Systemic lithium chloride administration improves tooth extraction wound healing in estrogen-deficient rats

Abstract The purpose of this investigation was to evaluate the effects of lithium chloride (LiCl) on the socket healing of estrogen-deficient rats. Seventy-two rats were allocated into one of the following groups: Control, Ovariectomy and LiCl (150 mg/kg/2 every other day orally) + Ovariectomy. Animals received LiCl or water from the 14th day post-ovariectomy, until the completion of the experiment. On the 21st day after ovariectomy, the first molars were extracted. Rats were euthanized on the 10th, 20th and 30th days following extractions. Bone healing (BH), TRAP positive cells and immunohistochemical staining for OPG, RANKL, BSP, OPN and OCN were evaluated. The Ovariectomy group presented decreased BH compared to the LiCl group at 10 days, and the lowest BH at 20 days (p<0.05). At 30 days, the Ovariectomy and LiCl-groups presented lower BH than that of the Control (p<0.05). The number of TRAP-stained cells was the lowest in the LiCl group at 20 days and the highest in the Ovariectomy group at 30 days (p<0.05). At 10 days of healing, the LiCl group demonstrated stronger staining for all bone markers when compared to the other groups, while the Ovariectomy group presented higher RANKL expression than that of the Control (p<0.05). LiCl enhanced bone healing in rats with estrogen deficiency, particularly in the initial healing phases. However, as data on the effects of lithium chloride on bone tissue are still preliminary, more studies related to its toxicity and protocol of administration are necessary before its application in clinical practice.

Resumo O objetivo deste estudo foi avaliar os efeitos do Cloreto de Lítio (ClLi) na cicatrização de alvéolos de ratas deficientes em estrogênio. Setenta e duas ratas foram alocadas em um dos seguintes grupos: Controle, Ovariectomia e Cloreto de Lítio (150mg/kg/ oralmente a cada 2 dias) + ovarectomia. Os animais receberam ClLi ou água a partir do 14º dia pós-ovariectomia, até a conclusão do experimento. No 21º dia após a ovariectomia, os primeiros molares foram extraídos. As ratas foram sacrificadas nos dias 10, 20 e 30 após extrações. Foram avaliadas a cicatrização óssea (BH), células positivas para TRAP e coloração imuno-histoquímica para OPG, RANKL, BSP, OPN e OCN. O grupo Ovariectomia apresentou BH diminuída em comparação ao grupo LiCl aos 10 dias e a menor BH aos 20 dias (p<0,05). Aos 30 dias, os grupos Ovariectomia e LiCl apresentaram menor BH do que o Controle (p<0,05). O número de células positivas para TRAP foi menor no grupo ClLi em 20 dias e o maior no grupo Ovariectomia em 30 dias (p<0,05). Aos 10 dias de cicatrização, o grupo ClLi demonstrou imunomarcação mais intensa em todos os marcadores testados quando comparado aos outros grupos, enquanto o grupo Ovariectomia apresentou maior expressão de RANKL do que a do controle (p<0,05). O ClLi melhorou a cicatrização óssea em ratos com deficiência de estrogênio, particularmente nas fases iniciais do reparo. No entanto, como os dados sobre os efeitos do cloreto de lítio no tecido ósseo ainda são preliminares, mais estudos relacionados à sua toxicidade e protocolo de administração são necessários antes de sua aplicação na prática clínica.

Microbiological Seal of Two Types of Tapered Implant Connections

Abstract Tapered implant connections have gained wide popularity for being more resistant to fatigue and for promoting a better seal against bacterial infiltration than conventional connections. The aim of this study was to evaluate the bacterial seal at the implant-abutment interface using two Morse taper implant models, by in vitro microbiological analysis. Eleven non-indexed and 11 indexed abutments were selected and connected to their respective implants with a 20 N torque, according to manufacturer's recommendation. Microbiological analysis was carried out using colonies of Escherichia coli transported directly from a culture dish to the prosthetic component. For control, one non-contaminated abutment-implant set from each group (negative control) and one contaminated implant with no abutment (positive control) were used. The specimens were immersed in BHI broth and maintained in an incubator at 37 °C for 14 days to assess the development of bacterial contamination. The results revealed that 36.4% (n=4) of the indexed components and 90.9% (n=10) of the non-indexed components allowed bacterial leakage, with significant difference between groups (p=0.0237). In conclusion, both tapered components failed to provide adequate sealing to bacterial leakage, although the indexed type components showed a superior seal compared with non-indexed components.

