Overview of miRNAs for the non-invasive diagnosis of endometriosis: evidence, challenges and strategies. A systematic review / Visão geral de miRNAs como diagnóstico não invasivo de endometriose: evidências, desafios e estratégias. Uma revisão sistemática

ABSTRACT Objective The aim of the study was to assess the evidence on miRNAs as biomarkers for the diagnosis of endometriosis, as well as to provide insights into the challenges and strategies associated with the use of these molecules as accessible tools in clinical practice. Methods Systematic review conducted on PubMed®, Latin American and Caribbean Health Sciences Literature (LILACS), MEDLINE® and Web of Science databases using the search terms endometriosis (all fields) AND miRNA (all fields), evaluating all publication up to May 2019. Results Most miRNAs found to be dysregulated in this study were harvested from tissue samples, which precludes their use as a non-invasive diagnostic test. However, differential expression of 62 miRNAs was reported in samples that may be used for non-invasive diagnosis of endometriosis, such as blood, serum and plasma. Conclusion Despite the identification of several candidates, studies are investigatory in nature and have been conducted with small number of samples. Also, no particular miRNA has been validated for diagnostic purposes so far. Studies based primarily on biological samples and applicable to translational research are warranted. Large databases comprising information on sample type and the use of saliva and vaginal fluid for miRNAs identification may prove essential to overcome current barriers to diagnosis of endometriosis.

RESUMO Objetivo O objetivo do estudo foi analisar as evidências sobre miRNAs como biomarcadores para o diagnóstico de endometriose, bem como levantar informações sobre os desafios e as estratégias necessárias para tornar essas moléculas ferramentas acessíveis para uso na prática clínica. Métodos Revisão sistemática conduzida nos bancos de dados PubMed®, Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), MEDLINE® e Web of Science utilizando os termos de pesquisa "endometriosis" (todos os campos) AND "miRNA" (todos os campos), avaliando todas as publicações até maio de 2019. Resultados A maioria dos miRNAs desregulados foram analisados a partir de amostras de tecido, o que inviabiliza seu uso como teste diagnóstico não invasivo. Todavia, 62 miRNAs foram identificados como diferencialmente expressos em amostras que poderiam ser usadas para o diagnóstico pouco invasivo de endometriose, como sangue, soro e plasma. Conclusão Apesar de todos esses candidatos, os trabalhos são exploratórios, realizados com números pequenos de amostras, sem miRNAs específicos validados para fins diagnósticos. Estudos envolvendo principalmente amostras biológicas, visando à pesquisa translacional, deveriam ser mais explorados. O desenvolvimento de grandes bancos de dados sobre amostras, bem como o uso de saliva e fluido vaginal para identificação de miRNAs, poderia servir como recursos essenciais para as barreiras atuais no diagnóstico da endometriose.

Long-term evaluation of the antimicrobial susceptibility and microbial profile of subgingival biofilms in individuals with aggressive periodontitis

This study evaluates the antimicrobial susceptibility and composition of subgingival biofilms in generalized aggressive periodontitis (GAP) patients treated using mechanical/antimicrobial therapies, including chlorhexidine (CHX), amoxicillin (AMX) and metronidazole (MET). GAP patients allocated to the placebo (C, n = 15) or test group (T, n = 16) received full-mouth disinfection with CHX, scaling and root planning, and systemic AMX (500 mg)/MET (250 mg) or placebos. Subgingival plaque samples were obtained at baseline, 3, 6, 9 and 12 months post-therapy from 3–4 periodontal pockets, and the samples were pooled and cultivated under anaerobic conditions. The minimum inhibitory concentrations (MICs) of AMX, MET and CHX were assessed using the microdilution method. Bacterial species present in the cultivated biofilm were identified by checkerboard DNA-DNA hybridization. At baseline, no differences in the MICs between groups were observed for the 3 antimicrobials. In the T group, significant increases in the MICs of CHX (p < 0.05) and AMX (p < 0.01) were detected during the first 3 months; however, the MIC of MET decreased at 12 months (p < 0.05). For several species, the MICs significantly changed over time in both groups, i.e., Streptococci MICs tended to increase, while for several periodontal pathogens, the MICs diminished. A transitory increase in the MIC of the subgingival biofilm to AMX and CHX was observed in GAP patients treated using enhanced mechanical therapy with topical CHX and systemic AMX/MET. Both protocols presented limited effects on the cultivable subgingival microbiota.

