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.

Efeito de extratos vegetais de Hamamelis virginiana, Persea americana, Cynara scolymus L e Stryphnodendron barbatiman Msobre a expressão fenotípica dos fatores de virulência em biofilmes de Candida albicans / Effect of Hamamelis virginiana, Persea americana, Cynara scolymus L and Stryphnodendron barbatiman M plant extracts on the phenotypic expression of virulence factors in biofilms of the Candida albicans

Objetivo: Analisar a expressão fenotípica de fatores de virulência em biofilmes de Candida albicans frente a extratos glicólicos de plantas. Material e Métodos: Os biofilmes de Candida albicans (ATCC 18804) obtidos a partir de incubação de 48 horas foram expostos por 5 minutos e 24 horas a diferentes concentrações de extratos glicólicos de Hamamelis virginiana e Persea americana, Cynara scolymus L e Stryphnodendron barbatiman M, a fim de verificar a ação antifúngica da proteinase, fosfolipase e hemolisina. Resultados: Todos os extratos foram eficazes na redução do biofilme. Em contato por 5 minutos. os extratos reduziram 50% do biofilme. Após 24 horas. o extrato de Persea americana apresentou o biofilme em 90%, seguido de Cynara scolymus, que o interrompeu em 85%. Houve mudança na intensidade da proteinase após 5 minutos e 24 horas, com uma atividade enzimática média de 0,69 em comparação com o controle de 0,49. Cynara scolymus foi o extrato com maior concentração média de 100 mg/ml; a intensidade da fosfolipase foi alterada com Stryphnodendron barbatiman sendo mais efetivo em 24 horas em relação ao controle (p< 0,0001). A secreção de hemolisina foi modificada por Hamamelis virginiana (12,5 mg/ml) após 5 minutos de exposição e em 24 horas. todos os extratos foram capazes de causar alterações na secreção. Conclusão: Os extratos testados apresentam potencial antifúngico em biofilmes de Candida albicans, implicando em redução significativa dos fatores de virulência. Assim, estes podem ser indicados como uma ferramenta terapêutica alternativa para reduzir a morbidade dessas infecções, já que em ambos os tempos de exposição investigados, eles foram capazes de reduzir a secreção enzimática do fungo (AU)

Objective: Analyze the phenotypic expression of virulence factors in Candida albicans biofilms against plant glycolicextracts. Material and Methods: The biofilms of Candida albicans (ATCC 18804) obtained from incubation for 48 hours were exposed for 5 minutes and 24 hours to different concentrations of glycolic extracts of Hamamelis virginiana and Persea americana, Cynara scolymus L and Stryphnodendron barbatiman M, in order to verify the antifungal activity of the proteinase, phospholipase and hemolysin. Results: All extracts were effective in reducing biofilm. In contact for 5 minutes. the extracts reduced 50% of the biofilm. After 24 hours, the Persea americanaextract showed the biofilm at 90%, followed by Cynara scolymus, which interrupted it at 85%, There was a change in proteinase intensity after 5 minutes and 24 hours. with an average enzymatic activity of 0.69 compared to the control of 0.49. Cynara scolymus was the extract with the highest mean concentration of 100 mg/ml; the phospholipase intensity was changed with Stryphnodendron barbatiman being more effective in 24 hours compared to the control (p< 0.0001). The hemolysin secretion was modified by Hamamelis virginiana (12.5 mg/ml) after 5 minutes of exposure, and in 24 hours. all extracts were capable to cause changes in secretion. Conclusion: The tested extracts have antifungal potential in Candida albicans biofilms, implying a significant reduction in virulence factors. Thus, these can be indicated as an alternative therapeutic tool to reduce the morbidity of these infections, as in both investigated exposure times. they were able to reduce theenzymatic secretion of the fungus (AU)

Antimicrobial action of psidium guajava l. extract against enterococcus faecalis and candida albicans strains: a pilot study / Ação antimicrobiana do extrato de psidium guajava l. contra as cepas de enterococcus faecalis e candida albicans: um estudo piloto / Acción antimicrobiana de la extracción de psidium guajava l. contra los cepes de los enterococcus faecalis y los candida albicans: un estudio piloto

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.

Detoxification of LTA by intracanal medication: analysis by macrophages proinflammatory cytokines production.

