The response of Mesenchymal Stem Cells to endodontic materials

Abstract An endodontic material must be minimally harmful to stem cells since they are essential, thanks to their capacity for cell proliferation, self-renewal, and differentiation. For this reason, in this in vitro study, the cell viability and the expression of genes involved in cell plasticity and differentiation were investigated in stem cells recovered from human dental pulp (hDPSCs) that were in contact with four endodontic materials (Endofill, MTA, Pulp Canal Sealer, and Sealer 26). The viability of HDPSCs was assessed by MTT and trypan blue exclusion assays. PCR evaluated cellular plasticity by determining the CD34, CD45, Nestin, CD105, Nanog, and OCT4 expressions. The effect on cell differentiation was determined by RT-PCR expression of the RUNX2, ALP, OC/BGLAP, and DMP1 genes. The data were analyzed using ANOVA with Bonferroni correction (p <0.05). Pulp Canal Sealer and Endofill decreased cell viability after 48 hours (p <0.001). MTA and Sealer 26 did not disrupt cell viability (p> 0.05). When cultivated in the presence of MTA and Sealer 26, hDPSCs expressed Nestin, CD105, NANOG, and OCT-4 and did not express CD34 and CD45. MTA and Sealer 26 interfered with DMP1, OC/BGLAP and RUNX2 expressions (p <0.05) but did not change ALP gene expression (p> 0.05). MTA and Sealer 26 showed biological compatibility in the presence of hDPSCs.

Resumo Um material endodôntico deve ser minimamente prejudicial às células-tronco, uma vez que essas células são extremamente importantes, devido à sua capacidade de proliferação, autorrenovação e diferenciação celular. Por esse motivo, a viabilidade celular e a expressão de genes envolvidos na plasticidade e diferenciação celular foram investigadas em células-tronco recuperadas de polpa dentária humana (HDPSCs) que estiveram em contato com quatro materiais endodônticos (Endofill, MTA, Pulp Canal Sealer e Sealer 26). A viabilidade das HDPSCs foi avaliada pelos ensaios MTT e de exclusão de azul de tripano. A plasticidade celular foi avaliada pela determinação das expressões dos genes CD34, CD45, Nestin, CD105, Nanog e OCT4 por PCR. O efeito na diferenciação celular foi determinado pela expressão dos genes RUNX2, ALP, OC/BGLAP e DMP1 por RT-PCR. Os dados foram analisados por ANOVA com correção de Bonferroni (p <0,05). Em comparação com o controle, Pulp Canal Sealer e Endofill diminuíram a viabilidade celular após 48 horas (p <0,001). MTA e Sealer 26 não interromperam a viabilidade celular (p> 0,05). Quando cultivado na presença de MTA e Sealer 26, as HDPSCs expressaram Nestin, CD105, NANOG e OCT-4 e não expressaram CD34 e CD45. MTA e Sealer 26 interferiram nas expressões de DMP1, OC / BGLAP e RUNX2 (p <0,05), mas não alteraram a expressão do gene ALP (p> 0,05). Sendo assim, MTA e Sealer 26 demonstraram compatibilidade biológica na presença de HDPSCs.

Experimental furcal perforation treated with mineral trioxide aggregate plus selenium: immune response

Abstract The aim of this study is to evaluate the expression of cytokines in response to mineral trioxide aggregate (MTA) plus selenium in germ-free mice with experimental furcal perforation. The first left maxillary molar was opened, and the furcal area was perforated and treated with post-MTA-Se (experimental group). The same surgical intervention was performed for the maxillary right first molar, which was treated with MTA (control group). Fifteen mice were sacrificed 7, 14, and 21 days after furcal perforation, and periapical tissue samples were collected. The mRNA expression levels of the cytokines TGF-β, TNF-α, IFN-γ, HPRT, IL-10, IL-4, RANK, RANKL, IL-1, and IL-17 were assessed by using real-time polymerase chain reaction. In the experimental group, at 21-days post-MTA-Se sealing, the mRNA levels of TNF-α and IL-10 were upregulated compared with those in the control group (p < 0.05). Futher assessment revealed basal mRNA expression levels of IL-1α, IFN-γ, RANK, RANKL, IL-17A, IL-4, and TGF-β, over long experimental times, in both the experimental and control groups (p > 0.05). In conclusion, MTA+Se sealing favoured increased expression of IL-10 and TNF-α at later time points (day 21).

