Gellan-Based Hydrogel as a Drug Delivery System for Caffeic Acid Phenethyl Ester in the Treatment of Oral Candida albicans Infections.

Candida albicans can cause various types of oral infections, mainly associated with denture stomatitis. Conventional therapy has been linked to high recurrence, toxicity, and fungal resistance, necessitating the search for new drugs and delivery systems. In this study, caffeic acid phenethyl ester (CAPE) and gellan gum (GG) were studied as an antifungal agent and carrier system, respectively. First, we observed that different GG formulations (0.6 to 1.0% wt/vol) were able to incorporate and release CAPE, reaching a controlled and prolonged release over 180 min at 1.0% of GG. CAPE-GG formulations exhibited antifungal activity at CAPE concentrations ranging from 128 to >512 µg/mL. Furthermore, CAPE-GG formulations significantly decreased the fungal viability of C. albicans biofilms at short times (12 h), mainly at 1.0% of GG (p < 0.001). C. albicans protease activity was also reduced after 12 h of treatment with CAPE-GG formulations (p < 0.001). Importantly, CAPE was not cytotoxic to human keratinocytes, and CAPE-GG formulations at 1.0% decreased the fungal burden (p = 0.0087) and suppressed inflammation in a rat model of denture stomatitis. Altogether, these results indicate that GG is a promising delivery system for CAPE, showing effective activity against C. albicans and potential to be used in the treatment of denture stomatitis.

Synergistic Antibiofilm Action of Cinnamomum verum and Brazilian Green Propolis Hydroethanolic Extracts against Multidrug-Resistant Strains of Acinetobacter baumannii and Pseudomonas aeruginosa and Their Biocompatibility on Human Keratinocytes.

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.

Antimicrobial action of four herbal plants over mixed-species biofilms of Candida albicans with four different microorganisms.

This study aimed to evaluate the antimicrobial effect of four herbal plants glycolic extracts over mixed-species biofilm composed of Candida albicans (C. albicans) and another pathogenic bacterium as alternative therapy to be investigated. Four plants extract of Pfaffia paniculata roots; Hamamelis virginiana leaf, Stryphnodendron barbatiman tree bark and Gymnema sylvestre stem and leaves were tested over multi-species biofilm of C. albicans (ATCC 18804) and Streptococcus mutans (ATCC 35688), Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 4083) or Pseudomonas aeruginosa (ATCC 15442) for 5 min and 24 h and colony forming units per millilitre was calculated. The data were analysed using Kruskal-Wallis with Dunn's test (p ≤ 0.05). All tested extracts showed antimicrobial action over the mixed-species biofilms after 24 h. Some extracts eliminated totally the biofilms. The glycolic extract of P. paniculata, H. virginiana, S. barbatiman and G. sylvestre are effective over mixed-species biofilms and may be indicated as endodontic irrigant or intracanal medication.

Punica granatum L. Extract Antibiofilm Action against Acinetobacter baumannii Carbapenem-Resistant and Biocompatibility over Human Keratinocytes / Ação Antibiofime de Extrato dePunica granatum L Contra Acinetobacter baumannii Resistente a Carbapnêmicos e Biocompatibilidade em Queratinócitos Humanos

Acinetobacter baumannii is a multi-drug resistant microorganism. The objective of this study was to evaluate the antimicrobial and antibiofilm action of the pomegranate natural extract against eight strains of multidrug resistant Acinetobacter baumanii and to assess the extract cytotoxicity in a culture of Human Keratinocytes (HaCat). Broth microdilution method was used to determine the minimum inhibitory and minimum microbicidal concentrations of the extracts. The extract antibiofilm action was analysed by the MTT colorimetric test. The cytotoxicity evaluation was performed by the MTT colorimetric test, which analysed the mitochondrial cellular action, after contact of the extract for 5 min. The results were statistically analysed by ANOVA and Tukey test with a significance level α≤ 0.05. Punica granatum L. (pomegranate) extract had antimicrobial action on all the 8 clinical strains of Acinetobacter baumannii evaluated. The extract showed a significant reduction in metabolic action in biofilm for all the strains, with results statistically different from growth control (p≤0.05%). P. granatum extract applied for 5 min on human keratinocytes (HaCat) promoted cell viability in all the tested concentrations. The pomegranate extract is effective in reducing the multidrug-resistant clinical strains of Acinetobacter baumanni and is biocompatible. (AU)

