Epithelial responses and Candida albicans pathogenicity are enhanced in the presence of oral streptococci

Abstract Experimental models that consider host-pathogen interactions are relevant for improving knowledge about oral candidiasis. The aim of this study was to assess the epithelial immune responses, Candida penetration of cell monolayers, and virulence during mixed species culture infections. Single species cultures of Candida albicans and mixed cultures (C. albicans, Streptococcus mutans, and Streptococcus sanguinis) were used to infect monolayers of HaCaT and FaDu ATCC HTB-43 cells for 12 h. After infection, IL-18 and IL-34 gene expression was measured to assess epithelial cell immune responses, and lactate dehydrogenase (LDH) activity was measured as an indicator of cell damage. Microscopy determined C. albicans morphology and penetration of fungal cells through the keratinocyte monolayer. Monolayers devoid of infection served as controls. Data were analyzed by an ANOVA one-way test followed by Tukey's post-hoc test (α = 0.05). The results found that IL-18 and IL-34 gene expression and LDH activity were significantly (p < 0.05) upregulated for both cell lines exposed to mixed species cultures compared with C. albicans alone. Candida albicans yeast and hyphae were evident in C. albicans only infections. In contrast, monolayers infected by C. albicans, S. mutans, and S. sanguinis exhibited higher microbial invasion with several hyphal aggregates detected. The presence of streptococci in C. albicans infection enhances the virulence and pathogenicity of the fungus with associated increased immune responses and tissue damage. Extrapolation of these findings to oral infection would indicate the added potential benefit of managing bacterial components of biofilms during treatment.

Resumo O objetivo deste estudo foi avaliar a resposta epithelial imune, a colonização da Candida albicans em monocamadas celulares e sua virulência em resposta a infecções de culturas de biofilme multiespécie. Culturas de biofilme monoespécie de C. albicans e culturas mistas (C. albicans, Streptococcus mutans e Streptococcus sanguinis) foram utilizadas para infectar monocamadas de células HaCaT e FaDu por 12 h. Após a infecção, a expressão dos genes IL-18 e IL-34 foi medida para avaliar as respostas imunes das células epiteliais. A atividade da lactato desidrogenase (LDH) foi medida como um indicador de dano celular. A microscopia determinou a morfologia de C. albicans e a penetração das células fúngicas através da monocamada de queratinócitos. Monocamadas em que não houve infecção serviram como controles. Os dados foram analisados por um teste ANOVA one-way seguido pelo teste post-hoc de Tukey (α = 0,05). Os resultados demonstraram que a expressão gênica de IL-18 e IL-34 e a atividade de LDH foram (p < 0,05) reguladas positivamente para ambas as linhagens de células expostas a culturas de espécies mistas em comparação com C. albicans isoladamente. Leveduras de C.albicans e hifas foram evidentes em infecções apenas por C. albicans. Entretanto, monocamadas infectadas por C. albicans, S. mutans e S. sanguinis exibiram maior invasão microbiana com vários agregados de hifas detectados. Dessa maneira, a presença de estreptococos na infecção por C. albicans aumentou a virulência e a patogenicidade do fungo com respostas imunes aumentadas associadas a danos nos tecidos. A extrapolação desses achados para a infecção oral indicaria o potencial benéfico do controle dos componentes bacterianos em biofilmes durante a terapia da candidíase

Biological properties of Schinus terebinthifolia Raddi essential oil

Abstract Schinus terebinthifolia Raddi green fruits essential oil (EO) was evaluated regarding its phytochemical profile, antimicrobial and cytotoxic activities, and toxicity. Gas chromatography with mass spectrometry was applied to identify its constituents, thereafter the minimum inhibitory concentration, minimum bactericidal and fungicidal concentrations, and its antibiofilm activity were evaluated. The EO cytotoxicity was assessed in tumor and non-tumor human cells, and in vivo toxicity was evaluated in a Galleria mellonella model. The major constituents of S. terebinthifolia EO were alpha-phellandrene and beta-phellandrene. The EO had a weak activity against all strains of Candida albicans (MIC 1000µg/mL) and had no activity against non-albicans strains, bacteria, and C. albicans biofilm. Cytostatic activity against all tumor cell lines was shown. Additionally, cell viability remained at EO concentrations up to 62.5 µg/mL. At 16 mg/mL, 50% hemolysis was observed, and it had low toxicity in vivo. Overall, the S. terebinthifolia EO was characterized by low antimicrobial and antibiofilm activities, with no evidence of toxicity to human tumor and non-tumor cells

