RESUMO
The accumulated dental biofilm can be a source of oral bacteria that are aspirated into the lower respiratory tract causing ventilator-associated pneumonia in hospitalized patients. The aim of this study was to evaluate the synergistic antibiofilm action of the produced and phytochemically characterized extracts of Cinnamomum verum and Brazilian green propolis (BGP) hydroethanolic extracts against multidrug-resistant clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition to their biocompatibility on human keratinocyte cell lines (HaCaT). For this, High-performance liquid chromatography analysis of the plant extracts was performed; then the minimum inhibitory and minimum bactericidal concentrations of the extracts were determined; and antibiofilm activity was evaluated with MTT assay to prevent biofilm formation and to reduce the mature biofilms. The cytotoxicity of the extracts was verified using the MTT colorimetric test, evaluating the cellular enzymatic activity. The data were analyzed with one-way ANOVA and Tukey's tests as well as Kruskal-Wallis and Dunn's tests, considering a significance level of 5%. It was possible to identify the cinnamic aldehyde in C. verum and p-coumaric, caffeic, and caffeoylquinic acids as well as flavonoids such as kaempferol and kaempferide and Artepillin-C in BGP. The combined extracts were effective in preventing biofilm formation and reducing the mature biofilms of A. baumannii and P. aeruginosa. Moreover, both extracts were biocompatible in different concentrations. Therefore, C. verum and BGP hydroethanolic extracts have bactericidal and antibiofilm action against multidrug resistant strains of A. baumannii and P. aeruginosa. In addition, the combined extracts were capable of expressively inhibiting the formation of A. baumannii and P. aeruginosa biofilms (prophylactic effect) acting similarly to 0.12% chlorhexidine gluconate.
Assuntos
Acinetobacter baumannii , Própole , Humanos , Pseudomonas aeruginosa , Própole/farmacologia , Cinnamomum zeylanicum , Brasil , Farmacorresistência Bacteriana Múltipla , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Biofilmes , QueratinócitosRESUMO
Candida spp. cause fungal infection that affects patients' oral health. This study aimed to evaluate the isolated and synergistic antifungal effect of Rosa centifolia L., Curcuma longa L., Rosmarinus officinalis L., and Punica granatum L. glycolic extracts against Candida albicans, Candida dubliniensis, Candida tropicalis, and Candida krusei planktonic and biofilm forms. The plant extracts were chemically characterized and the main compounds were quantified by high-performance liquid chromatography (HPLC-DAD) analysis. The minimum inhibitory and minimum fungicidal concentrations of the extracts were determined, and antibiofilm activity was evaluated by MTT assay. Data were analyzed by one-way ANOVA and Tukey's tests, and by Kruskal-Wallis and Dunn's tests, considering a significance level of 5%. The main compounds identified in each of the extracts were: p-coumaric acid (2153.22 µg/100 mL) in the rosemary extract, gallotannins (4318.31 µg/100 mL) in the pomegranate extract, quercetin derivatives (3316.50 µg/100 mL) in the extract of white roses, and curcumin (135.09 µg/100 mL) in the turmeric extract. The combination of R. centifolia and C. longa glycolic extracts was effective against C. albicans, C. dubliniensis, and C. tropicalis biofilms over different periods (p < 0.05). The combination of R. officinalis and P. granatum glycolic extracts was effective against C. albicans and C. krusei biofilms after 30 min, and against C. tropicalis after 24 h, with all combinations showing an average reduction of 50% in cell viability (p < 0.05). In conclusion, the combined plant extracts have antifungal and antibiofilm action against Candida spp. in different concentrations and times of action.