Acinetobacter baumannii é um microrganismo multirresistente. O objetivo deste estudo foi avaliar a ação antimicrobiana e antibiofilme do extrato natural de romã contra oito cepas de A. baumanii multirresistente e avaliar a citotoxicidade do extrato em uma cultura de queratinócitos humanos (HaCat). O método de microdiluição em caldo foi utilizado para determinar as concentrações inibitórias mínimas e microbicidas mínimas dos extratos. A ação antibiofilme do extrato foi analisada pelo teste colorimétrico MTT. A avaliação da citotoxicidade foi realizada pelo teste colorimétrico MTT, que analisou a ação celular mitocondrial, após contato do extrato por 5 min. Os resultados foram analisados ​​estatisticamente por ANOVA e teste de Tukey com nível de significância α≤ 0,05. O extrato de Punica granatum L. (romã) apresentou ação antimicrobiana em todas as 8 cepas clínicas avaliadas de A. baumannii. O extrato apresentou redução significativa na ação metabólica no biofilme para todas as linhagens, com resultados estatisticamente diferentes do controle de crescimento (p≤0,05%). O extrato de P. granatum aplicado por 5 min em queratinócitos humanos (HaCat) promoveu viabilidade celular em todas as concentrações testadas. O extrato de romã é eficaz na redução das cepas clínicas multirresistentes de Acinetobacter baumanni e é biocompatível. (AU)

Antimicrobial Action, Genotoxicity, and Morphological Analysis of Three Calcium Silicate-Based Cements.

This study is aimed at evaluating five mineral oxides (5MO), mineral trioxide aggregate repair high plasticity (MTA HP), and mineral trioxide aggregate (MTA) in relation to the antimicrobial action over Porphyromonas gingivalis, Porphyromonas endodontalis, Parvimonas micra, Fusobacterium nucleatum, and Prevotella intermedia; the genotoxicity over mouse macrophage (RAW 264.7) and osteoblast (Mg-63) cultures; and the morphological analysis using scanning electron microscopy (SEM) analysis (50 k and ×100 k). Sodium hypochlorite (NaOCl), calcium hydroxide, and saline solution were used as control groups in the different analysis. All data were submitted to a normality test and then analyzed with one-way ANOVA, Tukey, and Kruskal-Wallis and Dunn tests, considering α ≤ 0.05 significance level. It was found that over P. gingivalis and P. endodontalis, there was no a significant difference between the calcium silicate-based cements (CSC) and the control group of saline solution, and only 5MO was similar to the NaOCl group. However, over P. micra, all groups were effective and showed a statistically significant difference compared to the saline solution group. Conversely, none of the groups were effective over F. nucleatum and P. intermedia, except of the NaOCl group. There was a significant difference between 5MO and MTA groups in comparison with NaOCl and MTA HP over osteoblasts and macrophages after 24 hours. SEM images showed small irregular particles interspersed with some elongated needle-like particles and small irregular particles with some larger particles as well as elongated particles. It was concluded that 5MO, MTA, and MTA HP have effective antimicrobial action over P. micra. However, only 5MO is effective over P. gingivalis and P. endodontalis. Besides, 5MO and MTA are not genotoxic over mouse macrophage (RAW 264.7) and osteoblast (Mg-63) cultures.

Effects of Brazilian green propolis extract on planktonic cells and biofilms of multidrug-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa.

Propolis could represent an alternative therapeutic agent for targeting multidrug-resistant bacteria due to its antimicrobial potential. The effect of Brazilian green propolis (BGP) aqueous extract (AqExt) was evaluated on eight multidrug-resistant clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa, as well as on one reference strain for each bacterial species. The minimum bactericidal concentration (MBC) was determined and optimal concentrations were further evaluated in comparison with 0.12% chlorhexidine. The natural extract was chemically characterized by HPLC-DAD analysis. The MBC values ranged between 3.12 and 27.5 mg ml-1. Analysis of bacterial metabolic activity after treatment for 5 min with BGP-AqExt revealed a strong antimicrobial potential, similar to chlorhexidine. The extract comprised several active compounds including quercetin, gallic acid, caffeic and p-coumaric acid, drupani, galangin, and artepillin C. Altogether, the findings suggest that BGP-AqExt is fast and effective against multidrug-resistant strains of K. pneumoniae and P. aeruginosa in planktonic cultures and biofilms.

Clinical Study of Sodium Hypochlorite, Polymyxin B And Limewater Effect on MMP-3,-8,-9 In Apical Periodontitis.

