Brazilian Green Propolis Extract Research Articles

Efficacy of toothpaste containing Brazilian green propolis extracts with an optimal kaempferide/betuletol ratio for improving oral microbiota: A randomized, controlled, paired crossover study

Ethnopharmacological relevancePropolis is a resinous substance collected by honeybees from various plant sources and has been used in traditional folk medicine for centuries. Propolis has various biological properties, including antibacterial, antiviral, anti-inflammatory, and anti-tumor properties. The use of propolis in oral health care is attributable to its antimicrobial and anti-inflammatory effects. However, limited evidence exists on the in vivo efficacy of propolis against periodontal pathogens. Aim of the studyWe aimed to evaluate the efficacy of Brazilian green propolis (BGP)-containing toothpaste for improving the oral environment and define its antibacterial compounds. Materials and methodsOverall, 48 student volunteers aged 18–40 years (24 females and 24 males) were randomly categorized into the BGP and placebo groups. The BGP and placebo groups received toothpaste with and without BGP, respectively. After a baseline assessment, the plaque index (PI) score, gingival index (GI) score, and proportion of periodontal pathogens on the tongue surface were analyzed at 0, 1, and 2 weeks. Antibacterial compounds were identified using liquid-liquid partitioning, high-performance liquid chromatography purification, and nuclear magnetic resonance methods. ResultsThe concentration of BGP in the toothpaste was set at 0.0347 w/v%. Compared with the placebo group, the BGP group demonstrated a reduction in the PI score (p < 0.05) but not in the GI score, as well as a reduction in Porphyromonas gingivalis (Pg)/Total bacteria (Tb), Fusobacterium nucleatum (Fn)/Tb, and Aggregatibacter actinomycetemcomitans (Aa)/Tb (p < 0.05) but not in Streptococcus salivalius/Tb. Effect sizes for Pg, Fn and Aa were 0.360, 0.556, and 0.164, respectively. The antibacterial compounds of the BGP-containing toothpaste included a mixture of kaempferide/betuletol. ConclusionsWe confirmed the efficacy of propolis toothpaste with an optimal kaempferide/betuletol ratio for improving oral microbiota, thereby suggesting that BGP toothpaste is clinically useful in maintaining oral health and preventing periodontal disease.

Countercurrent chromatography isolation of green propolis biomarkers: Potential blockers of SARS-COV-2 RBD and ACE2 interaction

Propolis is a natural resinous mixture produced by honeybees with numerous biological activities. Considering the recently reported potential of propolis as an adjuvant in COVID-19 treatment, a methodology for the fractionation of the hexane extract of Brazilian green propolis (HEGP) was developed for the obtention of prenylated biomarkers by countercurrent chromatography. The inhibition of the interaction between the receptor binding domain (RBD) of spike and ACE2 receptor was evaluated by the Lumitᵀᴹ immunoassay. Fractionation of HEGP was performed by both normal (CCC1 and CCC2, with extended elution) and reversed (CCC3) phase elution-extrusion modes with the solvent system hexane–ethanol–water 4:3:1. The normal elution mode of CCC1 (471 mg HEGP in a 80 mL column volume, 1.6 mm id) was scaled-up (CCC5, 1211 mg HEGP in a 112 mL column volume, 2.1 mm id), leading to the isolation of 89.9 mg of artepillin C, 1; 52.7 mg of baccharin, 2; and 26.6 mg of chromene, with purities of 93 %, 83 % and 88 %, respectively, by HPLC-PDA. Among the isolated compounds, artepillin C, 1, and baccharin, 2, presented the best results in the Lumitᵀᴹ immunoassay, showing 67% and 51% inhibition, respectively, at the concentration of 10 μM. This technique proved to be of low operational cost and excellent reproducibility.