Assuntos
Antifúngicos , Glicóis , Humanos , Antifúngicos/química , Candida , Candida albicans , Candida tropicalis , Extratos Vegetais/química , Testes de Sensibilidade Microbiana , BiofilmesRESUMO
OBJECTIVE: To evaluate the chemical composition and effects of Artemisia vulgaris (AV) hydroalcoholic extract (HEAV) on breast cancer cells (MCF-7 and SKBR-3), chronic myeloid leukemia (K562) and NIH/3T3 fibroblasts. METHODS: Phytochemical analysis of HEAV was done by high-performance liquid chromatography-mass (HPLC) spectrometry. Viability and cell death studies were performed using trypan blue and Annexin/FITC-7AAD, respectively. Ferrostatin-1 (Fer-1) and necrostatin-1 (Nec-1) were used to assess the mode of HEAV-induced cell death and acetoxymethylester (BAPTA-AM) was used to verify the involvement of cytosolic calcium in this event. Cytosolic calcium measurements were made using Fura-2-AM. RESULTS: HEAV decreased the viability of MCF-7, SKBR-3 and K562 cells (P<0.05). The viability of HEAV-treated K562 cells was reduced compared to HEAV-exposed fibroblasts (P<0.05). Treatment of K562 cells with HEAV induced cell death primarily by late apoptosis and necrosis in assays using annexin V-FITC/7-AAD (P<0.05). The use of Nec-1 and Fer-1 increased the viability of K562 cells treated with HEAV relative to cells exposed to HEAV alone (P<0.01). HEAV-induced Ca2+ release mainly from lysosomes in K562 cells (P<0.01). Furthermore, BAPTA-AM, an intracellular Ca2+ chelator, decreased the number of non-viable cells treated with HEAV (P<0.05). CONCLUSIONS: HEAV is cytotoxic and activates several modalities of cell death, which are partially dependent on lysosomal release of Ca2+. These effects may be related to artemisinin and caffeoylquinic acids, the main compounds identified in HEAV.
RESUMO
Antimicrobial resistance is a worldwide health problem and patients in intensive care are more vulnerable, requiring strict control measures and early identification. Currently, clinical culture materials are used to identify the bacterial agent, but saliva culture is not validated, which has great clinical relevance because it participates in several pathophysiological processes. The aim of this study was to validate saliva culture in an intensive care unit environment, determining its diagnostic value for infection. For this purpose, the results of the 39-month surveillance cultures, from the database of a private hospital were evaluated. A total of 323 cultures were paired between saliva, tracheal secretions, blood and urine from patients who were hospitalized for more than 5 days. The search for correlations between the results was performed using the Spearman correlation test. Severity and evolution data were also correlated. It was possible to correlate the presence of Klebsiella spp. between blood culture and saliva culture in 25% of the results (r = 0.01) and the correlation between saliva and tracheal secretion was 33% (r = 0.33447) with p < 0.0001. In conclusion, saliva can be an excellent discriminator of systemic infections, and can be considered a useful culture in clinical practice.
Assuntos
Antibacterianos , Saliva , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Klebsiella , Unidades de Terapia IntensivaRESUMO
This study was performed to evaluate the biocompatibility and antifungal effect of Rosmarinus officinalis against Candida albicans in Galleria mellonella model. Five different concentrations of R. officinalis glycolic extract (50; 25; 12.5 e 6.25 mg/mL) were used to evaluate its biocompatibility in G. mellonella model, in which the nystatin suspension (100; 50; 25; 12.5 e 6.25%) was used as a control group. The antifungal action of R. officinalis glycolic extract was evaluated on C. albicans for 72, 48 and 12 h at two different phases: (1) using the extract as therapeutic agent; and (2) using the extract as prophylactic agent. PBS was used as a negative control group. G. mellonella survival curves were plotted using the Kaplan-Meier method and statistical analysis was performed using the log-rank test (Mantel-Cox) and the significance level was set at (α ≤ 0.05). There was no significant difference among the groups in which all were biocompatible except of a significant death rate of 26.6% with nystatin 100%. In phase 1, it was found that after 7 days, there was no statistically significant difference among the prophylactic treatment groups. In phase 2, the groups of R. officinalis 6.25 mg/mL for 72 h and R. officinalis of 12.5 mg/mL for 24 h promoted the survival rate of the larvae in comparison with the control group with a significant difference (p = 0.017) and (p = 0.032) respectively. Therefore, R. officinalis extract is biocompatible in different concentrations and can be used as a prophylactic agent against fungal infection.