This study aimed to evaluate sodium hypochlorite (NaOCl), limewater (LW), and Polymyxin B (PMB) as irrigants over MMP-3, MMP-8 and MMP-9. Thirty-three patients with apical periodontitis of single-rooted teeth were treated according to three-experimental groups (n=11): group-1: 2.5% NaOCl was used as irrigant; group-2: 2.5% NaOCl for the first two files and LW: [0.14% Ca(OH)2] for the last two files; group-3: 2.5% NaOCl for the first two files and PMB for the last two files. The association of Ca(OH)2 and CHX was used as an intracanal medication in all groups. Four root canal samplings (S) were collected: S1) immediately after access cavity; S2) after biomechanical preparation; S3) after EDTA application; and S4) after removal of the intracanal medication. After quantification of MMP-3, MMP-8, and MMP-9, the data were analyzed by Friedman and Kruskal-Wallis tests and completed by Dunn test (5%). Regardless the used irrigant, there was no difference in reducing MMP-3 or MMP-8 (P=0,5273, P=0,7048 respectively). However, in reducing MMP-9 (P=0,0246) the NaOCl group was the most effective followed by NaOCl+LW group and NaOCl+PMB group respectively. The intracanal medication [Ca(OH)2 + CHX] with the NaOCl and NaOCl+LW was effective in reducing MMP-8 (P<0,0001, P=0,0025) and MMP-9 (P=0,0007, P=0,0047) respectively, but not for the group of NaOCl+PMB which was not effective in reducing MMP-8 or MMP-9 (P=0,1718, P=0,1953) respectively. NaOCl and NaOCl+LW were effective in reducing MMP-9 levels, and this effectivity could be improved by the use of the intracanal medication [Ca(OH)2 + CHX] in reducing MMP-8 and MMP-9 levels.

Extratos glicólicos de Pfaffia paniculata K E Juglans regia L: avaliação do potencial antimicrobiano, anti-inflamatório, citotóxico e genotóxico / Glycolic extracts of Pfaffia paniculata K and Juglans regia L: evaluation of antimicrobial, anti-inflammatory, cytotoxic and genotoxic potential

A resistência antimicrobiana atingiu proporções alarmantes em todo o mundo, sendo que na Europa mortes causadas por micro-organismos multirresistentes superam os índices de mortalidade da AIDS, tuberculose e a gripe. Assim a fitoterapia desponta no combate a esta problemática, com as diversas atividades biológicas de plantas e seus derivados. Portanto os objetivos do presente estudo foram avaliar a ação antimicrobiana, anti-inflamatória, citotoxicidade, genotoxicidade e constituição fitoquímica dos extratos glicólicos de P. paniculata e J. regia. A ação sobre bactérias anaeróbias (Porphyromonas gingivalis, P. endodontalis, Parvimonas micra e Fusobacterium nucleatum) foi realizada por meio dos testes de microdiluição em caldo (Protocolo M11-A8 - CLSI) e sobre biofilmes monotípicos. Já a ação sobre aeróbios foi realizada sobre 3 cepas de Klebsiella pneumoniae multirresistente, com testes sobre culturas planctônicas (Protocolo M7-A9) e biofilmes; Foi realizada a verificação da atividade antimicrobiana sobre biofilmes heterotípicos de Candida albicans associada a Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans ou Pseudomonas aeruginosa. A citotoxicidade e a genotoxicdade dos extratos foram avaliadas sobre macrófagos de camundongos (RAW 264.7) e queratinócitos humanos (HaCat) pelos testes de MTT e micronúcleos, respectivamente. O potencial antiinflamatório foi verificado dosando os níveis de TNF-⍺, IL-10 e IL-1ß pelo teste de ELISA. Os dados obtiveram distribuição normal sendo a análise estatística realizada por ANOVA e Tukey (p<0,05%). Os extratos de P. paniculata e J. regia promoveram CMM de 50 mg/mL para as anaeróbias. Os biofilmes de P. gingivalis e P. micra foram eliminados com 100 e 200 mg/mL dos extratos (5 min) e com as concentrações de 50 e 100 mg/mL por 24 h; F. nucleatum e P. endodontalis obtiveram reduções variando de 80 a 90%. Os biofilmes heterotípicos de C. albicans e S. mutans obtiveram reduções de até 80% após contato por 5 min. com J. regia e 71% para P. paniculata. Os biofilmes multirresistentes de K. pneumoniae obtiveram reduções na atividade metabólica de até 67,9%. P. paniculata promoveu viabilidade celular variando de 61,1% a 133,8% sobre queratinócitos humanos após 24 h de contato com as concentrações de 12,5 a 0,39 mg/mL, enquanto J. regia obteve 43,9 a 128,4% de viabilidade. Os macrófagos de camundongo obtiveram viabilidade de 18,1 a 101,9% com P. paniculata e 35,4 a 60,6% para J. regia. P. paniculata promoveu a redução nos níveis da citocina pró-inflamatória IL-1ß e aumento nos níveis da citocina antiinflamatória IL-10. Já J. regia promoveu a redução da citocina pró-inflamatória TNF-⍺. Ambos os extratos não promoveram genotoxicidade frente as linhagens celulares. A análise fitoquímica evidenciou a presença de benzofenonas e ácido cafeoilquínico nos extratos de P. paniculata e J. regia, respectivamente. Em conclusão, os extratos de P. paniculata e J. regia demonstraram ação antimicrobiana sobre bactérias aeróbias e anaeróbias e multirresistentes com destaque a eliminação dos biofilmes de P. gingivalis, P. endodontalis, P. micra e K. pneumoniae (multirresistentes). Os extratos demonstraram a ausência de toxicidade e genotoxicidade conforme tempo de aplicação e concentração utilizada, além de possuírem potencial anti-inflamatório(AU)