A validated ultra-performance liquid chromatography with tandem mass spectrometry method for the quantification of Brazilian green propolis main compounds

Brazilian green propolis is used in folk medicine because of its various biological properties. The hydroalcoholic extract of Brazilian green propolis is characteristic for possessing several pharmacological properties. Phytochemical investigations have attributed some of these properties to the presence of compounds, which were chosen as analytical markers. This paper reports the development and analytical validation using UPLC-MS/MS in MRM mode. Veratraldehyde was used as an internal standard in qualitative and quantitative analyses of the extracts. Relative standard deviation values obtained for intra-day and inter-day precision were lower than 4%. Of the five parameters for robustness, wavelength detection and flow rate were the critical ones. Limits of detection and quantification ranged from 0.300 to 39.500 ng.mL−1 and from 1.400 to 85.00 ng.mL−1, respectively. The recoveries were between 94.00 and 119.00%, with relative standard deviation values around 5.0%. The developed method is precise, sensitive, and reliable for analysing Brazilian green propolis.

Collapsing Glomerulopathy (CG) in a Patient with High-Risk Apolipoprotein L1 (APOL1) Genotype: 4-Year Clinical Evolution of Use of Standardized Brazilian Green Propolis Extract (EPP-AF)

Collapsing Glomerulopathy (CG) in a Patient with High-Risk Apolipoprotein L1 (APOL1) Genotype: 4-Year Clinical Evolution of Use of Standardized Brazilian Green Propolis Extract (EPP-AF)

Standardized Brazilian green propolis extract (EPP-AF®) in COVID-19 outcomes: a randomized double-blind placebo-controlled trial

SARS-CoV-2 and its different variants caused a “wave and wave” pandemic pattern. During the first wave we demonstrated that standardized Brazilian green propolis extract (EPP-AF®) reduces length of hospital stay in adult patients with COVID-19. Afterwards, we decided to evaluate the impact of EPP-AF in hospitalized patients during the third wave of the pandemic. BeeCovid2 was a randomized, double-blind, placebo-controlled clinical trial in hospitalized COVID-19 adult patients. Patients were allocated to receive an oral dose of 900 mg/day of EPP-AF® or placebo for 10 days. The primary outcome was length of hospital stay. Secondary outcomes included safety, secondary infection rate, duration of oxygen therapy dependency, acute kidney injury and need for intensive care. Patients were followed up for 28 days after admission. We enrolled 188 patients; 98 were assigned to the propolis group and 90 to the placebo group. The post-intervention length of hospital stay was of 6.5 ± 6.0 days in the propolis group versus 7.7 ± 7.1 days in the control group (95% CI − 0.74 [− 1.94 to 0.42]; p = 0.22). Propolis did not have significant impact on the need for oxygen supplementation or frequency of AKI. There was a significant difference in the incidence of secondary infection between groups, with 6.1% in the propolis group versus 18.9% in the control group (95% CI − 0.28 [0.1–0.76], p = 0.01). The use of EPP-AF was considered safe and associated with a decrease in secondary infections. The drug was not associated with a significant reduction in length of hospital stay. ClinicalTrials.gov (NCT04800224).

Effects of Three Different Brazilian Green Propolis Extract Formulations on Pro- and Anti-Inflammatory Cytokine Secretion by Macrophages

Propolis is known for its immunomodulatory properties. We investigated the effects of three recently developed propolis extract formulations: polar propolis fraction (PPF), soluble propolis dry extract (PSDE), and microencapsulated propolis extract (MPE), and some of their components, on pro- and anti-inflammatory cytokine production in a macrophage model. Bone marrow cell-derived macrophages (BMDM) in cell culture were E. coli lipopolysaccharide (500 ng/mL) stimulated for two hours and subsequently incubated for 20 hours with one of the three propolis extract formulations (1, 10, 25, 50, 100 and 300 µg/mL) or with isolated propolis components (caffeic acid, p-coumaric acid, artepillin C, or baccharin) (10, 25, 50 and 100 µg/mL) to determine how they affected secretion of the pro-inflammatory cytokines IL-6 and TNF-α, and the anti-inflammatory cytokine, IL-10. PPF increased IL-6 and IL-10 levels. PSDE increased IL-6 and IL-10 at lower concentrations, while at higher concentrations it increased TNF-α and decreased IL-10. MPE increased IL-10. Caffeic acid and PPF increased both IL-6 and IL-10. Artepillin C and PSDE decreased IL-10. Baccharin and MPE increased IL-10. Baccharin also decreased IL-6. p-coumaric acid did not affect secretion of these cytokines. Pro- and anti-inflammatory cytokine production by the different propolis extracts differed; however, all three propolis extract formulations have potential as immunomodulatory agents in food supplement and pharmaceutical products.

