Down-regulation of complement genes in lipopolysaccharide-challenged zebrafish (Danio rerio) larvae exposed to Indonesian propolis / Regulação negativa dos genes do complemento em larvas de peixe-zebra (Danio rerio) desafiadas com lipopolissacarídeo expostas à própolis indonésia

Abstract Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 μg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 μg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 μg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.

Resumo Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 μg/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 μg/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 μg/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).

Efeito do extrato da própolis e do digluconato de clorexidina sobre a formação de biofilme por Candida albicans em resina acrílica / Effect of propolis extract and chlorhexidine digluconate on the biofilm formation by Candida albicans in acrylic resin

Objetivo: avaliar in vitro a ação antimicrobiana do extrato da própolis (EP) e do digluconato de clorexidina (DCHX) na formação de biofilme por Candida albicans em resina acrílica termopolimerizada. Métodos: o efeito do EP e DCHX em biofilmes de C. albicans foi avaliado pela quantificação de Unidades Formadoras de Colônias (UFCs), pela quantificação da biomassa por cristal violeta e de polissacarídeos por safranina. Para tanto, C. albicans foram pré-aderidas em corpos de prova e somente em microplacas de poliestireno, posteriormente foi realizado o tratamento com diferentes concentrações de EP (221 µg/mL e 443 µg/mL) e DCHX (0,25% e 0,5%). Resultados: foi demonstrada uma redução significativa na formação de biofilme por C. albicans com ambas as substâncias testadas e em todas as concentrações. Conclusão: de acordo com os resultados, observou-se que ambos os tratamentos foram eficazes na redução do biofilme e que o EP, por ser um produto natural, de baixo custo e sem afeitos colaterais, representa uma alternativa inovadora para o tratamento da candidose oral em usuários de próteses removíveis.(AU)

Objective: to evaluate in vitro the antimicrobial action of propolis extract (EP) and chlorhexidine digluconate (DCHX) in the biofilm formation by Candida albicans in thermopolymerized acrylic resin. Methods: the effect of EP and DCHX on C. albicans biofilms were evaluated through the quantification of Colony Forming Units (CFU's), the quantification of biomass by violet crystal and polysaccharides by safranin. For this purpose, C. albicans were pre-adhered to specimens and only on polystyrene microplates and subsequently the treatment was performed with different concentrations of EP (221 µg/mL and 443 µg/mL) and DCHX (0.25% and 0,5%). Results: a significant reduction in the biofilm formation by C. albicans was demonstrated, with both substances tested and in all concentrations. Conclusion: according to the results, it was observed that both treatments were effective in reducing biofilm and EP, being a natural product, low cost and without side effects, represents an innovative alternative for the treatment of oral candidiasis in users removable dentures.(AU)

Atividade antimicrobiana da própolis verde frente a bactérias Gram positivas e Gram negativas resistentes a antimicrobianos comerciais / Antimicrobial activity of green propolis against positive Gram bacteria and Gram negative resistant commercial antimicrobials

Este estudo teve por objetivo avaliar a atividade antimicrobiana do extrato da própolis verde frente a cepas Gram positivas e Gram negativas resistentes a antimicrobianos comerciais. Foi realizado teste de suscetibilidade antimicrobiana das cepas de Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa e Listeria monocytogenes frente a oito antimicrobianos comerciais. Posteriormente foi verificado a atividade antimicrobiana da própolis verde com base na concentração inibitória mínima e concentração bactericida apenas para as cepas que se mostraram resistentes. Foi possível verificar, com exceção de E. coli, que as demais bactérias apresentaram resistência a mais de um antimicrobiano, e o extrato de própolis verde apresentou valores de CIM e CBM variando de 0,18 a 6,20 mg.mL-1 e 0,37 a 50,0 mg.mL-1,respectivamente. O extrato da própolis verde apresenta potencial atividade antimicrobiana em substituição ao uso de antimicrobianos sintéticos.