Antimicrobial resistance has reached alarming proportions worldwide, with deaths in Europe caused by multi-resistant microorganisms exceeding the mortality rates from AIDS, tuberculosis and influenza. Thus phytotherapy emerges in the fight against this problem, with the various biological activities of plants and their derivatives. Therefore, the objectives of the present study were to evaluate the antimicrobial, antiinflammatory, cytotoxicity, genotoxicity and phytochemical constitution of the glycolic extracts of P. paniculata and J. regia. The action on anaerobic bacteria (Porphyromonas gingivalis, P. endodontalis, Parvimonas micra and Fusobacterium nucleatum) was carried out by means of broth microdilution tests (Protocol M11-A8 - CLSI) and on monotypic biofilms. The action on aerobes was performed on 3 strains of multi-resistant Klebsiella pneumoniae, with tests on planktonic cultures (Protocol M7-A9) and biofilms; The verification of antimicrobial activity on heterotypic biofilms of Candida albicans associated with Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans or Pseudomonas aeruginosa was also performed. The cytotoxicity and genotoxicity of the extracts were evaluated on mouse macrophages (RAW 264.7) and human keratinocytes (HaCat) by MTT and micronucleus tests, respectively. The anti-inflammatory potential was assessed by the ELISA test, TNF-⍺, IL-10 and IL-1ß levels were measured. The data obtained a normal distribution and the statistical analysis was performed by ANOVA and Tukey (p <0.05%). The extracts of P. paniculata and J. regia promoted CMM of 50 mg / mL for anaerobes. The biofilms of P. gingivalis and P. micra were eradicated with 100 and 200 mg / mL of the extracts (5 min) and with the concentrations of 50 and 100 mg / mL (24 hours); F. nucleatum and P. endodontalis obtained reductions ranging from 80 to 90%. The heterotypic biofilms of C. albicans and S. mutans obtained reductions of up to 80% after contact for 5 minutes with J. regia and 71% for P. paniculata. The multidrug-resistant strains of K. pneumoniae obtained reductions in metabolic activity of up to 67.9%. The P. paniculata extract promoted cell viability ranging from 61.1% to 133.8% on human keratinocytes after 24 h of contact with concentrations of 12.5 to 0.39 mg / mL, while J. regia obtained 43, 9 to 128.4% viability. Mouse macrophages obtained viability from 18.1 to 101.9% with P. paniculata and 35.4 to 60.6% for J. regia. P. paniculata promoted a reduction in the levels of the pro-inflammatory cytokine IL-1ß and an increase in the levels of the anti-inflammatory cytokine IL-10. J. regia promoted the reduction of the pro-inflammatory cytokine TNF-⍺. Both extracts did not promote genotoxicity against cell lines. Phytochemical analysis showed the presence of benzophenones and caffeoylquinic acid in the extracts of P. paniculata and J. regia, respectively. In conclusion, the extracts of P. paniculata and J. regia demonstrated antimicrobial action on aerobic and anaerobic and multiresistant bacteria, with emphasis on the elimination of the biofilms of P. gingivalis, P. endodontalis and P. micra, as well as the reductions of the biofilms of K. pneumoniae multidrug-resistant. The extracts demonstrated the absence of toxicity and genotoxicity according to the time of application and concentration used, in addition to having anti-inflammatory potential(AU)

Atividade antimicrobiana e citotoxicidade dos extratos glicólicos de Pfaffia paniculata e Juglans regia L / Activity antimicrobial and citotocixity of glycol extracts of Pfaffia paniculata K AND Juglans regia L