The effect of Brazilian Green Propolis extract on inflammation in patients with chronic kidney disease on peritoneal dialysis: A randomised double-blind controlled clinical trial

BackgroundChronic kidney disease (CKD) patients on dialysis display a low-grade systemic inflammatory burden. Nutritional interventions designed to activate the cytoprotective nuclear factor erythroid-2-related factor 2 (Nrf2) and inhibit nuclear factor-kB (NF-κB) have been proposed to mitigate this burden. Several bioactive compounds have been investigated to achieve this, including propolis, a resin produced by Apis mellifera bees. Considering the safety and efficacy of propolis, it could be a strategy to benefit these patients. Still, there are no studies using propolis in patients with CKD on peritoneal dialysis (DP), and clinical studies to support this application are lacking. Hypothesis/PurposeThe objective and novelty of the present study are to evaluate the effects of propolis supplementation on inflammatory markers in patients with CKD on PD. Study designA longitudinal, double-blind, placebo-controlled trial with CKD patients on PD. MethodsThe patients were randomised into two groups: propolis that received four capsules of 100 mg (400 mg/day), containing concentrated and standardised dry EPP-AF® Brazilian green propolis extract) or placebo, four capsules of 100 mg (400 mg/day), of magnesium stearate, silicon dioxide, and microcrystalline cellulose, for two months. Plasma levels of inflammatory cytokines, including tumour necrosis factor (TNF-α) and interleukin-6 (IL-6), were evaluated by ELISA. Quantitative real-time PCR analyses were performed to evaluate the transcriptional expression levels of Nrf2 and NF-κB in peripheral blood mononuclear cells (PBMCs). Plasma malondialdehyde (MDA) levels, a lipid peroxidation marker, was measured as thiobarbituric acid reactive substances (TBARS). Routine biochemical markers, including C-reactive protein (CRP), were analysed using commercial kits. Carotid Intima-Media Thickness (CIMT) was measured with a doppler ultrasonography device. The study was registered on ClinicalTrials.gov under the number NCT04411758. ResultsA total of 19 patients completed the study, ten patients in the propolis group (54 ± 1.0 years, five men, 7.2 (5.1) months on PD) and 9 in the placebo group (47.5 ± 15.2 years, three men, 10.8 (24.3) months on PD). The plasma levels of TNF-α reduced significantly (p = 0.02), and expression of Nrf2 showed a trend to increase (p = 0.07) after propolis supplementation. ConclusionEPP-AF® Green Propolis extract (400 mg/day) supplementation for two months appears as a potential strategy to mitigate inflammation, reducing TNF-α plasma levels in CKD patients on PD.

Novel synthetic derivatives of cinnamic and p-coumaric acids with antiproliferative effect on breast MCF-7 tumor cells

p-Coumaric acid is derived from cinnamic acid and is one of the major compounds in the Brazilian green propolis extract. Studies have shown that both p-coumaric acid and cinnamic acid have promising antiproliferative effects. In this context, aiming to increase the complexity of these active natural products and their activities, we performed coupling reactions with propargylamine and benzylamine, as well as with threonine, phenylalanine and lysine amino acids, aiming to enhance their antiproliferative effects towards the hormone-dependent breast cancer MCF-7 cells. Overall, the p-coumaric acid coupling with L-threonine amino acid (compound 15) had the best selectivity index (SI = 5.1), with half-maximal inhibitory concentration of 39.6 ± 1 μM, showing a high selectivity against hormone-dependent breast cancer cell lines MCF-7 and low cytotoxicity against the normal breast cell lines MCF-10A. Thus, this new natural product derivative may represent a prototype for the future development of antiproliferative agents, especially against hormone-dependent breast cancer.

Leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF®) and a gel formulation in a pre-clinical model.

Leishmaniasis is a widespread group of neglected vector-borne tropical diseases that possess serious therapeutic limitations. Propolis has been extensively used in traditional medical applications due to its range of biological effects, including activity against infectious agents. Here we evaluated the leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF®) and a gel formulation incorporating EPP-AF®, in both in vitro and in vivo models of Leishmania amazonensis infection. Propolis extract, obtained from a standardized blend following hydroalcoholic extraction, showed the characteristic fingerprint of Brazilian green propolis as confirmed by HPLC/DAD. A carbopol 940 gel formulation was obtained containing propolis glycolic extract at 3.6% w/w. The release profile, assessed using the Franz diffusion cell protocol, demonstrated a gradual and prolonged release of p-coumaric acid and artepillin C from the carbomer gel matrix. Quantification of p-coumaric acid and artepillin C in the gel formulation over time revealed that p-coumaric acid followed the Higuchi model, dependent on the disintegration of the pharmaceutical preparation, while artepillin C followed a zero-order profile with sustained release. In vitro analysis revealed the ability of EPP-AF® to reduce the infection index of infected macrophages (p < 0.05), while also modulating the production of inflammatory biomarkers. Decreases in nitric oxide and prostaglandin E2 levels were observed (p < 0.01), suggesting low iNOS and COX-2 activity. Furthermore, EPP-AF® treatment was found to induce heme oxygenase-1 antioxidant enzyme expression in both uninfected and L. amazonensis-infected cells, as well as inhibit IL-1β production in infected cells (p < 0.01). ERK-1/2 phosphorylation was positively correlated with TNF-α production (p < 0.05), yet no impact on parasite load was detected. In vivo analysis indicated the effectiveness of topical treatment with EPP-AF® gel alone (p < 0.05 and p < 0.01), or in combination with pentavalent antimony (p < 0.05 and p < 0.001), in the reduction of lesion size in the ears of L. amazonensis-infected BALB/c mice after seven or 3weeks of treatment, respectively. Taken together, the present results reinforce the leishmanicidal and immunomodulatory effects of Brazilian green propolis, and demonstrate promising potential for the EPP-AF® propolis gel formulation as a candidate for adjuvant therapy in the treatment of Cutaneous Leishmaniasis.

Potential Use of Brazilian Green Propolis Extracts as New Photosensitizers for Antimicrobial Photodynamic Therapy against Cariogenic Microorganisms

The synergic effect of Streptococcus mutans and Candida albicans increases dental caries severity. Antimicrobial photodynamic therapy (aPDT) is a non-invasive treatment for antimicrobial aspects. However, the current photosensitizers (PS) have many downsides for dental applications. This study aimed to evaluate the efficiency of two different Brazilian green propolis (BGP-AF and BGP-AG) as PS for aPDT against these microorganisms. A single-species biofilm was irradiated with crude extracts and their fractions and controls. Such extracts showed the best results and were evaluated in dual-species biofilms. Photodegradation, reactive oxygen species (ROS), cytotoxicity, and color stability assays were also investigated. Reductions higher than 3 log10 CFU/mL (p < 0.0001) occurred for crude BGP in single- and dual-species biofilms. Singlet oxygen was produced in BGP (p < 0.0001). BGP-mediated aPDT delayed S. mutans and C. albicans regrowth after 24 h of treatment (p < 0.0001). Both BGP did not change the color of dental materials (p > 0.05). BGP-AF-mediated aPDT showed 72.41% of oral keratinocyte viability (p < 0.0001). BGP extracts may be used in aPDT against S. mutans and C. albicans. Specifically, BGP-AF may represent a promising PS for dental applications.