Epidemiological analysis and need for endodontic treatment among the indigenous Sateré-Mawé and Tikuna

Abstract: The objective of this study was to analyze the epidemiological profile of oral health of Sateré-Mawé indigenous people living in Barreirinha, Amazonas (AM), Brazil, and the Tikuna indigenous people living in the urban area of Manaus (AM), in addition to characterizing the need for endodontic treatment between the two ethnic groups. A total of 138 individuals participated in the study, of whom 98 were Tikuna and 40 were Sateré-Mawé; they were distributed in age groups ranging from seven to 75 years. A very high prevalence of caries was observed in both ethnic groups. For the Sateré-Mawé in the 7-12 age group, the decayed, missing, and filled teeth (DMFT) index presented a mean value of 3.17. Comparing the DMFT index and the need for endodontic treatment in each of the ethnicities, these variables were found to be correlated, because as the DMFT index increases, the chances of needing endodontic treatment increase. The Sateré-Mawé presented a higher prevalence of need for endodontic treatment compared to the Tikuna. The association of comorbidities and the need for endodontic treatment were demonstrated only in the Tikuna, and there was only a correlation of this necessity with the presence of diabetes mellitus (DM) in one case. The need to expand access to oral health in these communities is emphasized, taking into account geographical access and technological, environmental, linguistic, and cultural barriers.

The need for endodontic treatment and systemic characteristics of hematopoietic stem cell transplantation patients

Abstract The aim of this study is to investigate the relationship between the epidemiological and clinical profiles of patients before and after hematopoietic stem cell transplantation (HSCT) and the need for endodontic treatment. The subjects included 188 individuals enrolled in the dental care program for transplanted patients of the School of Dentistry, Federal University of Minas Gerais (Faculdade de Odontologia da Universidade Federal de Minas Gerais, FO-UFMG) from March 2011 through March 2016. The patients were subjected to an HSCT conditioning dental regimen based on a thorough clinical and radiographic evaluation. Intraoral periapical and bite-wing X-rays were obtained, and after evaluation, specific dental treatment was planned and performed. The following demographic and clinical data were collected from the patients' medical records: age, gender, transplantation stage, primary disease, transplant type, medication used, complete blood count at the time of visit, and need for endodontic treatment. The Kolmogorov-Smirnov and the chi-square tests were used. Leukemia (31.3%) and multiple myeloma (17.9%) were the most prevalent primary diseases. Most patients were subjected to allogeneic-related transplantation (83.6%). Most patients exhibited platelet counts and hemoglobin concentrations below the reference values in the pre-transplantation stage, while the neutrophil and platelet counts and the hemoglobin levels were within the reference ranges in the post-transplantation stage. The proportions of individuals requiring endodontic treatment were similar between the pre- and post-transplantation groups: 24.3% and 24.7%, respectively. The systemic conditions of the patients referred for dental treatment were compromised.

Murine Experimental Root Canal Infection: Cytokine Expression in Response to F. nucleatum and E. faecalis

Abstract The aim of this study was to evaluate the gene expression of proinflammatory (RANKL, TNF-a and IFN-g) and regulatory (TGF-b and IL-10) cytokines as reaction to experimental infection by mono or bi-association of Fusobacterium nucleatum (ATCC 10953) and Enterococcus faecalis (ATCC 19433). F. nucleatum and E. faecalis, either in mono- or bi-association were inoculated into the root canal system (RCS) of Balb/c mice. Animals were sacrificed at 10 and 20 days after infection and periapical tissues surrounding the root were collected. The mRNA expression of the cytokines RANKL, TNF-a, IFN- g, TGF-b and IL-10 was assessed using real-time PCR. The Kruskal-Wallis test was used for statistical analysis. F. nucleatum mono-infection induced high expression of RANKL and TNF-a, while its modulation was due to IL-10. High expression of IFN-g at day 20 was up-regulated by E. faecalis and RANKL; TNF-a was up-regulated by an independent mechanism via IL-10 and TGF-b. Bi-association (F. nucleatum and E. faecalis) stimulated high expression of RANKL, TNF-a and IFN-g, which seemed to be modulated by TGF-b 20 days later. The gene expression of proinflammatory cytokines was more prominent in the earlier periods of the experimental periapical infection, which concomitantly decreased in the later period. This expression may be regulated by IL-10 and TGF-b in an infection-specific condition