Antibacterial and Cytotoxic Potential of a Brazilian Red Propolis

Objective: To evaluate in vitro the effect of a red propolis ethanolic extract (RPE) in the prevention of growth of a cariogenic biofilm and its cytotoxic potential. Material and Methods: Minimum inhibitory and bactericidal concentrations (MIC and MBC) of RPE against Streptococcus mutans and Lactobacillus casei were determined. The cytotoxic potential of 0.4% RPE in oral fibroblasts was observed after 1, 3 and 5 min of contact. Cellulose membrane disks (13 mm, N=12) were used for biofilm formation (24 h) of S. mutans and L. casei, which were treated (1 min) with 0.4% RPE or 0.12% Chlorhexidine (CHX). The control group of biofilm formation was not submitted to any treatment. Serial dilutions were then made to evaluate microbial viability. Descriptive data analysis and, for microbial viability, Mann Whitney test were performed (p≤0.05). Results: RPE showed similar MIC and MBC (4.46 mg/mL) against S. mutans and, for L. casei, they were 8.92 mg/mL (MIC) and 17.85 mg/mL (MBC). CHX presented MIC and MBC <0.00002 mg/mL for S. mutans and 0.00047 mg/mL for L. casei. After 1, 3 and 5 min, the RPE exhibited, respectively, 69.38%, 43.91% and 40.36% of viable cells. The RPE (6.55) and CHX (6.87) presented similar efficacy to reduce the total number of viable bacteria (p>0.05). Regarding the total number of viable bacteria (Log10 CFU/mL), the RPE (6.55) and CHX (6.87) presented similar efficacy (p>0.05). Conclusion: Red propolis extract showed antibacterial activity against the tested strains, exhibited acceptable cytotoxicity and reduced the colonization of S. mutans and L. casei in a biofilm membrane model.

Effects of MTA and Brazilian propolis on the biological properties of dental pulp cells

Abstract: The aim of this study was to evaluate the effect of mineral trioxide aggregate (MTA) and Brazilian propolis on the cell viability, mineralization, anti-inflammatory ability, and migration of human dental pulp cells (hDPCs). The cell viability was evaluated with CCK-8 kit after 1, 5, 7, and 9 days. The deposition of calcified matrix and the expression of osteogenesis-related genes were evaluated by Alizarin Red staining and real-time PCR after incubation in osteogenic medium for 21 days. The expression of inflammation-related genes in cells was determined after exposure to 1 μg/mL LPS for 3 h. Finally, the numbers of cells that migrated through the permeable membranes were compared during 15 h. Propolis and MTA significantly increased the viability of hDPCscompared to the control group on days 7 and 9. In the propolis group, significant enhancement of osteogenic potential and suppressed expression of IL-1β and IL-6 was observed after LPS exposure compared to the MTA and control groups. The number of migration cells in the propolis group was similar to that of the control group, while MTA significantly promoted cell migration. Propolis showed comparable cell viability to that of MTA and exhibited significantly higher anti-inflammatory and mineralization promotion effects on hDPCs.

Gum arabic and red propolis protecteting colorectal preneoplastic lesions in a rat model of azoxymethane

Abstract Purpose: To evaluate red propolis, gum arabic and L-lysine activity on colorectal preneoplastic lesions induced by azoxymethane (AOM). Methods: The study featured 4 control groups (I-IV) and 4 experimental groups (V-VIII), totaling 48 rats. Once a week for 2 weeks, animals on control groups received saline, while animals in experimental groups received azoxymethane (15 mg/kg i.p.). The follow up along 16 weeks included daily oral gavage to administer water (I and V), L-lysine (150 mg/kg)(II and VI), própolis (100mg/5ml/kg)(III and VII), or gum arabic (5ml/kg)(IV and VIII). Was performed surgery on the animals in the end of this time in order to collect blood for biological assays (TBARS, GSH), followed by their sacrifice to tissue extract. Results: Oxidative stress (TBARS) and the number of aberrant crypt foci (ACF) in distal colon were lower using própolis (p<0.01 for both parameters). Gum arabic reduced preneoplastic lesions (ACF ≤ 4 crypts) on distal colon and on the entire colon (p<0.05). Conclusions: Red propolis reduced AOM-induced oxidative stress (TBARS) and total number of ACF in the distal colon. L-lysine neither protected against nor enhanced AOM-induced ACF. Gum arabic reduced the number of ACF.

Evaluación in vitro de la actividad antimicrobiana de un propóleo chileno sobre muestras clínicas de exudados bucofaríngeos y cepas ATCC / In vitro evaluation of the antimicrobial activity of a Chilean propolis on clinical samples of oropharyngeal exudates

Propolis is a substance manufactured by Apis mellifera and has been widely used in folk medicine due to its high concentration of bioactive compounds. The purpose of the following study was to characterize and evaluate in vitro the antimicrobial properties of propolis on clinical samples and ATCC strains. The chemical characterization of propolis presents a concentration of total polyphenols of 247 ± 9 mg EAG g-1 MS, flavones and flavonols 75± 4 mg EQ g-1 MS, flavanonones and flavanonols 118 ± 11 EP g-1 MS. HPLC-DAD identified apigenin, galangin, phenethyl ester of caffeic acid and pinocembrin, in addition to 16 compounds by HPLC MS/MS. Chilean propolis is a natural antimicrobial, showing effectiveness in strains ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum and clinical samples of Staphylococcus aureus unlike Escherichia coli. These results demonstrate the antimicrobial effectiveness of the synergy of compounds present in propolis against different human pathogens.