Adesão de biofilmes monoespécie de Streptococcus mutans e Candida albicans em diferentes superfícies de resinas compostas convencionais e bulk fill / Adhesion of monospecies biofilms from Streptococcus mutans and Candida albicans to different surfaces of conventional composite resins and bulk fill

Introdução: As resinas compostas são alternativas restauradoras, porém sua superfície pode favorecer o acúmulo de biofilme. Objetivo: Analisar in vitro a adesão de biofilmes de Streptococcus mutans (UA159) e Candida albicans (ATCC 90028) em superfícies de resinas compostas convencionais e bulk fill. Material e método: Foram utilizadas quatro marcas de resinas compostas e bulk fill: Aura Bulk Fill - SDI®; Premisa - Kerr®; Opallis- FGM®, e Filtek bulk fill flow - 3M®. Utilizou-se saliva artificial para formação da película salivar, por 60 min a 37 °C. O inóculo foi padronizado em 1×108 UFC/mL para S. mutans e 1×106 UFC/mL para C. albicans. Os espécimes (n=8/grupo) foram acondicionados em placas de 24 poços, com BHI suplementado com sacarose para as bactérias, e RPMI 1640, para os fungos. A formação do biofilme foi avaliada considerando as unidades formadoras de colônia (UFC/mL).Os dados foram analisados por ANOVA e Tukey (p<0,05). Resultado: Para os biofilmes de S. mutans, não houve diferença significativa na contagem de UFC/mL entre os diferentes tipos de resina (p=0,119). Na contagem de UFC/mL para biofilme de Candida, as médias variaram entre 7,78 e 8,34. Houve diferença significativa entre as marcas, especialmente entre as resinas convencionais e bulk fill. Conclusão: O presente estudo demonstra que não há diferença na adesão para biofilmes de S. mutans. Porém, há diferença na adesão da C. albicans na superfície de diferentes resinas compostas.

Introduction: Composite resins are restorative alternatives, but their surface may favor the accumulation of biofilm. Objective: The aim of this study was to evaluate the in vitro adhesion of Streptococcus mutans (UA159) and Candida albicans (ATCC 90028) biofilms on the surface of conventional and bulk fill composites. Material and method: Four brands of conventional and bulk fill composites were used, Aura Bulk Fill - SDI®, Premisa - Kerr®, Opallis- FGM® and Filtek bulk fill flow - 3M®. Artificial saliva was used to form the salivary film for 60 min at 37 ° C. The inoculum was standardized at 1x108 CFU/mL for S. mutans and 1x106 CFU/mL for C. albicans. The specimens (n = 8/group) were placed in 24-well plates, with BHI supplemented with sucrose for bacteria and RPMI 1640 for fungi. The biofilm formation was evaluated considering the colony forming units (CFU/mL). The data were analyzed by ANOVA and Tukey test (p <0.05). Result: For S. mutans biofilms, there was no significant difference in the CFU/mL count between the different types of composites (p = 0.119). In the CFU / mL count for C.albicans biofilm, the CFU/mL ranged from 7.78 to 8.34. There was a significant difference between brands for Candida, especially between conventional and bulk fill composites. Conclusion: The present study demonstrates that there is no difference in adhesion for S. mutans biofilms. On the other hand, there is a difference in the adhesion of C. albicans to the surface of different composite resins.

Offer and Use of Oral Health in Primary Care Before and After the Beginning of the COVID-19 Pandemic in Brazil

Abstract Objective: To compare the offer and use of oral health services in primary care, before and after the beginning of the COVID-19 pandemic in Brazil. Material and Methods: An observational study with a cross-sectional ecological design, using data from the Health Information System for Primary Care. Data regarding the number of Oral Health Teams (OHT), Oral Health Coverage in Primary Care (OHC), number of First Programmatic Dental Consultations (FPDC), and number of visits due to dental abscess and toothache were collected. Data regarding the 26 Brazilian states and Federal District were collected, as consolidated of the first quarter (January to April) of 2019 and of 2020. The median of the difference (MD) and the percentage of variation (%V) were obtained for each variable and were compared by Wilcoxon test (α<0.05). Results: An increase in the number of OHT was observed in 25 states (MD=45; %V=6.13; p<0.001), whilst the OHC increased in 17 states (MD=1.01; %V=1.62; p=0.035) between the 2019 and 2020. We also verified a significant reduction in the number of FPDC (MD=- 42.806; %V=-38.70; p<0.001), as well as in the number of visits due to dental abscess (MD=-1.032; %V=-29.04; p=0.002) and due to toothache (MD=-14.445; %V=-32.68; p<0.001). Conclusion: Although an expansion of OHT and OHC between 2019 and 2020 was verified, the offer and use of oral health services in primary care has decreased due to the COVID-19 pandemic.