Resumo Conexões de implantes cônicos cresceram em popularidade por serem mais resistentes à fadiga e por promover uma melhor vedação contra infiltração bacteriana do que as conexões convencionais. O objetivo deste estudo foi avaliar o selamento bacteriano na interface implante-pilar utilizando dois modelos de implantes cone Morse, por meio de análise microbiológica in vitro. Onze pilares não indexados e 11 pilares indexados foram selecionados e conectados aos seus respectivos implantes com um torque de 20 N, de acordo com a recomendação do fabricante. A análise microbiológica foi realizada utilizando colônias de Escherichia coli retirados diretamente a partir de uma placa de cultura para o componente protético. Para os grupos de controle, foi utilizado um pilar-implante não contaminado de cada grupo (controle negativo) e um implante contaminado sem pilar (controle positivo). Os espécimes foram imersos em caldo BHI e mantidos numa incubadora a 37 °C durante 14 dias, para monitorar o desenvolvimento de contaminação bacteriana. Os resultados revelaram que 36,4% (n=4) dos componentes indexados e 90,9% (n=10) dos componentes não indexados obtiveram infiltração bacteriana, com diferença significativa entre os grupos (p=0,0237). Como conclusão, os dois componentes cônicos não conseguiram proporcionar uma vedação adequada contra infiltração bacteriana, embora os componentes do tipo indexados mostrassem uma vedação superior, quando comparados com componentes não indexados.

Evaluation of the Biological Behavior of Mucograft® in Human Gingival Fibroblasts: An In Vitro Study

Mucograft(r) is a resorbing porcine matrix composed of type I and type III collagen, used for soft tissue augmentation in guided tissue bony regeneration procedures. This in vitro study aimed to evaluate the biological behavior of Mucograft(r) in human gingival fibroblasts, as well as the ability of the matrix to induce production of extracellular matrix. Six resorbing Mucograft(r) matrices (MCG) were cut into 3 x 2 mm rectangles and 5 x 5 mm squares and were placed in 96- and 24-well plates, respectively. The control group (CTRL) consisted of cells plated on polystyrene without the MCG. After one, two, three and seven days, cell proliferation and viability were assessed using the Trypan exclusion method and MTT test, respectively. Type III collagen (COL 3A1) and vimentin (VIM) expression were also evaluated at 10 and 14 days, using Western blotting. Statistical analysis, using ANOVA with post hoc Bonferroni test, revealed that human gingival fibroblasts from MCG showed similar results (p>0.05) for proliferation and viability as the cells cultured on CTRL. After 14 days, a significant decrease in COL 3A1 expression (p<0.05) was observed when cultured with the MCG. VIM expression showed no significant difference at any time period (p>0.05). Although no increase in extracellular matrix secretion was observed in this in vitro study, Mucograft(r) presented cellular compatibility, being an option for a scaffold whenever it is required.