The aim of this study was to evaluate in vitro the effect of calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX) on macrophages (RAW 264.7) to produce pro-inflammatory cytokines and nitric oxide after pretreatment with lipoteichoic acid (LTA) of Enterococcus faecalis. Forty-eight human single-rooted teeth were instrumented with R25.08 (RECIPROC) and sterilized by gamma irradiation. LTA was inoculated in the root canal of each specimen for 96 hours. Specimens were instrumented with 40.06 and 50.05 (RECIPROC) and medicated with: I) Pyrogen-free saline solution (SS); II) 2% CHX gel; III) Ca(OH)2 + SS; or IV) Ca(OH)2 + CHX for 14 days. Three samples (S) were performed of the root canal of each specimen at: S1) immediately after instrumentation; S2) after Ethylenediaminetetraacetic acid (EDTA); S3) after intracanal medication removal. Subsequent quantification of cytokines (IL-1ß, TNF-α, MIP-1α, IP-10, G-CSF and IL-6) by immunosorbent assay (ELISA) and nitric oxide by the Griess method was carried-out. Data were submitted to a normality test and then analyzed with one-way ANOVA and Tukey test with a significance level of 5% using GraphPad Prism 6. Ca(OH)2 + SS and Ca(OH)2 + CHX presented lower levels of TNF-α, TNF-α, IL-6, G-CSF and nitric oxide. Ca(OH)2 + SS was the most effective in reducing MIP-1α. CHX was effective in reducing IL-6 and G-CSF. Therefore, the combined intracanal medication of calcium hydroxide and chlorhexidine is effective in reducing the cytokines TNF-α, IL-1ß, IL-6, G-CSF and nitric oxide.

Detoxification of LTA by intracanal medication: analysis by macrophages proinflammatory cytokines production

Abstract The aim of this study was to evaluate in vitro the effect of calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX) on macrophages (RAW 264.7) to produce pro-inflammatory cytokines and nitric oxide after pretreatment with lipoteichoic acid (LTA) of Enterococcus faecalis. Forty-eight human single-rooted teeth were instrumented with R25.08 (RECIPROC) and sterilized by gamma irradiation. LTA was inoculated in the root canal of each specimen for 96 hours. Specimens were instrumented with 40.06 and 50.05 (RECIPROC) and medicated with: I) Pyrogen-free saline solution (SS); II) 2% CHX gel; III) Ca(OH)2 + SS; or IV) Ca(OH)2 + CHX for 14 days. Three samples (S) were performed of the root canal of each specimen at: S1) immediately after instrumentation; S2) after Ethylenediaminetetraacetic acid (EDTA); S3) after intracanal medication removal. Subsequent quantification of cytokines (IL-1β, TNF-α, MIP-1α, IP-10, G-CSF and IL-6) by immunosorbent assay (ELISA) and nitric oxide by the Griess method was carried-out. Data were submitted to a normality test and then analyzed with one-way ANOVA and Tukey test with a significance level of 5% using GraphPad Prism 6. Ca(OH)2 + SS and Ca(OH)2 + CHX presented lower levels of TNF-α, TNF-α, IL-6, G-CSF and nitric oxide. Ca(OH)2 + SS was the most effective in reducing MIP-1α. CHX was effective in reducing IL-6 and G-CSF. Therefore, the combined intracanal medication of calcium hydroxide and chlorhexidine is effective in reducing the cytokines TNF-α, IL-1β, IL-6, G-CSF and nitric oxide.