Assuntos
Mariposas , Rosmarinus , Animais , Antifúngicos/farmacologia , Candida albicans , Nistatina/farmacologia , Extratos Vegetais/farmacologiaRESUMO
Um dos principais objetivos do tratamento endodôntico é a limpeza dos canais radiculares e o desenvolvimento de novas soluções irrigadoras é de grande importância. Os fitoterápicos são grandes aliados nesse processo pois apresentam inúmeros efeitos biológicos que favorecem o sucesso da terapia endodôntica. O objetivo deste estudo foi realizar uma revisão integrativa da literatura a fim de fazer um levantamento sobre o estado atual da temática "Endodontia e plantas medicinais", buscando avaliar quais os principais fitoterápicos que apresentam resultados promissores para serem aplicados na endodontia. A busca de artigos foi feita nas bases de dados BVS, Scielo, PubMed e Google Acadêmico, com os descritores em inglês e português: "herbal medicine", "dentisty", "phytotherapy" e "endodontics" combinados pelo operador booleano "AND". Os dez principais fitoterápicos que se destacaram foram: própolis, aroeira-da-praia, gengibre, extrato de alho, malva, camomila, copaíba, romã, unha-de-gato e cravo-da-Índia. E as principais atividades biológicas descritas que favorecem o tratamento endodôntico foram a atividade anti-inflamatória, antibacteriana, antisséptica, cicatrizante, analgésica e antifúngica. Conclui-se que os fitoterápicos são uma boa opção para continuação dos estudos científicos e podem ser considerados potentes candidatos para o desenvolvimento de uma formulação para irrigação ou medicação intracanal com futura aplicação nos tratamentos endodônticos.
One of the main goals of endodontic treatment is root canal cleaning and the development of new irrigation solutions is of great importance. Phytotherapeutics are great allies in this process because they have numerous biological effects that favor the success of endodontic therapy. The aim of this study was to carry out an integrative literature review in order to survey the current status of the topic "Endodontics and medicinal plants", seeking to evaluate which are the main phytotherapeutics that present promising results to be applied in endodontics. The search for articles was conducted in the VHL, Scielo, PubMed and Google Academic databases, with the English and Portuguese descriptors "herbal medicine", "dentistry", "phytotherapy" and "endodontics" combined by the Boolean operator "AND". The top ten herbal medicines that stood out were: propolis, mastic, ginger, garlic extract, mallow, chamomile, copaiba, pomegranate, cat's claw, and clove. And the main biological activities described that favor the endodontic treatment were the anti-inflammatory, antibacterial, antiseptic, healing, analgesic and antifungal activity. It is concluded that phytotherapeutics are a good option for further scientific studies and can be considered potent candidates for the development of a formulation for irrigation or intracanal medication with future application in endodontic treatments.
Uno de los principales objetivos del tratamiento endodóntico es la limpieza del conducto radicular y el desarrollo de nuevas soluciones de irrigación es de gran importancia. Los fitoterápicos son grandes aliados en este proceso ya que poseen numerosos efectos biológicos que favorecen el éxito de la terapia endodóntica. El objetivo de este estudio fue realizar una revisión bibliográfica integradora para relevar el estado actual del tema "Endodoncia y plantas medicinales", buscando evaluar cuáles son los principales fitoterápicos que presentan resultados promisorios para ser aplicados en endodoncia. La búsqueda de artículos se realizó en las bases de datos BVS, Scielo, PubMed y Google Académico, con los descriptores en inglés y portugués "herbal medicine", "dentistry", "phytotherapy" y "endodontics" combinados por el operador booleano "AND". Las diez principales hierbas medicinales que destacaron fueron: propóleo, lentisco, jengibre, extracto de ajo, malva, manzanilla, copaiba, granada, uña de gato y clavo de olor. Y las principales actividades biológicas descritas que favorecen el tratamiento endodóntico fueron la actividad antiinflamatoria, antibacteriana, antiséptica, cicatrizante, analgésica y antifúngica. Se concluye que los fitoterápicos son una buena opción para futuros estudios científicos y pueden ser considerados potentes candidatos para el desarrollo de una formulación para irrigación o medicación intracanal con futura aplicación en tratamientos endodónticos.