Applicability of an HPLC method for analysis of alcoholic and glycolic Brazilian green-propolis extracts

Brazilian green propolis has been widely used in food and pharmaceutical products due to its valuable source of phenolic compounds and versatile biological activities. The development and validation of analytical methods are extremely useful for the characterization and quality control of products containing propolis. Therefore, the aim of this study was to optimize, validate and investigate the applicability of a reversed-phase HPLC method for analysis of different types of Brazilian green propolis extracts (glycolic and ethanolic). The method showed to be selective for the propolis phenolic markers. The analysis of variance and residues demonstrated that the method had significant linear regression, without lack of fit. It was also a precise, accurate and robust method, which was of utmost importance to analyze both glycolic and ethanolic extracts and at different concentrations. Moreover, as these products can display most complex matrices to analyze, a valid HPLC method can also prove to be specific and versatile.

Brazilian green propolis extracts modulate cholesterol homeostasis in a preclinical guinea pig model: an in vitro and in vivo study.

Green propolis produced by Apis melífera bees, having Baccharis dracunculifolia D.C. (Asteraceae) as the primary botanical source, has been used in traditional medicine to treat numerous disorders. However, studies evaluating propolis' potential in treating cardiovascular diseases via its effects on cholesterol metabolism are lacking. Therefore, this study investigated the effects of green propolis extracts on lipid metabolism in hypercholesterolemic guinea pigs. Chemical characterization of ethanolic extracts of green propolis samples was undertaken using HPLC. The in vitro characterization included an evaluation of the antioxidant capacity of the hydroalcoholic extract of green propolis (DPPH and FRAP assays) and its ability to act as an inhibitor of the HMG-CoA reductase enzyme. In vivo, we investigated the effect of the hydroalcoholic extract of green propolis on lipid metabolism in hypercholesterolemic guinea pigs. Results obtained validated previous reports of significant antioxidant activity. HPLC analysis confirmed that coumaric acid, artepillin C, and baccharin were the most common and abundant compounds in green propolis samples among the studied compounds. Furthermore, the compounds in these extracts acted as effective HMG-CoA reductase inhibitors in vitro. In vivo assays demonstrated that a hypercholesterolemic diet significantly reduced serum levels of the HDL cholesterol fraction. Simvastatin and propolis hydroalcoholic extracts promoted a significant increase in HDL cholesterol, suggesting that these extracts can improve the serum lipid profile of hypercholesterolemic guinea pigs. Results obtained in this study provide a perspective on the possible hypocholesterolemic effect of green propolis, suggesting that it can improve the serum lipid profile in hypercholesterolemic guinea pigs.

Effects of Standardized Brazilian Green Propolis Extract (EPP-AF®) on Inflammation in Haemodialysis Patients: A Clinical Trial.

Patients on haemodialysis (HD) present a significant inflammatory status, which has a pronounced negative impact on their outcomes. Propolis is a natural resin with anti-inflammatory and immunomodulatory properties. We assessed the safety and impact of a standardized Brazilian green propolis extract (EPP-AF®) on the inflammatory status in patients under conventional HD. Patients were assigned to receive 200 mg/day of EPP-AF® for 4 weeks followed by 4 weeks without the drug, and changes in plasma levels of interleukins (ILs), interferon gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α), and high-sensitivityc-reactive protein (HsCRP) were measured. A heatmap was used to illustrate trends in data variation. In total, 37 patients were included in the final analysis. Patients presented an exacerbated inflammatory state at baseline. During EPP-AF® use, there was a significant reduction in IFN-γ (p=0.005), IL-13 (p=0.04 2), IL-17 (p=0.039), IL-1ra (p=0.008), IL-8 (p=0.009), and TNF-α (p < 0.001) levels compared to baseline, and significant changes were found in Hs-CRP levels. The heatmap demonstrated a pattern of pronounced proinflammatory status at baseline, especially in patients with primary glomerulopathies, and a clear reduction in this pattern during the use of EPP-AF®. There was a tendency to maintain this reduction after suspension of EPP-AF®. No significant side effects were observed. Patients under haemodialysis presented a pronounced inflammatory status, and EPP-AF® was demonstrated to be safe and associated with a significant and maintained reduction in proinflammatory cytokines in this population. This trial is registered with Clinicaltrials.gov NCT04072341.