Resumo O objetivo deste estudo foi avaliar in vitro o efeito do hidróxido de cálcio [Ca(OH)2], clorexidina gel a 2% (CHX) em macrófagos (RAW 264.7) na produção de citocinas pró-inflamatórias e óxido nítrico após pré-tratamento com ácido lipoteicóico (LTA) de Enterococcus faecalis. Quarenta e oito dentes humanos uniradiculares foram instrumentados com R25.08 (RECIPROC) e esterilizados por irradiação gama. LTA foi inoculado no canal radicular de cada espécime por 96 horas. Os espécimes foram instrumentados com 40.06 e 50.05 (RECIPROC) e medicados com: I) solução salina apirogênica (SS); II) CHX 2%; III) Ca(OH)2 + SS; ou IV) Ca(OH)2 + CHX durante 14 dias. Três amostras (S) foram realizadas do canal radicular de cada espécime em: S1) imediatamente após a instrumentação; S2) após ácido etilenodiaminotetracético (EDTA); S3) após a retirada da medicação intracanal. Foi realizada a quantificação subsequente de citocinas (IL-1β, TNF-α, MIP-1α, IP-10, G-CSF e IL-6) por imunoensaio (ELISA) e de óxido nítrico pelo método de Griess. Os dados foram submetidos a um teste de normalidade e então analisados com ANOVA one-way e teste de Tukey com nível de significância de 5% usando GraphPad Prism 6. Ca(OH)2 + SS e Ca(OH)2 + CHX apresentaram níveis mais baixos de TNF -α, TNF-α, IL-6, G-CSF e óxido nítrico. Ca(OH)2 + SS foi o mais eficaz na redução de MIP-1α. CHX foi eficaz na redução de IL-6 e G-CSF. Sendo assim a associação de hidróxido de cálcio e clorexidina é eficaz na redução das citocinas TNF-α, IL-1β, IL-6, G-CSF e óxido nítrico.

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)

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.

The influence of adrenergic blockade in rats with apical periodontitis under chronic stress conditions.

OBJECTIVE: To investigate the influence of chronic stress and adrenergic blockade in a rat model of apical periodontitis. METHODS: Thirty-two Wistar rats were submitted to an animal model of periapical lesion and randomly divided into 4 groups (n = 8): no stress (NS); stress + saline solution (SS); stress + ß-adrenergic blocker (Sß); stress + α-adrenergic blocker (Sα). The SS, Sß and Sα groups were submitted to an animal model of chronic stress for 28 days and received daily injections of saline solution, propranolol (ß adrenergic blocker) and phentolamine (α adrenergic blocker), respectively. After 28 days the animals were euthanized and the following analyses were carried out: a) serum corticosterone levels through Radioimmunoassay; b) measurement of serum levels of IL-1B, IL-6, IL-10 and IL-17 by enzyme-linked immunosorbent assay (ELISA); c) volume of periapical bone resorption by micro-computed tomography; d) histomorphometric analysis by staining with hematoxylin and eosin; e) expression of ß-AR, α-AR, receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) by immunohistochemistry; f) tartrate-resistant acid phosphatase (TRAP) staining; g) ex-vivo cytokine release followed by the stimulation with LPS in superfusion system, by ELISA. RESULTS: SS group displayed significantly higher corticosterone levels than NS group (non-stressed). Higher IL-1ß serum level was observed in the NS group (p < .05); compared to all stressed groups. Other cytokines were present in similar amounts in the serum of all groups. All groups presented similar periapical lesions. All groups presented moderate inflammatory infiltrate, without statistically significant differences between them. No differences were observed regarding ß-AR, α-AR, Rank-L and OPG expression. The number of TRAP-positive cells was significantly decreased in the groups that received daily injections of adrenergic blockers. The IL-1ß release followed LPS stimulation was significantly suppressed when the superfusion media contained propranolol (p < .05). Perfusion containing phentolamine induced a greater release of IL-10. TGF-ß was significantly suppressed by phentolamine perfusion in the NS group (p < .05). CONCLUSIONS: Chronic stress can significantly change the inflammatory cytokines release. Rank-L/OPG system and periapical lesion volume were not affected following the current method applied. The administration of adrenergic blockers was not able to modulate the inflammatory response but presented effectivity in reducing the number of osteoclasts in the periapical region.

Schinus terebinthifolius (Brazilian Peppertree) extract used as antifungal to control Candida spp. in planktonic cultures and biofilms / Extrato de Schinus terebinthifolius (pimenta rosa) utilizado como antifúngico para controlar Candida spp. em culturas planctônicas e biofilmes

