redução da formação de dentina e comprometimento da cicatrização foram relatados na polpa dentária diabética. A hiperglicemia, que é uma característica determinante do diabetes, desempenha um papel importante em muitas complicações diabéticas. Portanto, nosso objetivo foi investigar os efeitos dos altos níveis de glicose na proliferação, produção de espécies reativas de oxigênio (ROS, em inglês) e diferenciação odontogênica das células da polpa dental humana (HDPCs, em inglês). As HDPCs foram cultivadas em condições de baixa glicose (glicose 5,5 mM), alta glicose (glicose 25 mM) e manitol (controle iso-osmolar). A proliferação celular foi analisada pelo ensaio MTT por 11 dias. Glutationa e DCFH-DA foram utilizados para avaliar os níveis de ROS e antioxidantes após 24 h de exposição à glicose. A diferenciação odontogênica foi avaliada e quantificada pela coloração com vermelho de alizarina no dia 21. A expressão de genes associados à mineralização, que eram fosfatase alcalina, sialofosfoproteína de dentina e osteonectina, foi determinada por RT-qPCR no dia 14. Os resultados mostraram que a alta concentração de glicose diminuiu a proliferação de HDPCs. A diferenciação odontogênica, tanto pela expressão gênica quanto pelo depósito da matriz mineral, foi inibida pela condição de alta glicose. Além disso, altos níveis de DCF e níveis reduzidos de glutationa foram observados na condição de alta glicose. No entanto, não foram observadas diferenças entre o manitol e as condições de baixa glicose. Em conclusão, os resultados mostraram claramente o efeito negativo da condição de alta glicose na proliferação e diferenciação de HDPCs. Além disso, essa condição também induziu a produção de ROS em HDPCs.

Humans , Dental Pulp , Alkaline Phosphatase , Phosphoproteins , Cell Differentiation , Cells, Cultured , Extracellular Matrix Proteins , Reactive Oxygen Species , Cell Proliferation , Glucose , Odontoblasts
Braz. dent. j ; 31(3): 244-251, May-June 2020. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132303


Abstract This in vitro study evaluated cell viability and metabolism, nitric oxide release and production of two chemokines and one cytokine by cultured human dental pulp fibroblasts (HDPF) in contact with two glass ionomer cements (Ketac Molar-KM and Vitrebond-VB), Single Bond (SB) and calcium hydroxide (Dycal-DY). Cultures of HDPF were established by means of an explant technique. The specimens were prepared under sterile conditions and in disks measuring 5 mm x 2 mm obtained from a prefabricated mold and placed on a permeable membrane to avoid direct contact with the cells. Cytotoxicity was assessed by Trypan Blue exclusion method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide release in cell supernatant was detected by the Griess Method whereas stromal derived factor-1 alpha (SDF-1α or CXCL12), chemokine (C-X-C motif) ligand 8 [Interleukin 8 (IL-8 or CXCL8)] and interleukin-6 (IL-6) were detected by ELISA. RT-qPCR was employed for gene expression analysis. Statistical analyses were performed by One-way ANOVA followed by Tukey's post hoc test for materials independent of the time, and Two-way ANOVA followed by Bonferroni correction test for the comparisons between materials and experimental time (p<0.05). Cytotoxic tests showed significant differences only for DY. Protein levels and mRNA expression were significantly increased for IL-8 for both periods of time. IL-6 production increased when fibroblasts were stimulated by KM. SDF-1α protein production and mRNA expression were not affected by any of the materials. There was a decrease in nitrate/nitrite levels only for KM. Although DY caused intense cell death and did not stimulate the production of the inflammatory mediators evaluated in this work, it is known that this event seems to be fundamental for the process of repair of the pulp tissue and formation of mineralized barrier. KM and VB increased production of proteins related to the inflammatory process, thus favoring tissue repair. Therefore, although these glass ionomer cements did not lead to large cell death, they should be used with caution.