Characteristics of secondary caries lesion around composite resin restorations / Características da cárie secundária adjacente a restaurações de resinacomposta

Objetivo: O objetivo deste estudo foi analisar as características de cárie secundária e sua relação com a presença de restaurações com defeitos marginais como gap, degrau negativa (NL, a falta de material restaurador) e degrau positiva (PL, saliência). Materiais e Métodos: Oitenta superfícies proximais de restaurações de resina classe II foram fotografados e analisados utilizando um microscópio estereoscópico (X40). Foi registrada a presença de cáries secundárias externas, gaps, NL, PL e após a remoção de restaurações, a presença de lesão interna. Todos os exames foram realizados em 10 pontos bem definidos. Resultados: A prevalência de cárie secundária e gap foram associados com margem gengival. As restaurações com defeitos marginais (NL e PL) não foram associados a qualquer local específico (vestibular, lingual ou gengival). Observou-se uma relação entre esses defeitos com a prevalência de cárie secundária. A presença da lesão interna foi associada com lesão externa. Embora, em 141 pontos foi observada lesão interna, sem a presença de lesão externa. A maioria destas lesões mostraram envolvimento de esmalte interno. Apenas 8 por cento tinham desmineralização restrito a dentina, que poderia sugerir cárie residuais.

Objective: The aim of this study was to analyze the features of secondary caries and their relationship with the presence of restorations marginal defects as gap, negative ledge (NL, lack of restorative material) and positive ledge (PL, overhang). Materials and methods: Eighty proximal surfaces of Class II resin restorations were photographed and analyzed using a stereomicroscope (X40). It was recorded the presence of external secondary caries, gaps, NL, PL and after the restorations removal, the presence of internal lesion. All the exams were done in 10 welldefined points. Results: The prevalence of secondary caries and gap were associated with gingival margin. The restorations marginal defects (NL and PL) were not associated with any specific location (buccal, lingual or gingival). It was observed a relationship between these defects with secondary caries prevalence (gap: p= 0.004; NL and PL: p= 0.017). This association could be explained by the biofilm accumulation usually observed in the interface tooth surface/ restoration marginal defects. The presence of internal lesion was associated with external lesion. Although, in 141 points was observed internal lesion without the presence of external lesion. The majority of these lesions showed internal enamel involvement. Only 8 percent had demineralization restricted to dentine, which could suggest residual caries. Conclusion: The results of this study showed that secondary caries lesion frequently began externally on the interface tooth/restoration, where diagnose, control and arrestment of the lesions is possible.

Histological features of initial secondary caries adjacent to composite resin restorations / Características Histológicas da Lesão Inicial da Cárie Secundária Adjacente a Restaurações de Resina Composta

O objetivo deste estudo foi analisar morfologicamente o início da lesão de cárie secundária. Trinta e três superfícies proximais adjacentes a restaurações de resina classe II foram examinadas ao estereomicroscópio (40x) para a presença de carie e gap. Sessenta sítios (39 sadios, 7 com lesões, 14 adjacentes a lesões) foram preparados para exame utilizando microscopia de luz polarizada. As lesões foram observadas em 52,7% dos corte úmidos e 85,0% dos cortes secos. Os cortes secos demonstraram lesões maiores. Analises posteriores foram realizadas nos cortes secos. Cem por cento das secções observadas no estereomicroscópio, 87,2% das lesões hígidas, e 71,4% dos sítios adjacentes a lesões demonstraram áreas desmineralizadas. A análise morfológica indicou que a lesão cariosa inicia na superfície de esmalte externa na interface entre dente e restauração. Dois padrões distintos de desenvolvimento de carie foram observados, relacionados à direção dos prismas de esmalte: (1) a direção dos prismas no esmalte adjacente foi paralela à parede da cavidade (a parte mais profunda da penetração da cárie foi localizada na parede da cavidade), e (2) a direção dos prismas corria na direção apical (a parte mais profunda da penetração da cárie foi obliqua à parede da cavidade). O padrão 1 pode ser confundido por uma parede da lesão. A prevalência de lesões adjacentes às restaurações foi similar em sítios com (85,7%) e sem (84,0%) gaps. Os achados suportam o pressuposto de que o inicio da lesão de cárie secundária in vivo não está relacionado à presença de um gap e segue a direção dos prismas de esmalte.

The aim was to analyze morphologically the initiation of secondary caries lesion. 33 proximal surfaces adjacent to Class II resin restorations were examined by stereomicroscope (x40) for the presence of caries and gap. 60 sites (39 sound, 7 with lesions, 14 adjacent to lesions) were prepared for examination using polarized light microscopy. Lesions were observed in 52.7% of the wet sections and 85.0% of the dried sections. Dried ground sections displayed larger lesions. Further analyses were performed on dried sections. 100% of ground sections from the stereomicroscope carious sites, 87.2% from sound sites, and 71.4% from sites adjacent to lesions displayed demineralized areas. Morphological analysis indicated that caries lesion initiates at the external enamel surface in the interface between tooth and restoration. Two distinct patterns of caries development were observed, related to the direction of enamel rods: (1) the direction of the rods in the adjacent enamel was parallel to the cavity wall (the deepest part of caries penetration was located at the cavity wall), and (2) the direction of the rods ran in the apical direction (the deepest part of caries penetration was oblique to the cavity wall). The pattern 1 could be mistaken for a wall lesion. The prevalence of lesions adjacent to restorations was similar in the sites with (85.7%) and without (84.0%) gaps. The findings support the assumption that secondary caries initiation in vivo is not related to the presence of a gap and follows the direction of the enamel rods.