El propóleos es una substancia fabricada por Apis mellifera y ha sido utilizado ampliamente en la medicina popular debido a su alta concentración de compuestos bioactivos. El propósito del siguiente estudio fue caracterizar y evaluar in vitro las propiedades antimicrobianas del propóleos sobre muestras clínicas y cepas ATCC. La caracterización química de propóleos presenta una concentración de polifenoles totales de 247 ± 9 mg EAG g-1 de MS, flavonas y flavonoles 75 ± 4 mg EQ g-1 de MS, flavanononas y flavanonoles 118 ± 11 EP g-1 de MS. Mediante HPLC-DAD se identificó apigenina, galangina, fenetil éster del ácido cafeico y pinocembrina, además de 16 compuestos mediante HPLC MS/MS. El propóleos chileno es un antimicrobiano natural, observándose efectividad en cepas ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum y muestras clínicas de Staphylococcus aureus a diferencia de Escherichia coli. Estos resultados demuestran la efectividad antimicrobiana de la sinergia de compuestos presentes en el propóleos ante diferentes patógenos humanos.

Evaluación de la actividad anti-inflamatoria de propóleos chileno sobre cortes histológicos de orejas de ratón / Evaluation of the anti-inflammatory activity of Chilean propolis on histological sections of mouse ears

RESUMEN: El propóleos es un producto resinoso complejo producido por las abejas Apis mellifera, el cual posee diversas actividades biológicas como inmunomodulador, antiinflamatorio, anticancerígeno, antiviral, antibacteriano, antioxidante, entre otros. El propósito del siguiente estudio fue realizar una evaluación in vivo de las propiedades antiinflamatorias de un extracto de propóleos chileno, sobre el modelo de edema auricular inducido por 13-acetato-12-O-tetradecanoilforbol (TPA) en pabellón auricular de ratón, para posterior evaluación y análisis histológico. El extracto de propóleos chileno (EEP) utilizado se obtuvo a partir de un macerado etanólico, rotaevaporado y liofilizado. Se observó que el EEP disminuyó el edema y el infiltrado inflamatorio de forma significativa. Estos resultados sugieren que el extracto etanólico de propóleos chileno posee potenciales efectos antiinflamatorios o moduladores del sistema inmunológico en edema auricular.

SUMMARY: Propolis is a complex resinous product produced by bees Apis mellifera, which has a number of biological activities such as an immunomodulator, anti-inflammatory, anticarcinogenic, antiviral, antibacterial, antioxidant, among others. The purpose of the following study was to perform an in vivo evaluation of the anti-inflammatory properties of a Chilean propolis extract, on the model of atrial edema induced 12-O-tetradecanoyl phorbol-13- acetate (TPA) in the mouse auricular pavilion, for later evaluation and histological analysis. The Chilean propolis extract (EPP) used was obtained from an ethanolic, rotaevaporated and lyophilized macerate. It was observed that the EPP significantly decreased edema and inflammatory infiltrate. These results suggest that the ethanolic extract of Chilean propolis possesses potential anti-inflammatory or modulatory effects of the immune system in atrial edema.

The anti-caries activity and toxicity of an experimental propolis-containing varnish

Abstract We investigated the anti-caries effects of an experimental propolis varnish in vivo, and further tested its toxicity against fibroblasts. Fifty-six SPF female Wistar rats were infected with Streptococcus mutans UA159 (SM) and allocated into four groups (n = 14/group): G1, propolis varnish (15%/PV); G2, chitosan varnish (CV/vehicle); G3, gold standard (GS/Duraphat®); and G4, untreated. The animals received a single varnish application on their molars and were submitted to a high cariogenic challenge (Diet-2000, 56% sucrose, and 5% sucrose-added water, ad libitum) for 4 weeks. Total cultivable microbiota and SM were counted, and smooth-surface and sulcal caries were scored. PV, CV and GS cytotoxic effects were tested against fibroblasts. The data were analyzed using ANOVA with the Tukey-Kramer test (p ≤ 0.05). Total microbiota and SM counts did not differ among the treatments (p = 0.78), or in relation to the untreated group (p = 0.52). PV reduced development of smooth-surface enamel caries compared with the untreated group (p = 0.0018), with no significant difference from GS (p = 0.92); however, the PV effects were no longer observed when the dentin was affected. Neither PV nor GS prevented enamel sulcal lesion onset, but GS significantly reduced the severity of dentinal sulcal lesions (p < 0.0001). No significant difference was observed in fibroblast viability between PV and GS (p < 0.0001). In conclusion, PV prevented smooth-surface enamel caries and showed low cell toxicity. Nevertheless, due to the high cariogenic challenge, its effects were not sustained throughout the experiment. Further studies are encouraged to establish a protocol to sustain the long-term anti-caries activity of PV in the oral cavity.