The Effects of Brazilian Green Propolis Extract on Inflammation in CKD on Peritoneal Dialysis (PD): A Randomized Double-Blind Controlled Clinical Trial

The Effects of Brazilian Green Propolis Extract on Inflammation in CKD on Peritoneal Dialysis (PD): A Randomized Double-Blind Controlled Clinical Trial

Purification of Artepillin C (ARC) with Anticancer Activities from Supercritical CO2-Ethanol Extracts of Brazilian Green Propolis by Using Adsorption Column Chromatography

Phenolic compounds with varying phenolic structures are present in both terrestrial and marine organisms. Artepillin C (ARC) is a prenylated phenolic compound produced specifically in Brazilian green propolis. Earlier studies have demonstrated the potential of ARC as a dietary supplement for cancer prevention due to its ability to suppress tumor growth and metastasis. Here we report the development of an effective adsorption chromatography combined with supercritical CO2 extraction for the massive purification of ARC. A mixture of supercritical CO2 and ethanol extract of Brazilian green propolis was passed through a stainless chromatographic column packed with protruding metals at 4,000 psi and 50 °C to remove wax from the column’s base. ARC mixed with flavonoids collected from the top of the column was applied to another stainless column filled with silica gel under similar working conditions. After flavonoids were removed, strongly adsorbed ARC at the top portion of the column was collected by ethanol elution. The purity of ARC isolated using this method was between 90% and 95%, as indicated by HPLC chromatograms. Purified ARC strongly inhibited cancer cell division based on its cytotoxic effects on HCT116 human colon cancer cells, resulting in a significantly increased distribution of cells in the S phase. ARC also blocks the motility of HCT116 cells, indicating its ability to suppress tumor metastasis. The developed chromatographic purification protocol can be used to isolate prenylated phenolic compounds from marine organisms.

Brazilian Green Propolis Modulates Cholesterol Homeostasis in a Preclinical Guinea Pig Model

This study investigates the effect of the hydroalcoholic extract of Brazilian green propolis on lipid metabolism in hypercholesterolemic guinea pigs. This product has been considered an important source of bioactive compounds with potential health benefits. Thirty‐six guinea pigs, Cavia porcellus (male), 90 days old, weight of 400 g, were divided into six groups of six animals each and fed a standard AIN‐93 M diet (C), hypercholesterolemic diet (H), hypercholesterolemic diet + hydroalcoholic extract (75 mg/kg body weight [HEtOH1]; 150 mg/kg body weight [HEtOH2], and 300 mg/kg body weight [HEtOH3]), or hypercholesterolemic diet + simvastatin (1.5 mg/kg body weight) (HS) for 10 weeks. The group‐specific diet and water were provided ad libitum throughout the experimental period. Blood and liver tissue samples were collected for biochemical analysis and histopathology. Statistical analysis was performed using GraphPad Prism software version 5.0. The experimental protocol for the use of animals was approved by the Ethics Committee on Animal Use of the University of São Paulo (N°18.1.999.60.0). Animals in the H groups had a lower food intake than in the C group. Fecal excretion was regulated by the administration of extracts, being higher in these groups when compared to H and HS groups. Liver weights were influenced by the diet and administration of the hydroalcoholic extract of green propolis. Hypercholesterolemic diets determined an increase of about 2 times in the relative weight of this organ when compared to the C group, being more expressive in the groups of animals that received the hydroalcoholic extract. A significant increase in serum levels of total and non‐HDL cholesterol was observed in H group, concomitant with a reduction in the concentration of cholesterol‐HDL when compared to group C. However, these high levels of total cholesterol were reduced when simvastatin and extracts were administered with the hypercholesterolemic diet. The same was observed for the non‐HDL cholesterol fraction, where administration with simvastatin and extracts significantly reduced the concentration of these fractions, especially in HS and HEtOH3 groups when compared to group H. Analysis of the HDL fraction demonstrates that the animals submitted to the hypercholesterolemic diet had the lowest levels for this parameter when compared to the groups that received the control diet and/or extracts and simvastatin. There was a significant reduction in serum triacylglycerol levels in the HEtOH2 and HEtOH3 groups. However, a change in liver metabolism was observed, favoring the deposition of lipids in the liver, a different profile from what was observed when administering simvastatin. C group presented a histological aspect compatible with the normality of the organ. The H and HS groups presented moderate to severe fat degeneration, with a slight degeneration in the HS group when compared to the H group. A profile similar to that of the HS group was observed in the HEtOH2 group. Our results suggest that the Brazilian green propolis extract induces positive effects on dyslipidemia.