RESUMO
A análise facial consiste em uma etapa importante do diagnóstico e avaliação dos resultados obtidos na harmonização orofacial. Para avaliação dos resultados de procedimentos estéticos, fotografias são muito utilizadas, porém novas ferramentas são necessárias para facilitar a visualização dos pacientes quanto aos ganhos estéticos. Os scanners faciais 3D surgem como alternativa para tal finalidade. O objetivo deste estudo foi analisar, através de fotografias e escaneamento 3D da face, o ganho de volume labial após intervenção de preenchimento. A paciente apresentava insatisfação com relação aos seus lábios. Durante a avaliação, constatou-se ausência de volume e contorno labial. Antes de iniciar o procedimento, realizou-se protocolo fotográfico e escaneamento facial (E1). Para o procedimento, foi utilizada uma seringa de 1 mL de ácido hialurônico. O escaneamento foi feito no pós-imediato (E2), após 10 dias (E3) e após 40 dias da intervenção (E4). Nesta mesma consulta após 40 dias, foi realizado injeção da segunda seringa de ácido hialurônico escaneamento do pós-imediato (E5). Após 40 dias, a paciente voltou (E6). Após análise comparativa das malhas, foi possível verificar ganho de 3 mm entre E1 e E2. Na comparação do pré (E1) e após 10 dias (E3), verificou-se volumização total de 1,85 mm, demonstrando que quase 40% do volume no pós-imediato é edema. Em E6, após 40 dias da aplicação da segunda seringa, foi possível quantificar ganho de 2,12 mm no lábio superior quando comparado a E1. Pode-se concluir que o escaneamento 3D contribui expressivamente para avaliação dos resultados alcançados em procedimentos estéticos.
Facial analysis is an important stage in the diagnosis and evaluation of the results obtained in orofacial harmonization. To evaluate the results of aesthetic procedures, photographs are widely used, but new tools are needed to facilitate patient visualization of aesthetic gains. 3D facial scanners have emerged as an alternative for this purpose. The aim of this study was to analyze, by means of photographs and 3D facial scanning, the labial volume gain after a filling intervention. The patient was dissatisfied with her lips. During the evaluation, it was observed the absence of lip volume and contour. Before starting the procedure, a photographic protocol and facial scanning (S1) were performed. A 1 mL syringe of hyaluronic acid was used for the procedure. The scanning was done post-immediately (S2), 10 days (S3) and 40 days after the intervention (S4). In the same appointment after 40 days, the second syringe of hyaluronic acid was injected and scanned post-immediate (S5). After 40 days, the patient returned (E6). After comparative mesh analysis, it was possible to verify a 3 mm gain between S1 and S2. In the comparison of pre (S1) and after 10 days (S3), there was a total volumization of 1.85 mm, showing that almost 40% of the volume in the post-immediate period is edema. In S6, 40 days after the application of the second syringe, it was possible to quantify a gain of 2.12 mm in the upper lip when compared to S1. It can be concluded that 3D scanning contributes expressively to the evaluation of the results achieved in aesthetic procedures.
El análisis facial es una etapa importante en el diagnóstico y la evaluación de los resultados obtenidos en la armonización orofacial. Para evaluar los resultados de los procedimientos estéticos, las fotografías son ampliamente utilizadas, pero se necesitan nuevas herramientas que faciliten al paciente la visualización de las ganancias estéticas. Los escáneres faciales 3D han surgido como una alternativa para este fin. El objetivo de este estudio fue analizar, mediante fotografías y escaneado facial 3D, la ganancia de volumen labial tras una intervención de relleno. La paciente estaba insatisfecha con sus labios. Durante la evaluación, se observó la ausencia de volumen y contorno labial. Antes de iniciar el procedimiento, se realizó un protocolo fotográfico y un escaneado facial (S1). Para el procedimiento se utilizó una jeringa de 1 ml de ácido hialurónico. La exploración se realizó inmediatamente después (S2), a los 10 días (S3) y a los 40 días de la intervención (S4). En la misma consulta, a los 40 días, se realizó la inyección de la segunda jeringa de ácido hialurónico tras la exploración inmediata (S5). A los 40 días, la paciente regresó (E6). Tras el análisis comparativo de las mallas, se pudo comprobar una ganancia de 3 mm entre S1 y S2. En la comparación del pre (S1) y después de 10 días (S3), hubo una volumización total de 1,85 mm, demostrando que casi 40% del volumen post-immediato es edema. En S6, 40 días después de la aplicación de la segunda jeringa, fue posible cuantificar una ganancia de 2,12 mm en el labio superior en comparación con S1. Se puede concluir que el escaneo 3D contribuye expresivamente para la evaluación de los resultados alcanzados en procedimientos estéticos.