Pythium insidiosum: inhibitory effects of propolis and geopropolis on hyphal growth

Abstract Propolis and geopropolis are resinous products of bees showing antimicrobial effects. There is no data concerning their action against Pythium insidiosum - the causative agent of pythiosis, a pyogranulomatous disease of the subcutaneous tissue that affects mostly horses, dogs and humans. Fragments of 15 isolates of P. insidiodum were incubated with propolis and geopropolis extracts and evaluated for up to seven days to detect the minimal fungicidal concentration (MFC). Propolis inhibited three isolates at 1.0 mg mL-1 after 24 h and all other isolates at 3.4 mg mL-1. Geopropolis led to more variable results, exerting predominantly a fungistatic action than a fungicidal one. Propolis was more efficient than geopropolis in inhibiting P. insidiosum since lower concentrations led to no growth after 24 h. This effect may be due to propolis chemical composition, which has more active compounds than geopropolis. Propolis seemed to be a good candidate for in vivo studies, since treatment with conventional antifungal compounds is difficult in most of the cases, requiring extensive surgical debridement.

Propolis protective role against morphological, hormonal biochemical and histological alterations induced by sildenafil overdoses / Rol protector del propóleo contra las alteraciones morfológicas, hormonales, bioquímicas e histológicas inducidas por sobredosis de sildenafil

Sildenafil is widely used for the treatment of erectile dysfunction with few studies are available on the protective role of propolis against its reproductive toxicity. The present study aims to investigate the hormonal biochemical and histomorphometric alterations induced in the testicular tissues by sildenafil overdoses. Four groups of rabbits were exposed to sildenafil with or without propolis as follows: Group I received the formulated vehicle, Group II received sildenafil (3 mg/kg), Group III received propolis (50 mg/kg), Group IV received sildenafil plus propolis. Sildenafil lowered body weight gain, testosterone and follicular stimulating hormone concentration but increased testis index while luteinizing hormone was almost not affected. Moreover, sildenafil treated rabbits showed degenerative seminiferous tubules and disturbance of spermatogenesis together with spermatocytes sloughing and nuclear alterations. Exposure to sildenafil plus propolis ameliorated tubular alterations, spermatogenesis disturbances, hormonal levels changes and partially protected spermatocytes from morphological nuclear alterations but could not ameliorate the effect on the body weight gain and testis index. The findings of the present work may indicate that propolis can ameliorate partially the reproductive toxicity induced by sildenafil overdoses with more need for further studies on the adverse effect of these doses on the other vital organs.

El sildenafil es un medicamento ampliamente utilizado para el tratamiento de la disfunción eréctil y existen pocos estudios disponibles referente a la función protectora del propóleo contra su toxicidad reproductiva. El objetivo fue investigar las alteraciones hormonales, bioquímicas e histomorfométricas, inducidas en los tejidos testiculares por sobredosis de sildenafil. Cuatro grupos de conejos fueron expuestos a sildenafil con o sin propóleo de la siguiente manera: grupo I recibió el sildenafil formulado, grupo II recibió sildenafil (3 mg/kg), grupo III recibió propóleo (50 mg/kg) y el grupo IV recibió sildenafil más propóleo. El sildenafil redujo el peso corporal, la testosterona y la concentración de la hormona foliculoestimulante, sin embargo, se observó un aumento del índice testicular mientras que la hormona luteinizante casi no se vio afectada. Por otra parte, los conejos tratados con sildenafil mostraron degeneración de los túbulos seminíferos, trastornos de la espermatogénesis y alteraciones nucleares de los espermatocitos. Con el uso de sildenafil más propóleo fue posible disminuir las alteraciones de los túbulos seminíferos, los trastornos de la espermatogénesis y los niveles de cambios hormonales; los espermatocitos fueron protegidos parcialmente de alteraciones nucleares morfológicas, pero no pudo mejorar el efecto de aumento de peso corporal e índice testicular. Los resultados indican que el propóleo puede aliviar, en parte, la toxicidad en la reproducción inducida por sobredosis de sildenafil. No obstante, existe la necesidad de realizar más estudios sobre los efectos adversos de estas dosis en otros órganos vitales.