Resumo Este estudo avalia in vitro a viabilidade e metabolismo celular, a liberação de óxido nítrico e a produção de duas quimiocinas e uma citocina por fibroblastos de polpa dentária humana em cultura (FPDH) em contato com dois cimentos de ionômero de vidro (Ketac Molar-KM e Vitrebond-VB), Single Bond (SB) e hidróxido de cálcio (Dycal-DY). As culturas de FPDH foram estabelecidas por meio de uma técnica de explante. As amostras foram preparadas em condições estéreis e em discos de 5 mm x 2 mm, obtidas de um molde pré-fabricado e colocadas em uma membrana permeável (Maxicell 24 W 0,4 µm) para evitar o contato direto com as células. A citotoxicidade foi avaliada pelo método de exclusão de Trypan Blue e pelo ensaio de MTT. A liberação de óxido nítrico no sobrenadante celular foi detectada pelo método Griess, enquanto fator 1 derivado do estroma (SDF-1α ou CXCL12), interleucina-8 (IL-8 ou CXCL8) and interleucina-6 (IL-6) foram detectados por ELISA. RT-qPCR foi empregada para análise de expressão gênica. As análises estatísticas foram realizadas por ANOVA a 1 critério, seguida pelo pós-teste de Tukey para os materiais independentes do tempo, e ANOVA a 2 critérios, seguida pelo teste de correção de Bonferroni para comparações entre materiais e tempo experimental (p<0,05). Os testes citotóxicos mostraram diferenças significativas apenas para DY. Os níveis da proteína e a expressão de RNAm para IL-8 aumentaram significativamente para ambos os tempos estudados. A produção de IL-6 aumentou quando os fibroblastos foram estimulados por KM. A produção da proteína e a expressão de RNAm para SDF-1α não foram afetadas por nenhum dos materiais. Houve uma diminuição nos níveis de nitrato/nitrito apenas para KM. Embora o DY tenha causado intensa morte celular e não tenha estimulado a produção dos mediadores inflamatórios avaliados neste trabalho, sabe-se que esse evento parece ser fundamental para o processo de reparo do tecido pulpar e formação de barreira mineralizada. Os cimentos de ionômero de vidro utilizados aumentaram a produção de proteínas relacionadas ao processo inflamatório, favorecendo a reparação tecidual e, portanto, esses materiais, embora não causem grande morte celular, devem ser utilizados com cautela.

Humans , Dental Pulp , Dental Pulp Capping , Fibroblasts
Braz. dent. j ; 31(3): 319-336, May-June 2020. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132301


Abstract This study aimed to quantitatively compare the difference in protein expression in the progression of pulp pathogenesis, as well as to describe the biological functions of proteins identified in pulp tissue. Samples were obtained from six patients treated at the Araçatuba School of Dentistry and were divided into three groups: normal pulp - from teeth extracted for orthodontic indication; inflamed pulp and necrotic pulp - from patients diagnosed with irreversible pulpitis and chronic apical periodontitis, respectively. After previous proteomic preparation, dental pulp samples were processed for label-free quantitative proteomic analysis in a nanoACQUITY UPLC-Xevo QTof MS system. The difference in expression between the groups was calculated using the Protein Lynx Global Service software using the Monte Carlo algorithm. A total of 465 human proteins were identified in all groups. The most expressed proteins in the inflamed pulp group in relation to the normal pulp group were hemoglobin, peroxiredoxins and immunoglobulins, whereas the less expressed were the tubulins. Expression levels of albumins, immunoglobulins and alpha-2-macroglobulin were higher in the necrotic pulp group than in the inflamed pulp group. As for the qualitative analysis, the most prevalent protein functions in the normal pulp group were metabolic and energetic pathways; in the inflamed pulp group: cellular communication and signal transduction; and regulation and repair of DNA/RNA, while in the necrotic pulp group proteins were associated with the immune response. Thus, proteomic analysis showed quantitative and qualitative differences in protein expression in different types of pulp conditions.