Resumo A Mucograft(r) é uma matriz reabsorvível, de origem suína, composta de colágenos do tipo I e III, utilizada para aumento de tecido mole em regeneração óssea guiada. Este estudo in vitro teve como objetivo avaliar o comportamento biológico da Mucograft(r), em fibroblastos gengivais humanos, bem como a indução da síntese de matriz extracelular. Seis matrizes reabsorvíveis de Mucograft(r) (MCG) foram cortadas em retângulos e quadrados medindo 3 x 2 mm e 5 x 5 mm e alocadas em placas de 96 e 24 poços, respectivamente. O grupo controle (CTRL) consistiu no plaqueamento celular em poliestireno, sem MCG. Após um, dois, três e sete dias, a proliferação e a viabilidade celular foram avaliadas utilizando o corante vital azul de Trypan e o teste MTT, respectivamente. Além disso, a expressão de colágeno tipo III (COL 3A1) e vimentina (VIM) foi avaliada após 10 e 14 dias, por meio de Western-blotting. Após análise estatística (Anova e pós teste de Bonferroni), pode-se observar que os fibroblastos gengivais humanos, cultivados sobre MCG, apresentaram proliferação e viabilidade semelhantes em comparação às células que foram cultivadas apenas no poliestireno (CTRL). Após 14 dias, notou-se uma diminuição significativa da expressão de COL 3A1 (p<0,05) quando as células foram cultivadas sobre a MCG. A expressão da VIM não mostrou diferença significativa em nenhum dos períodos estudados (p>0,05). No presente estudo in vitro pode-se concluir que apesar de não ter sido observado aumento da síntese de matriz extracelular, a Mucograft(r) apresentou compatibilidade celular, sendo uma opção de biomaterial em casos que o arcabouço é necessário.

Effects of a Mikania laevigata extract on bone resorption and RANKL expression during experimental periodontitis in rats

OBJECTIVES: The Mikania laevigata extract (MLE) (popularly known in Brazil as "guaco") possesses anti-inflammatory properties. In the present study we tested the effects of MLE in a periodontitis experimental model in rats. We also investigated possible mechanisms underlying such effects. MATERIAL AND METHODS: Periodontal disease was induced by a ligature placed around the mandibular first molars of each animal. Male Wistar rats were divided into 4 groups: non-ligated animals treated with vehicle; non-ligated animals treated with MLE (10 mg/kg, daily); ligature-induced animals treated with vehicle and ligature-induced animals treated with MLE (10 mg/kg, daily). Thirty days after the induction of periodontal disease, the animals were euthanized and mandibles and gingival tissues removed for further analysis. RESULTS: Morphometric analysis of alveolar bone loss demonstrated that MLE-treated animals presented a decreased alveolar bone loss and a lower expression of the activator of nuclear factor-κB ligand (RANKL) measured by immunohistochemistry. Moreover, gingival tissues from the MLE-treated group showed decreased neutrophil migration myeloperoxidase (MPO) assay. CONCLUSIONS: These results indicate that MLE may be useful to control bone resorption during progression of experimental periodontitis in rats.

15d-PGJ2 modulates acute immune responses to Trypanosoma cruzi infection

The acute phase of Trypanosoma cruzi infection is associated with a strong inflammatory reaction in the heart characterised by a massive infiltration of immune cells that is dependent on the T. cruzi strain and the host response. 15d-PGJ2 belongs to a new class of anti-inflammatory compounds with possible clinical applications. We evaluated the effects of 15d-PGJ2 administered during the acute phase of T. cruzi infection in mice. Mice were infected with the Colombian strain of T. cruzi and subsequently treated with 15d-PGJ2 repeatedly for seven days. The inflammatory infiltrate was examined by histologic analysis. Slides were immunohistochemically stained to count the number and the relative size of parasite nests. Infection-induced changes in serum cytokine levels were measured by ELISA. The results demonstrated that treatment with 15d-PGJ2 reduced the inflammatory infiltrate in the skeletal muscle at the site of infection and decreased the number of lymphocytes and neutrophils in the blood. In addition, we found that 15d-PGJ2 led to a decrease in the relative volume density of amastigote nests in cardiac muscle. T. cruzi-infected animals treated with 15d-PGJ2 displayed a statistically significant increase in IL-10 levels with no change in IFN-ã levels. Taken together, we demonstrate that treatment with 15d-PGJ2 in the acute phase of Chagas disease led to a controlled immune response with decreased numbers of amastigote nests, as measured by the volume density.