Tissue characterization following revascularization of immature dog teeth using different disinfection pastes

Abstract Revascularization of immature teeth with necrotic pulps traditionally involves the use of triple antibiotic paste, which may sometimes lead to undesirable complications. The objective of this study was to assess tissue repair in immature dog teeth with apical periodontitis subjected to revascularization, comparing two different pastes used for root canal disinfection. Apical periodontitis was induced in 30 dog premolars. Teeth were randomly divided into three experimental groups: root canals filled with triple antibiotic paste (n = 10); root canals filled with 1% propolis paste (n = 10); and no medication (n = 10). An additional group (n = 10, no intervention) was used as control. After 7 months, the jaws were histologically evaluated for the following variables: newly formed mineralized tissue (present/absent); vital tissue in the canal space (absent/periodontal ligament-like/pulp-like); apical extension of root (present/absent); and severity of inflammatory process (absent/mild/moderate/severe). There were no statistically significant differences among the experimental groups in new mineralized tissue formation and apical root development. The formation of vital tissue in the canal space, in turn, was statistically different between the triple paste and propolis groups: vital tissues were present in all revascularized teeth disinfected with propolis paste (100%), compared to 71% of those disinfected with the triple paste. Severity of inflammatory process was different between the triple paste and no medication groups. The new tissues formed onto canal walls and in the root canal space showed characteristics of cementum and periodontal ligament, respectively. Propolis may have some advantages over the triple paste for the revascularization of immature teeth.

Utilización de propóleos en odontología / The use of propolis in dentistry

El propóleos es un producto natural elaborado por las abejas a partir de la secreción que recogen de ciertas especies vegetales y que, luego de modificarlas con sus secreciones salivares, lo transportan al interior de la colmena. Así, el propóleos es responsable directo de garantizar la asepsia de la colmena. Múltiples investigaciones científicas atribuyeronal propóleos propiedades antioxidantes, antibacterianas, antivirales, fungicidas, cicatrizantes, antiinflamatorias, anestésicas, inmunomoduladoras antitumorales. Asimismo, en bastas investigaciones se comprobó que el propóleos actúa inhibiendo la actividad de los Streptococo mutans, principal microorganismo productor de caries dental. Esto motivó la realización de la presente revisión bibliográfica sobre las propiedades y utilización del propóleos en odontología.

Propolis is a natural product made by bees fromcollecting secretion of certain plant speciesand, after modifying their salivary secretions,transported into the hive. So propolis is directlyresponsible for ensuring the cleanliness of the hive.Multiple scientific research attributed to propolisantioxidant, antibacterial, antiviral, fungicide, healing,anti-inflammatory, anesthetic,immunomodulatory and antitumor properties.Also in rough investigations it was foundthat propolis acts by inhibiting the activityof Streptococcus mutans, the main producingmicroorganism tooth decay. This led to therealization of this literature review on theproperties and use of propolis in dentistry.

Anticancer potential of bee venom and propolis combined treatment on human breast adenocarcinoma cell line [MCF-7]

Backgrounds: Natural remedies were used for cancer treatments, particular breast cancer. Also, the consumption of food products containing high amount of flavonoids and antioxidants had reported to lower the risk of various cancers. Bee venom [BV] and propolis were produced by honey bee. They were characterized by naturopathic formulation, affordability and containing high amount of antioxidants. Moreover, they were used safely since ancient times globally. Although that, there is no information about the synergistic or antagonistic anticancer effects of their combination. This study was designed to evaluate cytotoxic and pro-apoptotic effects of BV, propolis, and their combination on breast cancer [MCF-7] cells

Materials and Methods: As preliminary study, MCF-7 cells were treated with BV [5, 10, and 20micro g/ml] and propolis [50, 150, and 450micro g/ml] to specify the desired combination doses of each treatment with no anticancer effect individually. Consequently, doses of [5micro g/ml BV+ 50micro g/ml propolis and 5micro g/ml BV+ 150micro g/ml propolis] were chosen to evaluate the possible synergistic anticancer potential between them. All groups in this study were examined at 2, 4, and 12 hours intervals. The morphological changes were evaluated by acridine orange/ ethidium bromide dual fluorescent staining and Giemsa staining to reveal the formation of apoptotic bodies or nuclear condensation and cytoplasmic blebbing, respectively. DNA fragmentation assay was also carried out to record the reduction in DNA content and apoptosis. Bcl-2 expression, cytoplasmic anti-apoptotic marker, was used to prove the apoptotic properties, and autophagic cell death by florescent microscopy was evaluated also

Results: Morphological observation by inverted and florescent microscopy revealed apoptotic cell death under exposure to BV [10 and 20micro g/ml] and propolis [450micro g/ml]. On the other hand, the results of combined treatments revealed significant morphological alterations after fluorescent and Giemsa staining. Apoptotic DNA fragmentation was clearly observed and Bcl-2 recoded significant down regulation which proved the apoptotic properties of combined treatments. Additionally, autophagic degradation results also supported the occurrence of stress on treated cells leading finally to cell death. All results of powerful anticancer potential were obvious among all combined-treated groups in dose and time dependent manner. This clear that, the combined treatments have possible synergistic effect which, propose it as potential candidates to be used in development of chemotherapy