Dental caries experience and associated factors in 12-year-old-children: a population based-study

Abstract Relevant public policies in oral health have been implemented in Brazil since 2004. Changes in the epidemiological status of dental caries are expected, mainly in the child population. This study aimed to assess the dental caries experience and associated factors among 12-year-old children in the state of São Paulo, Brazil. A cross-sectional population-based study was conducted with data obtained from 415 cities, including a total of 26,325 schoolchildren who were included by the double-stage cluster technique, by lot and by systematic sampling. The statistical model included data from the São Paulo Oral Health Survey (SBSP 2015), the Human Development Index (HDI) and the Social Vulnerability Index (SVI). A theoretical-conceptual model categorized the variables into three blocks, namely, contextual (HDI, SVI, region of residence and fluoridation of water), individual (sex and ethnicity) and periodontal conditions (gingival bleeding, dental calculus and the presence of periodontal pockets), for association with the experience of caries (DMFT). Statistically significant associations were verified by hierarchical multivariate logistic (L) and Poisson (P) regression analyses (p < 0.05). The results showed that 57.7% of 12-year-old children had caries experience. Factors that determined a greater prevalence of dental caries in both models were nonwhite ethnicity (ORL = 1.113, ORP = 1.154) and the presence of gingival bleeding (ORL = 1.204, ORP = 1.255). Male children (ORL = 0.920 ORP = 0.859) and higher HDI (ORL = 0.022), ORP = 0.040) were associated with a lower prevalence of dental caries experience. Water fluoridation was associated with a lower DMFT index (ORP = 0.766). Dental caries experience is still associated with social inequalities at different levels. Policymakers should direct interventions towards reducing inequalities and the prevalence of dental caries among 12-year-old children.

Adenocarcinoma polimorfo de baixo grau de grandes proporções em paciente idoso: relato de caso não usual / Large polymorphous low-grade adenocarcinoma in elderly patient: unusual case report

O adenocarcinoma polimorfo de baixo grau (APBG) é uma neoplasia maligna que ocorre predominantemente em glândulas salivares menores e exibe baixa tendência para o comportamento agressivo e a disseminação metastática. O presente trabalho objetiva relatar um caso de APBG em paciente idoso que se configurou como não usual, especialmente pela grande dimensão apresentada pela lesão. Adicionalmente, serão abordados os principais critérios para um correto diagnóstico diferencial com outras neoplasias de glândulas salivares. Paciente, 92 anos, sexo masculino, tabagista, apresentava um tumor assintomático na região esquerda do palato, recoberto por mucosa de coloração normal com áreas telangiectásicas, medindo aproximadamente 7,4 cm. Sob hipótese diagnóstica de carcinoma mucoepidermoide foi realizada uma biópsia do tipo incisional. Após análise histopatológica, os achados microscópicos foram compatíveis com APBG e o paciente foi encaminhado para o cirurgião de cabeça e pescoço. O APBG é uma neoplasia maligna de glândulas salivares que exibe baixa tendência para o comportamento agressivo e se apresenta, geralmente, como um nódulo indolor. No entanto, como o APBG pode alcançar grandes dimensões, os autores ressaltam a importância de um criterioso diagnóstico deste tumor, a fim de diferenciá-lo de outras neoplasias de comportamento biológico e prognóstico diferentes

Polymorphous low-grade adenocarcinoma (PLGA) is a malignant tumor which occurs predominantly in minor salivary glands and exhibits low tendency for aggressive behavior and metastatic spread. The aim of this paper is to report an unusual case of PLGA in an elderly patient, specially because of the large size of the lesion. Furthermore, the main criteria for differential diagnosis between PLGA and other salivary gland neoplasms will be discussed. A 92-year-old male patient, who was a smoker, presented with an asymptomatic tumor on the left portion of the palate, covered by normal mucosa with telangiectatic areas, measuring about 7.4 cm. The diagnostic hypothesis was mucoepidermoid carcinoma and incisional biopsy was done. Histopathologic features were compatible with PLGA and the patient was referred to the head and neck surgeon. PLGA is a malignant salivary gland neoplasm that exhibits low tendency for aggressive behavior and usually presents as a non-tender nodule. Nevertheless, since PLGA can reach a large size, the authors emphasize the importance of an accurate diagnosis of this tumor in order to differentiate it from other neoplasms with different biological behavior and prognosis