Delay in maturation of the submandibular gland in Chagas disease correlates with lower DNA synthesis

It has been demonstrated that the acute phase of Trypanosoma cruzi infection promotes several changes in the oral glands. The present study examined whether T. cruzi modulates the expression of host cell apoptotic or mitotic pathway genes. Rats were infected with T. cruzi then sacrificed after 18, 32, 64 or 97 days, after which the submandibular glands were analyzed by immunohistochemistry. Immunohistochemical analyses using an anti-bromodeoxyuridine antibody showed that, during acute T. cruzi infection, DNA synthesizing cells in rat submandibular glands were lower than in non-infected animals (p < 0.05). However, after 64 days of infection (chronic phase), the number of immunolabeled cells are similar in both groups. However, immunohistochemical analysis of Fas and Bcl-2 expression did not find any difference between infected and non-infected animals in both the acute and chronic stages. These findings suggest that the delay in ductal maturation observed at the acute phase of Chagas disease is correlated with lower expression of DNA synthesis genes, but not apoptotic genes.

Ação dos linfócitos T regulatórios em transplantes: [revisão] / T regulatory lymphocytes action in transplants: [review]

O sistema imunológico humano possui um significado comum de proteção contra antígenos estranhos com potencialidade patogênica ou não, ativando uma ação coletiva e coordenada entre células e moléculas. Embora sendo um sistema benéfico, deve ser controlado para evitar que auto-antígenos sejam alvejados. Recentemente, por meio de novas técnicas, tem-se observado uma corrente de estudo para as células T CD4+CD25+ Foxp3 como células reguladoras que controlam ativamente a função de outras células imunes impedindo sua atividade e, conseqüentemente, o desenvolvimento de doenças auto-imunes, rejeição de enxerto e combate a células tumorais. Em pacientes submetidos ao transplante alogênico, os mecanismos que levam a hiporresponsividade, assim como os mecanismos que permitem uma maior sobrevida do enxerto ainda são pouco conhecidos. Este artigo aborda uma revisão da literatura sobre as células T regulatórias, vislumbrando uma nova possibilidade de terapia imunomoduladora para pacientes transplantados.

The human immune system mounts specific responses against a vast array of foreign antigens, pathogenic or otherwise, activating a coordinated action between cells and molecules. Although this is beneficial, it must be carefully controlled to ensure that normal self antigens are not targeted. Recently, with the development of new techniques, it has been observed that T CD4+CD25+Foxp3 act as regulatory cells which actively control the properties of other immune cells by suppressing their functional activity to prevent autoimmunity and transplant rejection as well as to trigger the immune system against tumor cells. In patients submitted to allogeneic transplantation, specific unresponsive mechanisms and mechanisms that induce and maintain graft tolerance are little understood. This article reviews what is currently known about these so-called regulatory T cells and discusses the potential use of these cells in transplantation immunolog.

An in vitro microbial model for producing caries-like lesions on enamel

This study aimed to develop a low cost in vitro viable microbiological model to produce biofilms to be used in dental researches. Single and multi-species biofilms of S. mutans, S. sobrinus, S. mitis, S. salivarius, S. cricetus and S. sanguinis were grown on bovine enamel slabs during 10 days, in a sterile brain-heart infusion broth, containing 5% sucrose and incubated at 37ºC in an atmosphere of 10% CO2. The slabs were transferred to a fresh medium at every 6, 12 or 24 hours. After the experimental period, enamel volume percent mineral was determined by cross-sectional microhardness. Caries-like lesions were found in all bacterial groups when compared with the control group. No statistical significant differences were found between S. mutans and S. sobrinus with respect of their cariogenicity or among the periods of medium change. However, it was found a statistical significant difference among the cariogenicity of S. salivarius and S. sanguinis (ANOVA followed by Tukey test). This model has successfully developed caries-like lesion on enamel and the medium can be changed at every 24 hours utilizing either S. mutans or S. sobrinus.