Actividad antibacteriana in vitro de propóleos chilenos sobre Helicobacter pylori / In vitro antibacterial activity of Chilean propolis against Helicobacter pylori

Introduction: Propolis is a natural product derived from beekeeping. It has anesthetic, anti-inflammatory, immune-stimulant and antibacterial properties on grampositive and gramnegative bacteria. However, little is known regarding its activity on Helicobacter pylori. This bacteria colonizes about half of the world’s population and is associated with chronic gastritis, peptic ulcer and gastric cancer. Objective: The aim of this study was to evaluate the inhibitory activity of 22 propolis extracts from nine of the 11 beekeeping Chilean regions on 10 strains of H. pylori isolated from gastric mucosa. Methods: The antibacterial activity of the extracts was determined using the well diffusion method and diffusion disks. Results: 100% of the extracts were active on the tested strains, showing inhibition halos equal to or greater than 15 mm by both methods. Conclusions: our results show an effective anti H. pylori activity of propolis. However, additional microbiological studies are needed before a potential clinical utility of these natural products is warranted.

Introducción: El propóleos es un producto natural derivado de la apicultura que tiene propiedades anestésicas, anti-inflamatorias, inmuno-estimulantes y antibacterianas. Ejerce su acción sobre distintas bacterias grampositivas y gramnegativas. Sin embargo, es muy poco lo que se sabe en relación a su actividad sobre H. pylori, bacteria asociada con gastritis crónica, úlcera gastro-duodenal y cáncer gástrico y que coloniza a alrededor de la mitad de la población mundial. Objetivo: Evaluar la actividad inhibitoria de 22 extractos de propóleos de orígenes botánicos diferentes, provenientes de nueve de las once zonas mielíferas de Chile, en la época de otoño, sobre 10 cepas de H. pylori aisladas de mucosa gástrica. Metodología: La actividad antibacteriana de los extractos se determinó a través del método de difusión en pocillos y de difusión en discos. Resultados: 100% de los extractos fueron activos sobre las cepas ensayadas, observándose halos de inhibición iguales o mayores a 15 mm en ambos métodos. Conclusiones: Los datos obtenidos in vitro en el presente estudio muestran una efectiva actividad anti H. pylori de los propóleos chilenos, siendo necesario estudios microbiológicos y farmacológicos adicionales para avanzar en una posible utilidad clínica de estos productos naturales.

Higienização bucal com digluconato de clorexidina e extrato etanólico de própolis em pacientes de Unidade de Terapia Intensiva (UTI) de um hospital público na cidade de São Paulo - Brasil / Oral higiene with chlordexdine digluconate and ethanol extract of propolis in patients in the Intensive Care Unit (ICU) of public hospital in São Paulo - Brazil

Focos de infecção na boca têm sido relacionados com o comprometimento da saúde do corpo do indivíduo, despertando o interesse de médicos e dentistas. A infecção é uma complicação frequente e de elevada mortalidade nos pacientes internados em Unidade de Terapia Intensiva. Estes pacientes na maioria das vezes, não possuem higienização bucal adequada, possivelmente pelo desconhecimento de técnicas adequadas pelas equipes de terapia intensiva, e pela ausência do relacionamento odontologia e enfermagem. Pesquisas com produtos naturais visam o tratamento efetivo destas infecções. Na odontologia, tem-se estudado a atividade farmacológica do extrato de própolis em algumas situações, como: gengivites, periodontites, aftas, mumificação pulpar. Também, tem sido usado em curativos pré e pós-cirúrgicos e em tratamentos da candidíase, herpes labial e higiene bucal, devido à capacidade antisséptica e cicatrizante em indivíduos internados em hospitais. O objetivo deste trabalho foi estudar a higienização bucal com água filtrada, digluconato de clorexidina a 0,12% e extrato etanólico de própolis a 6% em pacientes internados na UTI e a atividade do digluconato de clorexidina a 0,12% e do extrato etanólico de própolis a 6% sobres leveduras e em doses subinibitórias sobre a produção de exoenzimas proteinase e fosfolipase e as características fenotípicas (franjas). Foram estudados 150 pacientes, divididos em 3 grupos de 50 pacientes para cada substância. Antes da higienização foi realizado um exame clínico da boca em seguida duas coletas para o isolamento de leveduras uma antes e outra após a higienização. As leveduras isoladas foram identificadas por meio do Kit API 20C AUX. Para avaliação in vitro da atividade antifúngica do digluconato de clorexidina a 0,12% e do extrato etanólico de própolis a 20% utilizou-se a técnica de microdiluição em meio RPMI 1640. O digluconato de clorexidina a 0,12% e o extrato etanólico de própolis a 6% inibiram...