Objective: The use of medicinal plants may be an alternative method for the control of Candida spp. Responsible for human infections. This study evaluated the antifungal effect of Schinus terebinthifolius extract (Brazilian Peppertree) on C. albicans, C. dubliniensis, C. glabrata, and C. krusei planktonic cultures and bio films. Material and Methods: Minimum inhibitory concentration (MIC) and minimum fungal concentration (MFC) of the plant extract were determined by the broth microdilution method. Biofilms formed in microplate wells were exposed to the extract for 5 min (50, 100 and 200 mg/mL) or 24 h (25, 50 and 100 mg/mL). After determination of colony-forming units per milliliter (CFU/mL), the data were analyzed by one-way ANOVA and Tukey's Test (P ≤ 0.05). Results: Different MIC (mg/mL) were found, such as 0.39 (C. dubliniensis), 1.56 (C. albicans), and 3.13 (C. glabrata and C. krusei). Besides, MFC (mg/mL) of 0.78 (C. dubliniensis) and 3.13 (C. albicans, C. glabrata and C. krusei) were also observed. Regarding the biofilms, significant reductions (log10) were found after 5 min and 24 h exposure to the plant extract, compared to the control group. However, C. dubliniensis was significantly affected only in 24 h treatment. Conclusion: S. terebinthifolius extract presented a significant antifungal effect on C. albicans, C. dubliniensis, C. glabrata, and C. Krusei both in planktonic cultures and biofilms (AU)

Objetivo: O uso de plantas medicinais pode ser um método alternativo para o controle de Candida spp. responsáveis por infecções humanas. Este estudo avaliou o efeito antifúngico do extrato de Schinus terebinthifolius (pimenta rosa) sobre culturas planctônicas e biofilmes de C. albicans, C. dubliniensis, C. glabrata e C. krusei. Material e Métodos: Concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM) do extrato vegetal foram determinadas pelo método de micro diluição em caldo. Biofilmes formados em poços de microplacas foram expostos ao extrato por 5 min (50, 100 e 200 mg/mL) ou 24 h (25, 50 e 100 mg/mL). Após determinação de unidades formadoras de colônias por mililitro (UFC/mL), os dados foram analisados por one-way ANOVA e Teste de Tukey (P ≤ 0,05). Resultados: Foram encontradas diferentes CIM (mg/mL), como 0,39 (C. dubliniensis),1,56 (C. albicans) e 3,13 (C. glabrata e C. krusei). Além disso, CFM (mg/mL) de 0,78 (C. dubliniensis) e 3,13 (C. albicans, C. glabrata e C. krusei) também foram observadas. Em relação aos biofilmes, foram encontradas reduções significativas (log10) após 5 min e 24 h de exposição ao extrato vegetal, em comparação ao grupo controle. No entanto, C. dubliniensis foi significativamente afetada apenas no tratamento de 24 h. Conclusão: O extrato de S. terebinthifolius apresentou efeito antifúngico significativo sobre C. albicans, C. dubliniensis, C. glabrata e C. Krusei, tanto em culturas planctônicas quanto em biofilmes. (AU)

Microbial Profile and Endotoxin Levels in Primary Periodontal Lesions with Secondary Endodontic Involvement.

This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Microbial profile and endotoxin levels in primary periodontal lesions with secondary endodontic involvement

Abstract This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Resumo Este estudo foi realizado para investigar o perfil microbiano e os níveis de endotoxina de lesões endoperiodontais de origem periodontal. Amostras periodontais e endodônticas foram obtidas de bolsas periodontais e canais radiculares necróticos de 10 dentes com lesões endoperiodontais. A investigação de 40 espécies bacterianas diferentes foram determinadas em cada amostra endodôntica e periodontal usando o método de hibridização de DNA-DNA (checkerboard) e o ensaio cinético cromogênico LAL foi usado para quantificação de endotoxinas. O teste exato de Fisher correlacionou as espécies bacterianas com a microbiota endodôntica ou periodontal. Os níveis de endotoxina (EU/mL) encontrados nas amostras do canal radicular e na bolsa periodontal foram comparados pelo teste de Wilcoxon (p<0,05). Bactérias e unidades de LPS foram encontradas em 100% das amostras endodônticas e periodontais. As espécies E. faecium, P. acnes, G. morbillorum, C. sputigena e L. buccalis foram fortemente correlacionadas com a microbiota endodôntica e P. nigrescens com a microbiota periodontal. P. intermedia, P. endodontalis e V. parvula foram mais prevalentes em ambas microbiotas endodôntica e periodontal. Os níveis de endotoxina na bolsa periodontal (89600 EU/mL) foram significativamente maiores do que no canal radicular (2310 EU/mL). Concluiu-se que a microbiota presente nos tecidos periodontal e endodôntico é semelhante, com maior prevalência de espécies do complexo laranja e maior nível de endotoxina nas bolsas periodontais.