Resumo O objetivo deste trabalho foi avaliar a expressão gênica de citocinas pró-inflamatórias (RANKL, TNF-a e IFN-g) e regulatórias (TGF-b e IL-10) em resposta à infecção experimental por Fusobacterium nucleatum (ATCC 10953) e Enterococcus faecalis (ATCC 19433) como mono-infecção ou em bi-associação. F. nucleatum e E. faecalis foram inoculados no sistema de canais radiculares de camundongos Balb/c, tanto isoladas como em bi-associação. Os animais foram sacrificados em 10 e 20 dias após a infecção, e os tecidos periapicais foram coletados. As expressões do mRNA das citocinas RANKL, TNF-a, IFN-g, TGF-b e IL-10 foram analisadas por meio do real-time PCR. O teste de Kruskal-Wallis foi utilizado para análise estatística. A mono-infecção com F. nucleatum induziu alta expressão de RANKL e TNF-a, enquanto sua modulação ocorreu devido à IL-10. A alta expressão de IFN-g no dia 20 foi regulada positivamente por E. faecalis e RANKL; TNF-a foi regulada positivamente por um mecanismo independente via IL-10 e TGF-b. A bi-associação (F. nucleatum e E. faecalis) estimulou uma alta expressão de RANKL, TNF-a e IFN-g, que parece ser modulada por TGF-b após 20 dias. A expressão gênica de citocinas pró-inflamatórias foi mais proeminente nos estágios iniciais da infecção periapical experimental, com concomitante redução no período tardio. Este fenômeno pode ser regulado por IL-10 e TGF-b em uma condição de infecção específica.

Sickle cell anemia in Brazil: personal, medical and endodontic patterns

Abstract Sickle cell anemia (SCA) is the most prevalent genetic disease worldwide. Recurrent vaso-occlusive infarcts predispose SCA patients to infections, which are the primary causes of morbidly and mortality. This study aimed to evaluate the relationship between SCA and endodontic diseases. Personal information, medical data (hematological indices, virologic testing, blood transfusions, medications received, splenectomy) and information on the need for endodontic treatment were obtained from SCA patients who were registered and followed up by the Fundação Hemominas, Minas Gerais, Brazil.These data were compared with the need for root canal treatment in SCA patients. One hundred eight patients comprised the studied population, and the rate of the need for endodontic therapy was 10.2%. Among the medical data, a significant difference was observed for eosinophil (p = 0.045) counts and atypical lymphocyte counts (p = 0.036) when the groups (with and without the need for endodontic treatment) were compared. Statistical relevance was observed when comparing the patients with and without the need for root canal therapy concerned eosinophil counts and atypical lymphocyte counts. The differences in statistical medical data, observed between the groups suggest that both parameters are naturally connected to the stimulation of the immune system that can occur in the presence of root canal infections and that can be harmful to SCA individuals.

Experimental Furcal Perforation Treated with MTA: Analysis of the Cytokine Expression

The aim of this study was to evaluate mineral trioxide aggregate (MTA)-induced cytokine expression in mice after experimental furcal perforation. BALB/c mice (n=5) were subjected to induced furcal drilling of the maxillary first molar followed by MTA sealing in the left side (experimental group) and paraffin sealing in the right side (control group). Animals were euthanized at 7, 14 and 21 days after sealing the perforations. The expression levels of the IFN-γ, TNF-α, IL-10, IL-4, TGF-β and RANKL genes were investigated by real time polymerase chain reaction (PCR) in the teeth and surrounding tissues. In the experimental groups, after the 7th day, there was a down-regulation of the mRNA levels of TNF-α and IL-4 compared to the 14th day (p<0.05). In these groups, the mRNA levels of RANKL, IFN-γ and TNF-α were statistically higher after 14 days compared to 21 days post-MTA sealing (p<0.05). The level of IL-10 mRNA was increased at the 21st day (p<0.05). The mRNA expression of TGF-β did not exhibit any statistically relevant results. There was a statistical down-regulation of IL-4 gene expressions when control and experimental groups were compared at days 7 and 21. In conclusion, MTA sealing favored the expression of pro-inflammatory cytokines in the intermediate phase of the immuno-inflammatory response (14th day). The reduction of these cytokines in later phase of the response was probably due to immunoregulation by IL-10.