Foci of infection in the mouth have been related to the impairment of the general health of the individual, arousing the interest of physicians and dentists. Infection is a frequent complication with high mortality rates in patients hospitalized in the Intensive Care Unit (ICU). These patients often do not have adequate oral hygiene, possibly due to lack of appropriate techniques for the intensive therapy teams, and the absence of relationship between dentists and nurses. Research on natural products targets the effective treatment of these infections. In dentistry it has been studied the pharmacological activity of propolis extract in some situations , such as gingivitis , periodontitis , oral ulcers, pulp mummification. Also, it has been used in pre-and post-surgical dressings and treatments of candidiasis, oral herpes and oral hygiene, due to the antiseptic and healing capacity in hospitalized individuals. The aim of this study was to evaluate oral hygiene with filtered water, chlorhexidine digluconate 0.12% and ethanol extract of propolis to 6% in ICU patients and the activity of chlorhexidine gluconate 0.12% and the ethanol extract 6% propolis envelopes yeast and subinibitory doses on the production of protease and phospholipase exoenzyme and phenotypic characteristics (fringes). 150 patients divided into 3 groups of 50 patients were studied for each substance. A clinical examination was performed before cleaning the mouth and then two samples for the isolation of yeasts was done one before and one after the cleaning. The yeasts were identified by the API 20C AUX kit. To evaluate the in vitro antifungal activity of chlorhexidine gluconate 0.12% and the ethanol extract of propolis 20% used the microdilution in RPMI 1640 The chlorhexidine gluconate 0.12% and the ethanol extract 6% propolis inhibited the yeast crecimento the third day after the...

In vitro effectiveness of Brazilian brown propolis against Enterococcus faecalis

The aim of this study was to evaluate the in vitro antimicrobial activity of Brazilian brown propolis as an intracanal medication againstEnterococcus faecalis. Thirty dentin discs prepared from intact freshly extracted bovine maxillary central incisors were infected withE. faecalis for 21 days. The specimens were distributed into six groups according to the medicament used as follows: G1- calcium hydroxide paste; G2- Carbowax 400 (control group); G3- 20% brown propolis paste; G4- 40% brown propolis paste; G5- 20% brown propolis paste + calcium hydroxide paste; and G6- 40% brown propolis paste + calcium hydroxide paste. The experimental pastes were placed into the canal lumen and left for 14 days. After each period, irrigation was performed with sterile saline to remove the medicament, and the canals were dried with sterile paper points. The dentin chips were removed from the canals with sequential sterile round burs at low speed and were immediately collected in separate test tubes containing BHI broth. The tubes were incubated at 37°C, and microbial growth was analyzed by spectrophotometry after 15 days. All the experimental medications significantly reduced the number of viable bacteria. The G4 and G5 pastes were more effective than the G1 paste, with 35.8%, 41%, and 21.3% antibacterial activity, respectively. Brazilian brown propolis shows antibacterial capacity againstE. faecalis.

Antifungal activity of propolis against Candidaspecies isolated from cases of chronic periodontitis

This research evaluated the fungistatic and fungicidal activities of red propolis alcoholic extract (RPAE) against different Candida species isolated from chronic periodontitis cases, and compared with chlorhexidine (CHX). Nineteen samples of Candida species (C. albicans [n = 12], C. tropicalis [n = 5] andC. glabrata[n = 2]) isolated from chronic periodontitis cases were analyzed. The fungistatic and fungicidal activity of both RPAE and CHX were evaluated using fluconazole and C. parapsilosis (ATCC 6258) as a control. Fungistatic activity was analyzed based on the Clinical and Laboratory Standards Institute (CLSI) reference procedure to determine the minimum inhibitory concentrations. Fungicidal activity was established according to the absence of fungal growth on Sabouraud Dextrose Agar medium. The fungistatic and fungicidal activities of RPAE were observed, respectively, at 32-64 μg/mL and 64-512 μg/mL for C.albicans, 64 μg/mL and 64-256 μg/mL for C. glabrata, and 32-64 μg/mL and 64 µg/mL for C. tropicalis. CHX fungistatic activity was observed at concentrations of 0.003-1.92 µg/mL for C. albicans, 1.92 µg/mL for C. glabrata, and 0.03-1.92 µg/mL for C. tropicalis. Fluconazole fungistatic activity ranged between 1-64 μg/mL, and fungicidal activity occurred at 8-64 μg/mL, for the threeCandida species analyzed. All the Candidaspecies were susceptible to RPAE antifungal activity, but five samples ofC.albicans, one ofC.tropicalis and one ofC.glabrata were resistant to fluconazole antifungal activity. CHX showed fungistatic activity against all the Candida species analyzed. The antifungal potential of these substances suggests that they can be applied as an alternative treatment for diseases affected by these species.