Total protein level reduction of odontopathogens biofilms by Rosmarinus officinalis L. (rosemary) extract: an analysis on Candida albicans and Streptococcus mutans / Redução do nível de proteína total de biofilmes de odontopatógenos por extrato de Rosmarinus officinalis L. (alecrim): uma análise sobre Candida albicans e Streptococcus mutans

Objective: The resistance of fungi and bacteria to the available antimicrobials has increased and the development of alternative products to control them has become a very requirement. The use of plant products could be a viable option due to the efficacy, viability, and availability they present. Thereby, this study evaluated the effect of R. officinalis L. extract on C. albicans and S. mutans biofilms, by the total protein level analysis presented by the microorganisms. Material and Methods: For this purpose, monomicrobial biofilms were formed for 48 h and exposed to the R. officinalis L. extract for 5 min. Then, total protein quantification was performed by Lowry method. Results: The analysis showed significant total protein reduction of the biofilms after exposure to the plant extract, with 39 ± 11%, for C. albicans, and 32 ± 11%, for S. mutans. Conclusion: R. officinalis L. extract decreased the total protein level in both biofilms. Thus, C. albicans and S. mutans protein composition could be a target for action of antimicrobial agents. (AU)

Objetivo: A resistência de fungos e bactérias aos antimicrobianos disponíveis tem se elevado e o desenvolvimento de produtos alternativos para controlar micróbios tem se tornado uma necessidade real. A utilização de produtos de origem vegetal poderia ser uma opção viável, devido à eficácia, viabilidade e disponibilidade que apresentam. Sendo assim, este estudo avaliou o efeito do extrato de R. officinalis L. (alecrim) sobre biofilmes de C. albicans and S. mutans, analisando o nível de proteína total apresentada pelos micro-organismos. Material e Métodos: Para tanto, biofilmes monomicrobianos foram formados por 48 h e expostos ao extrato de R. officinalis L. por 5 min. Então, a quantificação de proteína total foi realizada por método de Lowry. Resultados: A análise demonstrou reduções significativas de proteína total de cada biofilme após exposição ao extrato, sendo 39 ± 11% no biofilme de C. albicans e 32 ± 11%, no caso de S. mutans. Conclusão: O extrato de R. officinalis L. diminuiu o nível de proteína total em ambos os biofilmes. Com isso, a composição proteica de C. albicans e S. mutans poderia ser um alvo para ação de agentes antimicrobianos. (AU)

In vitro and in vivo biological performance of porous Ti alloys prepared by powder metallurgy.

Titanium (Ti) and Ti-6 Aluminium-4 Vanadium alloys are the most common materials in implants composition but ß type alloys are promising biomaterials because they present better mechanical properties. Besides the composition of biomaterial, many factors influence the performance of the biomaterial. For example, porous surface may modify the functional cellular response and accelerate osseointegration. This paper presents in vitro and in vivo evaluations of powder metallurgy-processed porous samples composed by different titanium alloys and pure Ti, aiming to show their potential for biomedical applications. The porous surfaces samples were produced with different designs to in vitro and in vivo tests. Samples were characterized with scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and elastic modulus analyses. Osteogenic cells from newborn rat calvaria were plated on discs of different materials: G1-commercially pure Ti group (CpTi); G2-Ti-6Al-4V alloy; G3-Ti-13 Niobium-13 Zirconium alloy; G4-Ti-35 Niobium alloy; G5-Ti-35 Niobium-7 Zirconium-5 Tantalum alloy. Cell adhesion and viability, total protein content, alkaline phosphatase activity, mineralization nodules and gene expression (alkaline phosphatase, Runx-2, osteocalcin and osteopontin) were assessed. After 2 and 4 weeks of implantation in rabbit tibia, bone ingrowth was analyzed using micro-computed tomography (µCT). EDS analysis confirmed the material production of each group. Metallographic and SEM analysis revealed interconnected pores, with mean pore size of 99,5µm and mean porosity of 42%, without significant difference among the groups (p>0.05). The elastic modulus values did not exhibit difference among the groups (p>0.05). Experimental alloys demonstrated better results than CpTi and Ti-6Al-4V, in gene expression and cytokines analysis, especially in early experimental periods. In conclusion, our data suggests that the experimental alloys can be used for biomedical application since they contributed to excellent cellular behavior and osseointegration besides presenting lower elastic modulus.