Resumo Este estudo teve como objetivo comparar quantitativamente a diferença da expressão de proteínas na progressão da patogênese pulpar, bem como descrever as funções biológicas das proteínas identificadas no tecido pulpar. As amostras foram obtidas de seis pacientes atendidos na Faculdade de Odontologia de Araçatuba e divididas em três grupos: polpa normal - dentes extraídos por indicação ortodôntica; polpa inflamada e polpa necrótica - pacientes diagnosticados com pulpite irreversível e periodontite apical crônica, respectivamente. Após o preparo proteômico prévio, as amostras de polpa dentária foram processadas para análise proteômica quantitativa livre de marcadores em um sistema nanoACQUITY UPLC-Xevo QTof MS. A diferença de expressão entre os grupos foi calculada usando o software Protein Lynx Global Service através do algoritmo de Monte Carlo. Um total de 465 proteínas humanas foram identificadas em todos os grupos. As proteínas mais expressas no grupo polpa inflamada em relação ao grupo polpa normal foram hemoglobinas, peroxirredoxinas e imunoglobulinas, enquanto as menos expressas foram as tubulinas. Os níveis de expressão de albuminas, imunoglobulinas e alfa-2-macroglobulina foram maiores no grupo polpa necrótica do que no grupo de polpa inflamada. Quanto à análise qualitativa, as funções proteicas mais prevalentes no grupo polpa normal foram vias metabólicas e energéticas; no grupo polpa inflamada: comunicação celular e transdução de sinal; e regulação e reparo de DNA / RNA, enquanto no grupo polpa necrótica as proteínas foram associadas à resposta imune. Assim, a análise proteômica mostrou diferenças quantitativas e qualitativas na expressão de proteínas em diferentes tipos de condições pulpares.

Humans , Pulpitis , Dental Pulp , Pilot Projects , Proteomics
Dental press j. orthod. (Impr.) ; 25(3): 85-92, May-June 2020. graf
Article in English | LILACS, BBO | ID: biblio-1133663


ABSTRACT Introduction: Stem cells obtained from the pulp of human deciduous teeth are highly proliferative and plastic multipotent cells, which makes them a relevant model of stem cells, applied in several biomedical areas, with different purposes. Objective: Based on a brief review of the literature, the present work intends to present from conceptual aspects about stem cells, classifications, potential (in vitro and in vivo) applications in dental practice, cell culture, cryopreservation and its importance, ethical and regulatory aspects, as well as the role of the dental surgeon as the endorser responsible for the entire clinical stage that involves the process of collecting stem cells obtained from dental pulps for cryopreservation, with a view to using them under appropriate conditions, in accordance with scientifically proven and justified good laboratory and clinical practices.

RESUMO Introdução: As células-tronco obtidas a partir da polpa de dentes decíduos humanos são células multipotentes altamente proliferativas e plásticas, o que as torna um modelo relevante de células-tronco, aplicado em diversas áreas biomédicas, com diferentes propósitos. Objetivo: A partir de uma breve revisão da literatura, o presente trabalho pretende apresentar desde aspectos conceituais acerca das células-tronco, classificações, potenciais aplicações (in vitro e in vivo) na prática odontológica, cultivo celular, criopreservação e sua importância, aspectos éticos e regulatórios, bem como o papel do cirurgião-dentista como homologador responsável por toda a etapa clínica que envolve o processo de coleta das células-tronco obtidas a partir de polpas dentais para criopreservação, com vistas ao uso em condições adequadas, em acordo com as boas práticas laboratoriais e clínicas cientificamente comprovadas e justificadas.

Humans , Adult , Mesenchymal Stem Cells , Stem Cells , Tooth, Deciduous , Dental Pulp , Dentistry
Dent. press endod ; 10(1): 54-61, Jan-Apr2020. Tab, Ilus
Article in English | LILACS | ID: biblio-1344238


Pacientes submetidos à clareação dentária relatam sensibilidade pós-operatória relacionada ao peróxido de hidrogênio (H2 O2 ) que penetra no tecido pulpar. Objetivo: Avaliar o efeito anti-inflamatório do ibuprofeno, Otosporin® e gel de curcumina na polpa dentária de ratos após procedimento clareador. Métodos: Cinquenta ratos foram divididos em GC ­ controle (gel placebo); CLA ­ clareação (H2 O2 35%, 30 minutos); CLA-I ­ clareação e administração oral de ibuprofeno (duas vezes a cada 12 horas, 2 dias sucessivos); CLA-O ­ clareação seguida da aplicação de Otosporin® nas superfícies dos molares (10 minutos); e CLA-C ­ sessão clareadora seguida do gel de curcumina (10 minutos). Após dois dias, os ratos foram mortos para análise histológica e testes estatísticos foram realizados(p<0,05). Resultados: CLA, CLA-I e CLA-C apresentaram inflamação severa ou necrose no terço oclusal da polpa coronária (p>0,05); CLA-O apresentou inflamação leve e foi semelhante ao GC (p>0,05) e dife- rente dos outros grupos (p<0,05). No terço médio, o grupo CLA-O apresentou menor infiltrado inflamatório e permaneceu diferente do grupo CLA (p<0,05); CLA, CLA-I e CLA-C foram semelhantes (p>0,05). No terço cervical, CLA, CLA-I e CLA-C tiveram redução da inflamação, sem diferença entre os grupos clareados (p>0,05). Conclusões: O Otosporin® pode reduzir a inflamação na polpa após clareação dentária; esse resultado não foi observado utilizando ibuprofeno ou gel de curcumina. Portanto, esse estudo mostra uma nova possibilidade de pós-tratamento em dentes clareados por meio do uso de Otosporin®, que minimiza a inflamação gerada ao tecido pulpar pelo gel clareador. Consequentemente, poderá haver redução da sensibilidade pós-operatória (AU).