RESUMO
Endodontic treatment consists of decontamination of the root canal using irrigants like Sodium Hypochlorite (NaOCl). However, some microorganisms resist conventional therapies, causing relapse and secondary infections. Substances with microbicidal power and potential to become new irrigants need to be studied. Thus, the objective of the present study was to evaluate the antimicrobial potential of Psidium guajava L. (guava tree) extract against clinical and standard (ATCC) strains of Enterococcus faecalis and Candida albicans. For this purpose, the extract was produced from the raw material (leaves and shoots) diluted in the hydroethanolic vehicle (EtOH: H2O / 50:50) for subsequent microbiological analysis. The Minimum Inhibitory Concentration (MIC) and Minimum Microbicidal Concentration (MMC) of the plant extract were tested, according to the Clinical and laboratory standards institute (CLSI) guidelines, on four strains: E. faecalis (ATCC and clinic) and C. albicans (ATCC and clinic). The extract of P. guajava did not produce antifungal action on C. albicans, however, it showed microbicidal potential against strains of E. faecalis, showing MIC of 0.20%. This concentration was lower than the MIC of NaOCl which was 0.31%, a solution that is commonly used in dental clinics. In conclusion, the hydroethanolic extract of P. guajava presents bactericidal action against E. faecalis, being a natural product with potential for future studies regarding the development of new endodontic irrigants.
O tratamento endodôntico consiste na descontaminação do canal radicular com emprego de soluções irrigadoras, como por exemplo, o Hipoclorito de Sódio (NaOCl). Contudo, alguns microrganismos resistem as terapias convencionais, causando recidiva e infecções secundárias. Substâncias com poder microbicida e potencial para se tornarem novas soluções irrigadoras precisam ser estudadas. Assim, o objetivo do presente estudo foi avaliar o potencial antimicrobiano do extrato de Psidium guajava L. (goiabeira) contra cepas clínicas e padrão (ATCC) de Enterococcus faecalis e Candida albicans. Para tanto, o extrato foi produzido a partir da matéria bruta (folhas e brotos) diluído no veículo hidroetanólico (EtOH: H2O / 50:50) para posterior análise microbiológica. Foram testadas a Concentração Inibitória Mínima (CIM) e Concentração Microbicida Miníma (CMM) do extrato vegetal, de acordo com as normas da Clinical and laboratory standards institute (CLSI), sobre quatro cepas: E. faecalis (ATCC e clínica) e C. albicans (ATCC e clínica). O extrato de P. guajava não produziu ação antifúngica sobre C. albicans, contudo, apresentou potencial microbicida contra cepas de E. faecalis, exibindo CIM de 0,20%. Esta concentração foi menor que a CIM do NaOCl que foi de 0,31%, uma solução que é comumente utilizada nas clínicas odontológicas. Em conclusão, o extrato hidroetanólico de P. guajava apresenta ação bactericida contra E. faecalis, sendo um produto natural com potencial para futuros estudos quanto ao desenvolvimento de novos irrigantes endodônticos.
El tratamiento endotónico consiste en la descontaminación del canal raíz con el uso de soluciones de irrigación, como el Hipoclorito de Sodio (NaOCl). Sin embargo, algunos microorganismos se resisten a las terapias convencionales, causando recaídas e infecciones secundarias. Es necesario estudiar las sustancias con poder de microbicida y el potencial para convertirse en nuevas soluciones de riego. Por lo tanto, el objetivo de este estudio fue evaluar el potencial antimicrobiano del extracto de Psidium guajava L. (guava) frente a cepas clínicas y estándar (ATCC) de Enterococcus faecalis y Candida albicans. Con este fin, el extracto se produjo a partir de la materia prima (hojas y brotes) diluida en el vehículo hidroetanólico (EtOH: H2O / 50:50) para análisis microbiológicos posteriores. La Concentración Mínima Inhibidora (CMI) y la Concentración de Microbicidas Minima (CCM) del extracto vegetal se probaron de acuerdo con el Clinical and laboratory standards institute (CLSI) en cuatro cepas: E. faecalis (ATCC y clínico) y C. albicans (ATCC y clínico). El extracto de P. guajava no produjo acción antifúngica sobre C. albicans, sin embargo mostró potencial microbicida frente a cepas de E. faecalis, presentando CIM 0,20%. Esta concentración fue menor que la CMI de NaOCl, que fue del 0,31%, una solución comúnmente utilizada en clínicas dentales. En conclusión, el extracto hidroetanólico de P. guajava muestra una acción bactericida frente a E. faecalis, siendo un producto natural con potencial para futuros estudios sobre el desarrollo de nuevos irrigantes endotónicos.