Strategies to optimize MEDLINE and EMBASE search strategies for anesthesiology systematic reviews. An experimental study

ABSTRACT BACKGROUND: A high-quality electronic search is essential for ensuring accuracy and comprehensiveness among the records retrieved when conducting systematic reviews. Therefore, we aimed to identify the most efficient method for searching in both MEDLINE (through PubMed) and EMBASE, covering search terms with variant spellings, direct and indirect orders, and associations with MeSH and EMTREE terms (or lack thereof). DESIGN AND SETTING: Experimental study. UNESP, Brazil. METHODS: We selected and analyzed 37 search strategies that had specifically been developed for the field of anesthesiology. These search strategies were adapted in order to cover all potentially relevant search terms, with regard to variant spellings and direct and indirect orders, in the most efficient manner. RESULTS: When the strategies included variant spellings and direct and indirect orders, these adapted versions of the search strategies selected retrieved the same number of search results in MEDLINE (mean of 61.3%) and a higher number in EMBASE (mean of 63.9%) in the sample analyzed. The numbers of results retrieved through the searches analyzed here were not identical with and without associated use of MeSH and EMTREE terms. However, association of these terms from both controlled vocabularies retrieved a larger number of records than did the use of either one of them. CONCLUSIONS: In view of these results, we recommend that the search terms used should include both preferred and non-preferred terms (i.e. variant spellings and direct/indirect order of the same term) and associated MeSH and EMTREE terms, in order to develop highly-sensitive search strategies for systematic reviews.

Strategies to optimize MEDLINE and EMBASE search strategies for anesthesiology systematic reviews. An experimental study.

BACKGROUND: A high-quality electronic search is essential for ensuring accuracy and comprehensiveness among the records retrieved when conducting systematic reviews. Therefore, we aimed to identify the most efficient method for searching in both MEDLINE (through PubMed) and EMBASE, covering search terms with variant spellings, direct and indirect orders, and associations with MeSH and EMTREE terms (or lack thereof). DESIGN AND SETTING: Experimental study. UNESP, Brazil. METHODS: We selected and analyzed 37 search strategies that had specifically been developed for the field of anesthesiology. These search strategies were adapted in order to cover all potentially relevant search terms, with regard to variant spellings and direct and indirect orders, in the most efficient manner. RESULTS: When the strategies included variant spellings and direct and indirect orders, these adapted versions of the search strategies selected retrieved the same number of search results in MEDLINE (mean of 61.3%) and a higher number in EMBASE (mean of 63.9%) in the sample analyzed. The numbers of results retrieved through the searches analyzed here were not identical with and without associated use of MeSH and EMTREE terms. However, association of these terms from both controlled vocabularies retrieved a larger number of records than did the use of either one of them. CONCLUSIONS: In view of these results, we recommend that the search terms used should include both preferred and non-preferred terms (i.e. variant spellings and direct/indirect order of the same term) and associated MeSH and EMTREE terms, in order to develop highly-sensitive search strategies for systematic reviews.

Are Desensitizing Toothpastes Equally Biocompatible and Effective Against Microorganisms?

The aims of this study were evaluate cytotoxicity, genotoxicity, antimicrobial activity of desensitizing toothpastes compared to a common one and the surface roughness of tooth enamel submitted to brushing with these toothpastes. Samples of three desensitizing toothpastes (Colgate Sensitive, Sensodyne and Oral B Sensitive) and common toothpaste (Colgate) were placed in contact with gingival human fibroblasts. Cytotoxicity and genotoxocity were measured by MTT assay and micronucleus test. Antimicrobial activity of the toothpastes extracts against C. albicans, S. mutans and S. aureus were assessed. For surface roughness evaluation, bovine teeth were submitted to 10.000 brushing cycles. The results were analyzed statically using Mann-Whitney U, ANOVA and Z tests (p<0.05). All toothpastes caused cytotoxic effect to the cells (p<0.05), except Colgate Sensitive. The toothpastes did not increase the number of micronuclei compared to the untreated control group. Colgate eliminated all the evaluated microorganisms at lower concentrations compared to Colgate Sensitive and Oral B Sensitive, which were not able to eliminate S. aureus. Sensodyne did not reach the minimum microbicidal concentration. The surface roughness of tooth enamel increased after brushing with Colgate Sensitive and Oral B Sensitive, however the comparison between groups showed no difference on the enamel surface roughness presented by desensitizing toothpastes when compared with the common one (p>0.05). Based on these results, we can conclude that although none toothpaste has induced genotoxicity, Colgate Sensitive was also not cytotoxic. Colgate was the most effective against the microorganisms, and there were no differences on the enamel surface roughness between the groups.