Introduction: Patients undergoing dental bleaching relate to postoperative sensitivity, that is linked to hydrogen peroxide (H2 O2 ) penetrating on the dental pulp. This study evaluated the anti-inflammatory effect of ibuprofen, Otosporin®, and curcumin gel on the pulp of the rats' teeth after bleaching. Methods: Fifty rats were divided into CG: controlplacebo gel; BLE: bleached (35% H2O2, 30 minutes); BLE-I: bleached and ibuprofen oral administration (twice every 12 hours in 2 successive days); BLE-O: bleached followed by Otosporin® application in the molar surfaces (10 minutes); and BLE-C: bleaching session followed curcumin gel (10 minutes). After two days, the rats were killed for histological analysis. Statistical tests were performed (P<.05). Results: BLE, BLE-I, and BLE-C had severe inflammation or necrosis in the occlusal third of coronal pulp (P>.05); BLE-O had mild inflammation and was similar from CG (P>.05) and different from other groups (P<.05). In the middle third, BLE-O group had lower inflammatory infiltration and remained different from BLE group (P<.05); BLE, BLE-I, and BLE-C were similar (P>.05). In the cervical third, BLE, BLE-I, and BLE-C had a reduction of inflammation, without difference between bleached groups (P>.05). Conclusions: Otosporin® can reduce the inflammation in the pulp after dental bleaching; this result was not observed using ibuprofen or curcumin gel. Therefore, this study shows a new teeth bleaching posttreatment possibility using Otosporin®, which minimizes the inflammation generated to the pulp tissue by the bleaching gel. This could consequently minimize the postoperative sensitivity (AU).

Animals , Rats , Tooth Bleaching , Dental Pulp , Hydrogen Peroxide , Anti-Inflammatory Agents , Curcumin
Int. j. morphol ; 38(2): 322-327, abr. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1056442


La estimación de edad compone un aspecto importante en investigaciones forenses. Diferentes métodos se han descrito en odontología forense basadas en la correlación entre la edad y estructuras dentales. Cameriere et al. proponen un método cuantitativo para estimación de edad en adultos, a partir de la evaluación de la relación del área pulpa/diente, en base a la aposición de dentina secundaria. El objetivo del estudio fue desarrollar modelos de regresión para la estimación de edad dental mediante la relación área pulpa/diente en caninos inferiores en una muestra Chilena. Se analizaron 212 radiografías periapicales digitales (RPD) (86 hombres y 126 mujeres) de caninos mandibulares mediante el programa Image J para establecer el área de la pulpa y el diente. Se registraron los datos de sexo y edad de las RPD seleccionadas en forma ciega. Fueron desarrollados modelos de regresión lineal simples para la estimación de edad. El coeficiente de determinación para R33 fue 27,8 % y de 29,6 % para R44, con un error absoluto medio de 11,02 años y 10,37 años respectivamente. El análisis de ANOVA no mostró diferencias estadísticamente significativas para las relaciones área pulpa/diente de caninos según sexo (p> 0,05). Según los resultados obtenidos, la metodología propuesta por Cameriere et al. es fiable para estimar la edad dental mediante la relación área pulpa/diente en adultos. Sin embargo, en los modelos de regresión desarrollados para la población Chilena, se puede afirmar que el ajuste indicado por los coeficientes de determinación muestran incerteza entre las variables área pulpa/diente y edad cronológica en caninos inferiores, por lo tanto se sugiere considerar otros métodos adicionales para estimar edad en esta población.