RESUMO
Microorganisms are capable to combat defense cells by means of strategies that contribute to their stabilization and proliferation in invaded tissues. Frequently antimicrobial-resistant strains appear; therefore, alternative methods to control them must be investigated, for example, the use of plant products. The capacity of the thyme extract (Thymus vulgaris L.) and phytocompound thymol in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in murine macrophages (RAW 264.7) was evaluated. Minimal inhibitory concentrations (MIC) of the plant products were used. The effect of these MIC were analyzed in the assays of phagocytosis and immunoregulation by analysis of the production of cytokines (IL-1ß, TNF-α, and IL-10) and nitric oxide (NO). The plant products effectively assisted the macrophages in the phagocytosis of microorganisms, presenting significant reductions of S. aureus and P. aeruginosa. The macrophages also regulated the production of inflammatory mediators in the infections by S. aureus, P. aeruginosa, and C. albicans. In addition, thyme provided a satisfactory effect in response to the bacterial infections, regarding generation of NO. Thus, the effectiveness of the thyme and thymol to control in vitro infections by S. aureus, P. aeruginosa, and C. albicans was observed. HIGHLIGHTS: Phagocytosis of S. aureus by RAW 264.7 was enhanced with thymol Thyme enhanced the phagocytosis of P. aeruginosa by RAW 264.7 Plant products provided immunoregulation of inflammatory cytokines Production of nitric oxide was improved with the treatments in bacterial infections.
Assuntos
Candida albicans/fisiologia , Candidíase/tratamento farmacológico , Fatores Imunológicos/farmacologia , Macrófagos/imunologia , Extratos Vegetais/farmacologia , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/fisiologia , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/fisiologia , Timol/farmacologia , Animais , Citocinas/metabolismo , Macrófagos/microbiologia , Camundongos , Óxido Nítrico/metabolismo , Fagocitose , Células RAW 264.7 , Thymus (Planta)/imunologiaRESUMO
Os microrganismos resistentes a diferentes classes de agentes antimicrobianos têm se tornado cada vez mais comuns e atualmente são denominados como multirresistentes. Nos hospitais, tais microrganismos apresentam maior perigo, pois são causadores de infecções nosocomiais e a higienização bucal deficiente dos pacientes internados pode tornar a cavidade bucal um sítio para proliferação desses microrganismos multirresistentes. Diante do exposto, novos compostos com ação antimicrobiana precisam ser estudados. O objetivo deste estudo foi avaliar quimicamente o extrato hidroalcóolico de própolis verde de Baccharis dracunculifolia e de Cinnamomum verum (canela) que foram obtidos a partir da extração da matériaprima, analisar a atividade antimicrobiana e antibiofilme dos extratos isolados e combinados contra quatro cepas clínicas multirresistentes de Pseudomonas aeruginosa e Acinetobacter baumannii e verificar a citotoxicidade dos produtos vegetais in vitro em linhagem celular de queratinócitos humanos (HaCat). Para tanto, os extratos vegetais foram preparados a partir da matéria-prima da canela em casca e da própolis bruta. Em seguida, foram caracterizados quimicamente por cromatografia líquida de alta eficiência (HPLC-DAD) para identificação dos principais compostos e a análise do teor de sólidos solúveis dos extratos vegetais também foi realizada. Para avaliação antimicrobiana, foram performados o teste de microdiluição em caldo de acordo com a Clinical and Laboratory Standards Institute (CLSI) e a análise de Checkerboard, para avaliar o efeito combinado dos extratos. A atividade antibiofilme dos extratos combinados foi realizada por meio do teste de MTT, no qual diferentes tempos de contato (5 e 30 min) e diferentes modalidades (inibição na formação do biofilme bacteriano e erradicação do biofilme bacteriano já formado) foram testadas. Para ação citotóxica, as células foram cultivadas em meio DMEM e semeadas na placa de 96 poços. Após aderência inicial, aplicou-se os extratos em diferentes concentrações baseadas nas análises microbiológicas para avaliação da viabilidade celular por meio do teste de MTT. Os dados foram analisados por ANOVA e teste de Tukey, ou Kruskal-Wallis e Dunn, considerando um nível de significância de 5%. Os compostos identificados no extrato de própolis verde de