Prevalence and Sensitivity of Bacilli and Pseudomonas in the Newborn's Oral Cavity.

The aim of this study was to isolate Enterobacteria and Pseudomonas from the oral cavity of hospitalized newborns (NB) and determine their prevalence and the sensitivity profile to most commonly used antibiotics for this age group. Samples from the oral cavity of NB from 24 to 48 h age were collected using swabs. The samples were inoculated on MacConkey agar, incubated and the colonies counted and identified. For each strain, the minimum inhibitory concentration (MIC) was determined using agar dilution test. Tests for enterobacteria producing extended spectrumß-lactamases (ESBL) were performed using agar diffusion. Descriptive statistics was used for data analysis. Two of the isolated strains were submitted to the susceptibility test in biofilm. Of the collected samples, 8% presented Enterobacteria (mean of 6,141 CFU/mL) and no Pseudomona species was isolated. Positive samples were from NB in accommodation set or in the NB nursery. Enterobacter was the most prevalent genus and some strains were resistant to ampicillin, gentamicin and cephalothin. No ESBL strain was detected. Microorganisms in biofilms were resistant to all antibiotics, with concentrations four times higher than MIC. The presence of enterobacteria in the oral cavity of newborns, especially some strains resistant to normally used antibiotics, warns to the need for care to avoid the early colonization of this niche and the occurrence of a possible hospital infection in this age group.

Are Desensitizing Toothpastes Equally Biocompatible and Effective Against Microorganisms?

Abstract The aims of this study were evaluate cytotoxicity, genotoxicity, antimicrobial activity of desensitizing toothpastes compared to a common one and the surface roughness of tooth enamel submitted to brushing with these toothpastes. Samples of three desensitizing toothpastes (Colgate Sensitive, Sensodyne and Oral B Sensitive) and common toothpaste (Colgate) were placed in contact with gingival human fibroblasts. Cytotoxicity and genotoxocity were measured by MTT assay and micronucleus test. Antimicrobial activity of the toothpastes extracts against C. albicans, S. mutans and S. aureus were assessed. For surface roughness evaluation, bovine teeth were submitted to 10.000 brushing cycles. The results were analyzed statically using Mann-Whitney U, ANOVA and Z tests (p<0.05). All toothpastes caused cytotoxic effect to the cells (p<0.05), except Colgate Sensitive. The toothpastes did not increase the number of micronuclei compared to the untreated control group. Colgate eliminated all the evaluated microorganisms at lower concentrations compared to Colgate Sensitive and Oral B Sensitive, which were not able to eliminate S. aureus. Sensodyne did not reach the minimum microbicidal concentration. The surface roughness of tooth enamel increased after brushing with Colgate Sensitive and Oral B Sensitive, however the comparison between groups showed no difference on the enamel surface roughness presented by desensitizing toothpastes when compared with the common one (p>0.05). Based on these results, we can conclude that although none toothpaste has induced genotoxicity, Colgate Sensitive was also not cytotoxic. Colgate was the most effective against the microorganisms, and there were no differences on the enamel surface roughness between the groups.