Age estimation is an important aspect In forensic investigations. Different methods in forensic odontology based on the correlation between age estimation in adults, from the analysis of the pulp/tooth area, based on the apposition of secondary dentine. The aim of the study was to develop regression models for the dental age estimation by the relation pulp/tooth area, in lower canines in a Chilean sample, using digital peri-apical radiographs (DPR) applying Cameriere's method. We analyzed 212 DPR (86 males and 126 females) mandibular canines through Image J program to measure the pulp/tooth area. Age and sex information was obtained of the DPR's blindly selected. We developed simple linear regression models for age estimation. The coefficient of determination to R33 was R2 age and dental structures have been described. Cameriere et al. proposed a quantitative method for 27.8 % and R2 29.6 % to R44, with a mean absolute error of 11.02 years, to R33 and 10.37 years to R44. ANOVA analysis showed no statistically significant differences for the pulp/tooth relation area of canines according to sex (p> 0.05). According to the results, the Cameriere's et al., method is reliable for dental age estimation according to pulp/tooth ratio in adults. However, in the regression models developed for Chilean population, it can be stated that the adjustment indicated by the coefficients of determination, show uncertainty between the pulp / tooth area and chronological age in lower canines, therefore it is suggested to use additional estimation methods for age in this population.

Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Tooth/anatomy & histology , Age Determination by Teeth/methods , Radiography, Dental, Digital/methods , Dental Pulp/anatomy & histology , Tooth/diagnostic imaging , Logistic Models , Chile , Analysis of Variance , Dental Pulp/diagnostic imaging , Age and Sex Distribution , Forensic Dentistry
Article in English | WPRIM | ID: wpr-811428


OBJECTIVES: This study investigated the indirect effect of calcium-enriched mixture (CEM) cement and mineral trioxide aggregate (MTA), as 2 calcium silicate-based hydraulic cements, on human dental pulp stem cells (hDPSCs) through different dentin thicknesses.MATERIALS AND METHODS: Two-chamber setups were designed to simulate indirect pulp capping (IPC). Human molars were sectioned to obtain 0.1-, 0.3-, and 0.5-mm-thick dentin discs, which were placed between the 2 chambers to simulate an IPC procedure. Then, MTA and CEM were applied on one side of the discs, while hDPSCs were cultured on the other side. After 2 weeks of incubation, the cells were removed, and cell proliferation, morphology, and attachment to the discs were evaluated under scanning electron microscopy (SEM). Energy-dispersive X-ray (EDXA) spectroscopy was performed for elemental analysis. Alkaline phosphatase (ALP) activity was assessed quantitatively. The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests.RESULTS: SEM micrographs revealed elongated cells, collagen fibers, and calcified nucleations in all samples. EDXA verified that the calcified nucleations consisted of calcium phosphate. The largest calcifications were seen in the 0.1-mm-thick dentin subgroups. There was no significant difference in ALP activity across the CEM subgroups; however, ALP activity was significantly lower in the 0.1-mm-thick dentin subgroup than in the other MTA subgroups (p < 0.05).CONCLUSIONS: The employed capping biomaterials exerted biological activity on hDPSCs, as shown by cell proliferation, morphology, and attachment and calcific precipitations, through 0.1- to 0.5-mm-thick layers of dentin. In IPC, the bioactivity of these endodontic biomaterials is probably beneficial.

Alkaline Phosphatase , Biocompatible Materials , Calcium , Cell Proliferation , Collagen , Dental Pulp Capping , Dental Pulp , Dentin , Endodontics , Humans , Microscopy, Electron, Scanning , Miners , Molar , Pemetrexed , Spectrum Analysis , Stem Cells
Article in Chinese | WPRIM | ID: wpr-827539


Tubular dentin is of great significance in the process of tooth tissue and tooth regeneration, because it is not only the structural feature of primary dentin, but also can affect the tooth sensory function, affect the differentiation of dental pulp cells and provide strong mechanical support for teeth. Scaffold is one of the three elements of tissue engineering dentin regeneration. Most experiments on dentin regeneration involve the study of the microstructure and mechanical properties of the scaffold. The microstructure and mechanical characteristics of scaffold materials have important effects on the differentiation and adhesion of odontoblast, it can directly affect the tissue structure of regenerated dentin.

Cell Differentiation , Dental Pulp , Dentin , Odontoblasts , Regeneration , Tissue Engineering , Tissue Scaffolds