O objetivo desse artigo foi avaliar a expressão das citocinas induzidas por MTA após a perfuração experimental de furca, em camundongos. Camundongos Balb/c (n=5) foram submetidos à perfuração induzida da furca do primeiro molar superior, seguido pelo selamento da mesma com MTA no lado esquerdo (grupo experimental) e com parafina no lado direito (grupo controle). Os animais foram sacrificados 7, 14 e 21 dias após o tratamento da perfuração. A expressão gênica dos níveis de IFN-γ, TNF- α, IL-10, IL-4, TGF- β e RANKL foram investigadas por PCR em tempo real nos dentes e tecidos adjacentes. No grupo experimental, após 7 dias, houve uma diminuição da expressão dos níveis de TNF- α e IL-4 comparados ao 14° dia (p<0,05). Nesses mesmos grupos, os níveis de mRNA de RANKL, IFN- γ e TNF- α foram estatisticamente maiores após 14 dias comparados a 21 dias após o tratamento com MTA (p<0,05). Os níveis de IL-10 estavam aumentados no 21^o dia (p<0,05). A expressão de mRNA do TGF- β não apresentou alteração estatisticamente relevante. Houve uma redução estatística da expressão gênica da IL-4 quando os grupos controle e experimental foram comparados nos dias 7 e 21. Em conclusão, o selamento com MTA favoreceu a expressão de citocinas pró-inflamatórias na fase intermediária da resposta imuno-inflamatória (14^o dia). A redução dessas citocinas, na fase tardia da resposta, ocorreu provavelmente devido à imunoregulação da expressão de IL-10.

Low and high-dose intradermal infection with Leishmania majorand Leishmania amazonensis in C57BL/6 mice

A model of skin infection with Leishmania amazonensiswith low doses of parasites is compared to infection with high doses of L. amazonensis and low and high doses of Leishmania major. C57BL/6 mice were infected with 10³ or 10(6) parasites in the ear and the outcome of infection was assessed. The appearance of lesions in mice infected with 10³ parasites was delayed compared to mice infected with 10(6) Leishmania and parasites were detectable at the infection site before lesions became apparent. Mice infected with L. amazonensisdisplayed persistent lesions, whereas infection with L. major spontaneously healed in all groups, although lymphocytes persisted at the site of infection after healing. Macrophages persisted only in L. amazonensis-infected mice. High-dose L. amazonensis-infected mice produced lower levels of IFN-γ and TNF than mice infected with L. major. No correlation between the persistence of parasites and IL-10 levels and the production of nitric oxide or urea by macrophages was found. We conclude that infection with low doses of L. amazonensisin the dermis changes the course of infection by delaying the appearance of lesions. However, low-dose infection does not change the outcomes of susceptibility and cytokine production described for subcutaneous infection with high numbers of parasites.

Histopathology of Leishmania major infection: revisiting L. major histopathology in the ear dermis infection model

We describe the relationship between lesion outcome and histopathological hallmarks in susceptible (BALB/c) and resistant (C57BL/6 and IL-4-deficient BALB/c) mouse strains over the course of a 12-week-infection with Leishmania major in the ear. The infiltration of mononuclear cells and polymorphonuclear cells occurred within 6 h and mononuclear cells predominated one week post-infection. Permissive intracellular growth of the pathogen was associated with non-healing lesions. In contrast, tissue damage and clearance of the parasite was observed in healing lesions and was associated with inducible nitric oxide synthase expression. The identification of the structural components of tissue reaction to the parasite in this study furthers our understanding of subjacent immune effector mechanisms.