Resumo Os objetivos desse estudo foram avaliar a citotoxicidade, genotoxicidade, atividade antimicrobiana de dentifrícios dessensibilizantes em comparação com um comum e também a rugosidade superficial do esmalte dentário submetido à escovação com esses dentifrícios. Amostras de três dentifrícios dessensibilizantes (Colgate Sensitive, Sensodyne e Oral B Sensitive) e um dentifrício comum (Colgate) foram colocadas em contato com fibroblastos gengivais humanos e a citotoxicidade e genotoxidade foram mensuradas pelo ensaio MTT e teste do micronúcleo. A atividade antimicrobiana dos extratos dos dentifrícios contra C. albicans, S. mutans e S. aureus foi determinada. Para a avaliação da rugosidade superficial, espécimes de dentes bovinos foram submetidas à 10.000 ciclos de escovação. Os resultados foram analisados estatisticamente usando os testes Mann-Whitney U, ANOVA e Teste Z (P<0,05). Todos os dentifrícios causaram efeito citotóxico às células (P<0,05), exceto o Colgate Sensitive. Os dentifrícios não aumentaram o número de micronúcleos em comparação com o grupo não tratado. O Colgate foi capaz de eliminar todos os microorganismos avaliados em concentrações mais baixas em comparação com Colgate Sensitive e Oral B Sensitive, que não foram capazes de eliminar os S. aureus. O Sensodyne não atingiu a concentração microbicida mínima para qualquer microorganismo. A rugosidade superficial do esmalte dentário aumentou após a escovação com Colgate Sensitive e Oral B Sensitive, porém a comparação entre os grupos não mostrou diferença na rugosidade superficial do esmalte apresentada por dentifrícios dessensibilizantes quando comparados ao comum (p>0,05). Com base nesses resultados, podemos concluir que, embora nenhum dentifrício tenha induzido genotoxicidade, o Colgate Sensitive também não foi citotóxico. O Colgate foi o mais eficaz contra os microorganismos, e não houve diferença na rugosidade superficial do esmalte entre os grupos.

Prevalence and Sensitivity of Bacilli and Pseudomonas in the Newborn's Oral Cavity

Abstract The aim of this study was to isolate Enterobacteria and Pseudomonas from the oral cavity of hospitalized newborns (NB) and determine their prevalence and the sensitivity profile to most commonly used antibiotics for this age group. Samples from the oral cavity of NB from 24 to 48 h age were collected using swabs. The samples were inoculated on MacConkey agar, incubated and the colonies counted and identified. For each strain, the minimum inhibitory concentration (MIC) was determined using agar dilution test. Tests for enterobacteria producing extended spectrumβ-lactamases (ESBL) were performed using agar diffusion. Descriptive statistics was used for data analysis. Two of the isolated strains were submitted to the susceptibility test in biofilm. Of the collected samples, 8% presented Enterobacteria (mean of 6,141 CFU/mL) and no Pseudomona species was isolated. Positive samples were from NB in accommodation set or in the NB nursery. Enterobacter was the most prevalent genus and some strains were resistant to ampicillin, gentamicin and cephalothin. No ESBL strain was detected. Microorganisms in biofilms were resistant to all antibiotics, with concentrations four times higher than MIC. The presence of enterobacteria in the oral cavity of newborns, especially some strains resistant to normally used antibiotics, warns to the need for care to avoid the early colonization of this niche and the occurrence of a possible hospital infection in this age group.

Resumo O objetivo foi isolar enterobactérias e Pseudomonas da cavidade oral de recém-nascidos hospitalizados (RN) e determinar a prevalência e o perfil de sensibilidade aos antibióticos mais comumente utilizados para este grupo etário. Foram coletadas amostras da cavidade oral de NB com idade de 24-48 horas, usando swab. As amostras foram inoculadas em ágar MacConkey, incubadas e, as colônias contadas e identificadas. Para cada cepa, a concentração inibitória mínima (CIM) foi determinada utilizando teste de ágar diluição. Testes para enterobactérias produtoras de b-lactamases de espectro estendido (ESBL) foram realizados utilizando difusão em ágar. Estatística descritiva foi utilizada para análise dos resultados. Duas das cepas isoladas foram submetidas ao teste de susceptibilidade em biofilme. Das amostras coletadas, 8% apresentaram enterobactérias (média de 6,141 UFC / ml) e nenhuma espécie de Pseudomonas foi isolada. As amostras positivas foram de RN de alojamento conjunto ou RN de berçário. Enterobacter foi o gênero mais prevalente e algumas cepas foram resistentes à ampicilina gentamicina e cefalotina. Não foi detectada cepa ESBL. Micro-organismos em biofilme foram resistentes a todos os antibióticos, em concentrações quatro vezes superiores ao MIC. A presença de enterobactérias em cavidade oral de recém-nascidos, especialmente algumas cepas resistentes aos antibióticos normalmente utilizados, alerta para a necessidade de cuidados, evitando a colonização precoce deste nicho e a ocorrência de possível infecção nosocomial neste grupo etário.