Brazilian Green Propolis‐Based Nanostructured System: Development, Characterization and its Role in the Modulation of Cytokines in In Vitro and In Vivo Experimental Models

Brazilian green propolis from Baccharis dracunculifolia (“alecrim do campo”), stands out for the majority presence of phenolic acids with therapeutic properties well‐described in the literature, such as anti‐inflammatory, antioxidant, and antitumor. However, its direct application, like many natural products, is limited due to its unfavorable physicochemical properties. Thus, we hypothesize that the encapsulation of these compounds in nanostructured systems is a suitable instrument to overcome these limitations. In this work, an oil/water (O/W) nanoemulsion containing Brazilian green propolis extract (NE‐BGP) was developed, and its bioactivity was evaluated in vivo and in vitro experimental models. Quantitative analysis by HPLC showed the presence of coumaric acid, artepelin C and baccharin in the standardized extract obtained, corresponding to 3.2%, 16.8% and 2.3% (w/w), respectively. The developed nanoemulsion showed high stability evaluated for 90 days, with the size around 130 nm, PdI less than 0.3, and negatively charged zeta potential (‐20 mV). The encapsulation efficiency of the markers evaluated in NE‐BGP determined by the validated UPLC‐MS method was superior to 97% for 90 days. Morphological analysis using transmission electron microscopy (TEM) showed globular and spherical shapes. The cytotoxicity was evaluated in 3T3 (murine fibroblasts) and AMJ2‐C11 (murine alveolar macrophages) cells and showed IC50 between 20.88 and 49.32 µg/mL after 24 hours of treatment by the neutral red method, respectively. The in vitro anti‐inflammatory activity in AMJ2‐C11 cells stimulated with LPS 5 µg/mL showed a significant reduction in the levels of nitric oxide (NO), as well as cytokines quantified by CBA (Cytometric Bead Array), IFN‐γ, IL‐4, and IL‐6 in cells treated with NE‐BGP (15 µg/ml) when compared to the control group. Male Balb/C mice (n=5) were used to evaluate the in vivo anti‐inflammatory activity. NE‐BGP administered orally at 100mg/Kg showed a reduction in edema, in the number of inflammatory cells in the joints, and an increase in the mechanical threshold reducing nociceptive stimuli in zymosan‐induced mice arthritis. These results indicate that the developed formulation is an exciting system to encapsulate green propolis extract with high physicochemical stability, anti‐inflammatory and analgesic effects, suggesting NE‐BGP is a possible candidate for the arthritis treatment.

The use of standardized Brazilian green propolis extract (EPP-AF) as an adjunct treatment for hospitalized COVID-19 patients (BeeCovid2): a structured summary of a study protocol for a randomized controlled trial