Uso de probióticos na prevenção e tratamento de infecções e inflamações gastrintestinais / The use of probiotics in the prevention and treatment of gastrointestinal infection and inflammation

A microbiota indígena associada ao trato digestivo humano é um ecossistema extremamente complexo e pouco conhecido. A sua instalação no recém-nascido e sua manutenção no adulto têm papel importante na maturação da biologia e proteção contra infecção do hospedeiro, respectivamente. Contudo, ambas podem ser perturbadas por diversos fatores, levando a uma redução dos benefícios que a atuação da microbiota pode oferecer em condições normais. Esses distúrbios podem ser compensados pelo uso de probióticos, definidos como microorganismos vivos que quando aplicados em quantidade adequada podem oferecer benefícios para o hospedeiro. Bactérias produtoras de ácido láctico (Lactobacillus, bifidobacterium) e levedura (Saccharomyces boulardii) são entre os microorganismos mais estudados ou já usados como probióticos. Para certas aplicações, a soma de ensaios clínicos já foi suficiente para efetuar meta-análises que mostraram efeito positivos dos probióticos, como na profilaxia de diarréia aguda e da diarréia associada ao uso de antibióticos (em particular no caso de infecção pelo clostridium difficile) e como suplemento na terapia contra Helicobacter pylori. Mais recentemente, efeitos positivos foram também sugeridos no caso de doenças inflamatórias intestinais. Contudo, todos os autores concordam com a necessidade de mais ensaios clínicos e biológicos, em particular para esclarecer os mecanismo de ação de probióticos e os fatores do ecossistema digestivo que podem influir sobre essa atuação. Tais esclarecimento devem permitir otimizar o uso dos probióticos, regularizando resultados que hoje são algumas vezes flutuantes.

Leishmania braziliensis: partial control of experimental infection by interleukin-12 p40 deficient mice

Resistance to infection by Leishmania major has been associated with the development of a Th1 type response that is dependent on the presence of interleukin 12 (IL-12). In this work the involvement of this cytokine in the response to infection by L. braziliensis, a less virulent species in the mouse model, was evaluated. Our results show that while interferon (IFN-gamma) deficient (-/-) mice inoculated L. braziliensis develop severe uncontrolled lesions, chronic lesions that remained under control up to 12 weeks of infection were observed in IL-12p40 -/- mice. IL 12p40 -/- mice had fewer parasites in their lesions than IFN-gamma-/- mice. Lymph node cells from IL-12p40 -/- were capable of producing low but consistent levels of IFN-gamma suggestive of its involvement in parasite control. Furthermore, as opposed to previous reports on L. major-infected animals, no switch to a Th2 response was observed in IL-12p40 -/- infected with L. braziliensis.

Probioticos / Probiotics

A microbiota normal exerce papel importante na proteção contra agentes infecciosos por mecanimos ecológicos e imunológicos, além de contribuir para a nutrição do hospedeiro. Distúrbios na microbiota acarretam prejuízos desses efeitos. A ingestão de probióticos pode prevenir os efeitos dos distúrbios da microbiota. Os probióticos estudados e utilizados em humanos são bactérias láticas (Lactobacillus, Bifidobacterium, Streptococcus e Enterococcus) e leveduras (Saccharomyces boulardii). Vários mecanismos de ação, obtidos a partir de estudos experimentais, já foram propostos para os probióticos: a proteção ecológica, seja pela prevenção da multiplicação dos patógenos ou pela inibição da ação patogênica e modulação do sistema imune, por ativação do sistema fagocitário, produção de imunoglobulinas e citocinas. Alguns ensaios clínicos controlados demonstram que os probióticos podem ser utilizados com sucesso na prevenção ou tratamento de distúrbios entéricos. Concluindo, os probióticos podem ser utilizados na compensação de uma redução prevista ou instalada das funções da microbiota digestiva.