O objetivo do trabalho foi investigar se os extratos de Pfaffia paniculata K. e Juglans regia L. possuem ação antifúngica, antibacteriana e toxicidade celular, com testes in vitro. Para os testes antifúngicos foram utilizadas cepas ATCC de Candida spp., e para os testes antibacterianos foram utilizadas cepas ATCC de Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans e Pseudomonas aeruginosa. Para a atividade antimicrobiana primeiramente foram determinados os valores da Concentração Inibitória Mínima (CIM) e da Concentração Microbicida Mínima (CMM) dos extratos pelo método de microdiluição em caldo, segundo Clinical and Laboratory Standards Institute (CLSI). Os micro-organismos que apresentaram CMM foram selecionados para os testes em biofilme, no qual foi preparado em fundo de placa com 96 poços, por 48 h. Após os biofilmes foram tratados por 5 min. utilizando as concentrações de 200, 100 e 50 mg dos extratos. Para mensuração da biomassa foi utilizado o teste de Cristal violeta (CV), e para avaliar a atividade metabólica foi utilizado o teste de MTT. A citotoxicidade foi avaliada sobre fibroblastos gengivais humanos (FMM-1) utilizando os mesmos parâmetros de tratamento utilizados para os testes em biofilmes. Foram avaliadas a viabilidade celular pelos testes de MTT, vermelho neutro e cristal violeta. Os dados obtiveram distribuição normal e foram analisados por ANOVA e teste de Tukey, com significância de 5% (p<0.05%). O extrato de P. paniculata demostrou ação antifúngica em biofilmes, com reduções médias de 29,4 e 42,7% nos testes de CV e MTT. Já a ação antibacteriana foi restrita a S. mutans e P. aeruginosa com reduções médias de 15,7 e 28,6% nos respectivos testes. O extrato de J. regia também demostrou ação antifúngica com redução média de 22,2% na biomassa e 31,4% na atividade metabólica. A ação antimicrobiana ficou restrita a P. aeruginosa com reduções médias de 17,7 e 15,6%, indicados pelos testes de CV e MTT. Quanto a citotoxicidade, a média entre os três testes realizados, indicou que após exposição ao extrato de P. paniculata 58,8% das células continuaram viáveis e para J. regia a viabilidade foi de 65,1%. Conclui-se queo extrato de P. paniculata demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana para P. aeruginosa e S. mutans. As concentrações de 200, 100 e 50 mg do extrato demostraram ser citotóxicas conforme nova diretriz de 11 toxicidade. J. regia demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana sobre P. aeruginosa. Apenas a concentração de 200 mg do extrato se mostrou tóxica a FMM-1(AU)

The aim of this study was to investigate whether extracts of Pfaffia paniculata K. and Juglans regia L. have antifungal, antibacterial and cellular toxicity, with in vitro tests. ATCC strains of Candida spp. Were used for antifungal tests, and ATCC strains of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans and Pseudomonas aeruginosa were used for the antibacterial tests. For the antimicrobial activity, the values of the Minimum Inhibitory Concentration (MIC) and the Minimal Microbicidal Concentration (CMM) of the extracts were determined by the microdilution method in broth, according to Clinical and Laboratory Standards Institute (CLSI). The microorganisms that presented CMM were selected for the biofilm tests, in which it was prepared on a 96-well plate bottom for 48 h. After the biofilms were treated for 5 min. Using the concentrations of 200, 100 and 50 mg of the extracts. To measure the biomass, the Violet Crystal test (CV) was used, and the MTT test was used to evaluate the metabolic activity. Cytotoxicity was assessed on human gingival fibroblasts (FMM-1) using the same treatment parameters used for biofilm tests. Cell viability was evaluated by the MTT, neutral red and violet crystal tests. The data obtained normal distribution and were analyzed by ANOVA and Tukey test, with significance of 5%. The extract of P. paniculata showed antifungal action in biofilms, with average reductions of 29.4 and 42.7% in CV and MTT tests; The antibacterial action was restricted to S. mutans and P. aeruginosa with mean reductions of 15.7 and 28.6% in the respective tests. The extract of J. regia also demonstrated antifungal action with an average reduction of 22.2% in biomass and 31.4% in metabolic activity. The antimicrobial action was restricted to P. aeruginosa with mean reductions of 17.7 and 15.6%, indicated by CV and MTT tests. As for cytotoxicity, the mean of the three tests carried out indicated that after exposure to P. paniculata extract 58.8% of the cells remained viable and for viability the viability was 65.1%. In conclusion the extract of P. paniculata showed antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action for P. aeruginosa and S. mutans. The concentrations of 200, 100 and 50 mg of the extract proved to be cytotoxic according to the new toxicity guideline. J. regia demonstrated antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action on P. aeruginosa. Only the 200 mg concentration of the extract was shown to be toxic to FMM-1 (AU)