B. dracunculifolia foram ácido clorogênico, derivado do ácido cinâmico e apigenina. O aldeído cinâmico foi o principal composto identificado no extrato de C. verum. Os extratos vegetais apresentaram ação bactericida sobre todas as cepas analisadas e, quando combinados, os extratos atuaram de modo aditivo e algumas combinações sinérgicas foram encontradas. O protocolo de inibição da formação do biofilme promoveu percentuais de redução superiores quando comparado ao protocolo de erradicação. Valores expressivos de 83,86% (p < 0,05) de inibição da formação de biofilme de uma cepa clínica de A. baumannii e 89,31% (p < 0,05) de inibição em uma cepa clínica de P. aeruginosa foram encontrados com a aplicação dos extratos combinados. A atuação dos produtos vegetais foi estatisticamente semelhante a atuação da clorexidina 0,12%. Em conclusão, os extratos de própolis verde e canela na forma isolada ou combinada apresentaram ação antimicrobiana e antibiofilme sobre cepas clínicas de A. baumannii e P. aeruginosa multirresistentes. Dessa forma, os produtos vegetais são promissores agentes antissépticos para futuras formulações odontológicas. (AU)
Microorganisms resistant to different classes of antimicrobial agents have become increasingly common and are currently called multidrug resistant. In hospitals, such microorganisms are more dangerous, as they cause nosocomial infections and poor oral hygiene in hospitalized patients can make the oral cavity a site for the proliferation of these multiresistant microorganisms. Given the above, new compounds with antimicrobial action need to be studied. The objective of this study was to chemically evaluate the hydroalcoholic extract of green propolis from Baccharis dracunculifolia and Cinnamomum verum (cinnamon) that were obtained from the extraction of the raw material, to analyze the antimicrobial and antibiofilm activity of the isolated and combined extracts against four clinical strains multiresistant strains of Pseudomonas aeruginosa and Acinetobacter baumannii and verify the cytotoxicity of plant products in vitro in human keratinocyte cell lineage (HaCat). For this purpose, plant extracts were prepared from raw cinnamon bark and raw propolis. Then, they were chemically characterized by high performance liquid chromatography (HPLC-DAD) to identify the main compounds and the analysis of the soluble solids content of the plant extracts was also performed. For antimicrobial evaluation, the broth microdilution test according to the Clinical and Laboratory Standards Institute (CLSI) and the Checkerboard analysis were performed to evaluate the combined effect of the extracts. The antibiofilm activity of the combined extracts was performed using the MTT test, in which different contact times (5 and 30 min) and different modalities (inhibition of bacterial biofilm formation and eradication of already formed bacterial biofilms) were tested. For cytotoxic action, cells were cultured in DMEM medium and seeded in the 96-well plate. After initial adhesion, the extracts were applied at different concentrations based on microbiological analyzes to assess cell viability through the MTT test. Data were analyzed by ANOVA and Tukey's test, or Kruskal-Wallis and Dunn, considering a significance level of 5%. The compounds identified in the green propolis extract of B. dracunculifolia were chlorogenic acid, cinnamic acid derivative and apigenin. Cinnamic aldehyde was the main compound identified in the C. verum extract. The plant extracts showed bactericidal action on all strains analyzed and, when combined, the extracts acted additively and some synergistic combinations were found. The biofilm formation inhibition protocol promoted higher reduction percentages when compared to the eradication protocol. Significant values of 83.86% (p < 0.05) inhibition of biofilm formation in a clinical strain of A. baumannii and 89.31% (p < 0.05) inhibition in a clinical strain of P. aeruginosa were found with the application of the combined extracts. The performance of plant products was statistically similar to the performance of 0.12% chlorhexidine. In conclusion, extracts of green propolis and cinnamon, in isolated or combined form, showed antimicrobial and antibiofilm action on multiresistant clinical strains of A. baumannii and P. aeruginosa. Thus, plant products are promising antiseptic agents for future dental formulations. (AU)