Efeitos da própolis na proliferação de fibroblastos em lesões cutâneas de ratos / Effect of propolis on fibroblast proliferation in skin lesions in rats

O presente estudo teve por objetivo verificar o efeito tópico da própolis na proliferação de fibroblastos e a disposição e volume de fibras colágenas presentes durante o processo de reparo tecidual. Foram utilizados ratos wistar, machos, divididos em dois grupos: Grupo Controle (CC) n=16 lesão tratada com creme não-iônico; Grupo Própolis (PP) n=16 lesão tratada com creme não-iônico + Própolis 10%. Nos 4º, 7°, 14° e 21° dias de tratamento foram sacrificados 4 animais de cada grupo em câmara de gás carbônico. O tecido lesionado foi coletado e fixado em formalina a 10% por 48 horas, incluído em álcool a 70%, fixado em parafina e depositado em lâminas para análise histológica. Os resultados demonstraram um aumento no número de fibroblastos e também maior e melhor disposição de fibras colágenas no grupo PP em relação ao grupo CC. Assim, as evidências obtidas no estudo mostraram que o efeito da própolis na aceleração do processo de reparo tecidual não é somente por sua ação antiinflamatória, conforme diversos estudos demonstram, mas também por sua ação direta sobre a proliferação de fibroblastos, acelerando a reversão de fibrócito para fibroblasto, e, consequentemente favorecendo a síntese e deposição de fibras colágenas, melhorando o reparo tecidual e reduzindo o tempo de cicatrização...

This study aimed to verify the effect of topical propolis on fibroblast proliferation and disposicao and volume of collagen fibers present in the tissue repair process. We used male Wistar rats were divided into two groups: control (CC) n = 16 lesions treated with non-ionic cream; Propolis Group (PP) n = 16 lesion treated with non-ionic cream + 10% propolis. At 4, 7, 14 and 21 days of treatment where sacrificed four animals from each group in a carbon dioxide chamber. The injured tissue was collected and fixed in 10% formalin for 48 hours, then in 70% alcohol, embedded in paraffin and placed on slides for histological analysis. The results showed an increase in the number of fibroblasts and also bigger and better arrangement of collagen fibers in PP group than in CC group. Thus, the evidence obtained in the study showed that the effect of propolis to speed up tissuerepair process is not only for its anti-inflammatory action, as several studies show, but also by its direct action on the proliferation of fibroblasts, accelerating the rate at which fibrocytes revert to fibroblasts, and collagen fiber arrangement, improving tissue repair and reducing the healing time...

Potential chemoprotective effects of green propolis, L-lysine and celecoxib on bone marrow cells and peripheral blood lymphocytes of Wistar rats subjected to bladder chemical carcinogenesis

PURPOSE: To evaluate the genotoxicity of propolis and L-lysine, as well as their effects on the possible cellular damage in erythroblasts (bone marrow) and leukocytes (peripheral blood) caused by the carcinogen BBN (n - butyl - n {4 - hydroxybutyl} nitrosamine) in rats subjected to bladder carcinogenesis and treated with green propolis and L-lysine. METHODS: One hundred and twenty five rats were distributed into the following groups: I, IIA, IIB, III, K, L M N, X, XI, XII and XIII. Groups I to X received BBN in drinking water for 14 weeks (wks). Group I was treated with intragastric (ig) propolis at 150 mg/kg body weight, for 44 wks, beginning 30 days before start of BBN. Groups IIA and III were treated with propolis (150 mg/kg), for 40 wks, subcutaneous (sc) and ig, respectively, beginning simultaneously with BBN. On the 32nd wk, the animals of groups L, M and N were treated ig with L-lysine (300 mg/kg), celecoxib (30 mg/kg) and propolis (300 mg/kg), respectively, up to the 40th wk. The groups that received only BBN (IIB and K) were treated with water, sc and orally, respectively, for 40 wks. Groups XI, XII and XIII received respectively propolis (150 mg/kg), L-lysine (150 mg/kg) and water ig for 40 wks. After 40 wks, the surviving animals were anesthetized and subjected to femoral bone marrow aspiration and blood collection from the aorta, for CA and MNT, respectively, for investigation of genotoxicity. RESULTS: Groups IIB and K, which received only BBN and water, showed the greatest DNA damage in peripheral leukocytes (CA) and largest number of micronuclei in bone marrow erythrocytes (MNT) in relation to all other groups that received BBN and lysine and/or propolis (p<0.001). CONCLUSIONS: Both propolis and L-lysine are effective in protecting against genotoxicity, as well not being genotoxic themselves toward the cells evaluated, at the doses and times administered and according to the two tests utilized. .