BackgroundThe 2019 coronavirus disease (COVID-19) pandemic continues to spread and affects large numbers of people with unprecedented impacts. Experimental evidence has already been obtained for use of the standardized extract of Brazilian green propolis (EPP-AF) against viral targets, and clinical rationality has been demonstrated for testing this extract as an adjunct to treatment in patients affected by COVID-19. The BeeCovid2 study aims to assess whether EPP-AF has an impact on the improvement of patients hospitalized with COVID-19 by reducing the length of hospital stay.MethodsBeeCovid2 is a randomized, double-blinded, placebo-controlled clinical study being conducted in Brazil to provide further evidence on the effectiveness of standardized green propolis extract as an adjunctive treatment for adults hospitalized with COVID-19. Hospitalized patients over 18 years of age with a confirmed diagnosis of COVID-19 and up to 14 days of symptoms were included. Patients under mechanical ventilation at randomization, pregnant women, cancer patients, transplanted or using immunosuppression, HIV patients, patients who used propolis in the last 30 days, bacterial or fungal infection at randomization, impossibility of using medication orally or enterally, and advanced chronic diseases (e.g., advanced heart failure, severe liver disease, and end-stage chronic kidney disease). Enrolled patients are randomized at a 1:1 ratio to receive placebo or standardized propolis extract (900 mg/day) for 10 days. The study treatments are administered in a double-blinded manner, and patients are followed for 28 days. The primary outcome is the difference in length of hospital stay in days between groups. Secondary outcomes include the need for mechanical ventilation, the rate of secondary infection, rate of acute kidney injury, the need for renal replacement therapy, the requirement for vasoactive drugs, the use of an intra-aortic balloon pump (IABP), and the use of extracorporeal membrane oxygenation (ECMO).DiscussionThis trial is very useful and will provide more data on the effectiveness of using the standardized Brazilian green propolis extract as an adjunctive treatment in association with standard care in adults hospitalized with moderate to severe acute COVID-19.Trial registrationClinicalTrials.gov NCT04800224. Registered on March 16, 2021.

Brazilian green propolis reduces worm burden and hepatic granuloma formation in a Schistosoma mansoni experimental murine model.

Characterized as an acute and chronic parasitic disease, schistosomiasis mansoni has as its central pathology the formation of hepatic granulomas in response to the parasite's eggs trapped in the host's liver. In recent years, research on propolis has grown; however, there is little anthelmintic work on this bee product. In the propolis scenario, Brazilian ones receive attention, with green and red propolis standing out. This study aims to evaluate in vivo the standardized extract of Brazilian green propolis (Pex) against Schistosoma mansoni. The in vivo antiparasitic activity of Pex was conducted in female BALB/c mice infected with S. mansoni and of the three groups treated with Pex (300mg/kg); G2 (35th to 42nd dpi) reduced the total worm burden by 55.32%, followed by G3 (42nd to 49th dpi) and G4 (49th to 56th dpi), with about 46%. Furthermore, G2 significantly reduced the total egg load in the ileum (59.33%) and showed an increase in the dead eggs. Similarly, histological analysis of the livers showed a significant reduction in the number and diameter of the granulomas. Based on these results, there is an interesting schistosomicidal activity of Pex and its potential against the formation of hepatic granulomas, paving the way for more detailed studies of propolis in the animal model of schistosomiasis mansoni.

Antioxidant activity, sensory analysis and acceptability of red fruit juice supplemented with Brazilian green propolis

Abstract In this work were determining the total phenolic content, total flavonoids and antioxidant capacity in formulations of red fruit juice supplemented with different concentrations of aqueous extract of Brazilian green propolis. And, also was investigate the formulation sensory acceptance by volunteers’ consumers. The addition of propolis to red fruit juice increased the content of total phenolic compounds and flavonoids, enhanced the antioxidant activity of the final product as assessed by radical scavenging DPPH and ABTS and FRAP methods, and no negative effect on consumer taste. All red fruit juice formulations containing propolis presented significant results through overall acceptability test and were approved in relation to purchase intent by the volunteer consumers. Formulation F3 (6.1 mg.mL-1) showed the best results. The antioxidant activity increment was proportional to the concentration of aqueous green propolis extracts added to the juice. The positive purchase intention reached up 64.75% of participants' acceptance. A new, sugar-free drink with pleasurable sensory and functional properties, combined with health benefits, has been obtained.