The normal microbiota has an important role on the protection against infectious agents, acting by ecological or immunological mechanisms. In addition, it contributes to the host's nutrition. Microbiota instabilities lead to the loss of these effects. Ingestion of probiotics may prevent the effects of microbiota instabilities. Most probiotics under study and utilized in humans are lactic bacteria (Lactobacillus, Bifidobacterium, Streptococcus e Enterococcus) and yeasts (Saccharomyces boulardii). Several mechanisms of action for probiotics are proposed, mostly based on experimental studies: ecological protection, either by prevention of multiplication of pathogens or inhibition of pathogenic action, and immune modulation by activation of phagocytes, production of immunoglobulins and cytokines. Controlled clinical assays show that probiotics may be successful in preventing or treating enteric diseases. In conclusion, probiotics may be used to compensate, either profilactically or therapeutically, a reduction of the function of the normal gut microbiota.

Protection by Lactobacillus acidophilus UFV-H2B20 against experimental oral infection with Salmonella enterica subsp. Enterica Ser. Typhimurium in gnotobiotic and conventional mice

The ability of Lactobacillus acidophilus UFV-H2B20 to antagonize Salmonella enterica subsp. enterica ser. Typhimurium and to reduce the pathological consequences for the host was determining using conventional and gnotobiotic animals. Conventional NIH mice received daily by gavage a 0.1 ml suspension containing about 10(8) cfu L. acidophilus UFV-H2B20 and germfree animals received a single 0.1 ml dose. The gnotobiotic and conventional groups were infected orally with 10(2) and 10(5) cfu of S. Typhimurium, respectively, 7 days after the beginning of treatment. Control groups were treated with sterile saline instead of Lactobacillus. Survival data showed a protective effect against the pathogenic bacteria in both conventional and gnotobiotic Lactobacillus-treated mice. L. acidophilus UFV-H2B20 colonized the digestive tract of gnotobiotic mice and the number of viable cells ranged from 10(9) to 10(10) cfu/g of faeces. In both experimental and control gnotobiotic animals, S. Typhimurium became rapidly established at a level ranging from 108 to 1010 cfu/g of faeces and remained at high levels until the animals died or were sacrificed. In conclusion, the previous treatment of mice with L. acidophilus UFV-H2B20 protects the animals against the experimental infection with S. Typhimurium but this protection was not due to the reduction of the pathogenic populations in the intestines.(au)

Influence of microbiota in experimental cutaneous leishmaniasis in Swiss mice

Neste trabalho, estudamos a infeccao experimental de camundongos Suicos/NIH com Leishmania major em comparacao com camundongos isogenicos C57BL/6 e BALB/c que sao resistentes e suceptiveis a esta infeccao, respectivamente. Camundongos Suicos mostraram lesoes que se curaram espontaneamente e se restringiram ao local de inoculacao. Celulas linfoides derivadas destes animais desenvolveram uma resposta do tipo Th1, caracterizada pela producao de niveis altos de IFN-gama e niveis baixos de IL-4. Com o objetivo de estudar a importancia da microbiota no desenvolvimento da leishmaniose cutanea neste modelo, camundongos Suicos sem germes foram infectados na pata com promastigotas de L. major, convencionalizados apos 3 semanas de infeccao e as lesoes comparadas com as observadas em camundongos convencionais...

Schistosomicidal activity of alkylaminooctanethiosulfuric acids

The schistosomicidal activity of a new series of alkylaminooctanethiosulfuric acids was studied in white Swiss mice infected with the L.E. strain of Schistosoma mansoni (Belo Horizonte, MG, Brazil). In a preliminary screening of six compounds, two derivatives - 2-[(1-methylpropyl)amino]-1-octanethiosulfuric acid and 2-[(1-methylethyl)-amino]-1-octanethiosulfuric acid - given orally in doses of 300 mg/kg/day for five consecutive days, caused interruption of the oviposition and the hepatic shift of more than 90 of the worms. Both compounds caused a significant reduction in worm burden and, interestingly, the female schistosomes were more susceptible. With the therapeutic schedule of two doses of 800 mg/kg over a 20 day interval, the death of almost all the females and about 50 of the males was observed. Female worms recovered from treated mice showed scattered vitteline glands. Results of in vitro experiments against different developmental stages of the parasite revealed the induction of paralysis and damage to the tegument membrane. The drugs presented no toxic effects on the animals.