Evaluation of Effect of Biologically Synthesized Ethanolic Extract of Propolis-Loaded Poly(-Lactic-co-Glycolic Acid) Nanoparticles on Wound Healing in Diabetic Rats

Wound healing is interaction of a complex cascade of cellular/biochemical actions leading to restoration of structural and functional integrity with regain of injured tissues strength. This study was aimed at evaluation of application of ethanolic extract of propolis-loaded poly(-lactic-co-glycolic acid) nanoparticles (EEP-PLGA NPs) on wound healing in diabetic rats. Sixty rats were randomized into four groups of 15 rats each: In control group (Control) diabetic wound was treated with normal saline. In Carrier 1 group diabetic wound was treated with PLGA nanoparticles based solution. In Carrier 2 group the diabetic wound was treated with EEP. In Treatment group animals received EEP-PLGA NPs on the wound. Wound size was measured on 7, 14 and 21 days after surgery. The expression of p53, bcl-2, Caspase III, were evaluated using reverse-transcription PCR and Immunohistochemical staining. The Treatment group had significantly reduced the wound size compared to other groups ( P = 0.001). histological and morphometric studies, and mean rank of the qualitative studies demonstrated that there was significant difference between Treatment group and other groups ( P < .05). Observations demonstrated that ethanolic extract of propolis-loaded PLGA nanoparticles significantly shortened the inflammatory phase and accelerated the cellular proliferation. Accordingly, the animals in Treatment group revealed significantly ( P < .05) higher fibroblast distribution/one mm2 of wound area and rapid re epithelialization. The mRNA levels of bcl-2, p53 and caspase III were remarkably ( P < .05) higher in Treatment group compared to control and animals. The immunohistochemical analyzes confirmed the RT-PCR findings. EEP-PLGA NPs offered potential advantages in wound healing acceleration and improvement through angiogenesis stimulation, fibroblast proliferation and granulation tissue formation in early days of healing phases, acceleration in diabetic wound repair associated with earlier wound contraction and stability of damaged area by rearrangement of granulation tissue and collagen fibers.

THE COMPARISON OF PROPOLIS EFFECTS ON TUMOR NECROSIS FACTOR ALPHA AND MALONDIALDEHYDE BETWEEN INHALATION AND CUTANEOUS ANTHRAX ANIMAL MODELS

Background: Inflammatory response and oxidative stress can be found in anthrax characterized by increased level of serum Tumor Necrosis Factor Alpha (TNF-α) and Malondialdehyde (MDA). The use of antibiotics in anthrax has been known to cause some disturbing side-effects, such as allergic reaction, nausea, vomiting, and antibiotic resistance. Thus, ethanolic extract of propolis (EEP) might be the alternative regimen, due to its anti-inflammatory and antioxidant properties. This study aimed to compare the effects of ethanolic extract of propolis (EEP) on TNF-α and MDA between the inhalation and cutaneous anthrax animal model. Materials and Methods: This was an experimental study with a post-test-only control group design on 40 samples of Rattus norvegicus. Samples were randomized into 5 groups: control, inhalation anthrax model, inhalation anthrax model + EEP, cutaneous anthrax model, and cutaneous anthrax model + EEP. After 14 days, the level of TNF-α and MDA were measured. To compare the data, we used the ANOVA test continued by the post-hoc Turkey test. Results: The results obtained showed that the level of TNF-α and MDA between the inhalation and cutaneous anthrax animal models treated with EEP were statistically different (p < 0.05). The P5 group showed the lowest level of TNF-α (6.822 ± 0.383 pg/ml) and MDA (2.717 ± 0.383 nmol/ml). Conclusion: EEP has a better effect on reducing TNF-α and MDA in cutaneous anthrax animal models compared to the inhalation anthrax animal model.

Antimicrobial activity of a novel Spanish propolis against planktonic and sessile oral Streptococcus spp

Increased bacterial resistance to traditional antimicrobial agents has prompted the use of natural products with antimicrobial properties such as propolis, extensively employed since ancient times. However, the chemical composition of propolis extracts is extremely complex and has been shown to vary depending on the region and season of collection, due to variations in the flora from which the pharmacological substances are obtained, being therefore essential for their antimicrobial activity to be checked before use. For this purpose, we evaluate the in vitro antimicrobial and anti-biofilm activity of a new and promising Spanish ethanolic extract of propolis (SEEP) on Streptococcus mutans and Streptococcus sanguinis, responsible, as dominant ‘pioneer’ species, for dental plaque. Results reveal that S. sanguinis is more sensitive to SEEP, slowing and retarding its growth considerably with lower concentrations than those needed to produce the same effect in S. mutans. SEEP presents concentration- and time-dependent killing activity and, furthermore, some of the subinhibitory concentrations employed increased biofilm formation even when bacterial growth decreased. Mono and dual-species biofilms were also inhibited by SEEP. Findings obtained clearly show the relevance of using biofilm and subinhibitory concentration models to determine optimal treatment concentrations.

Evaluation of Effectiveness of a Toothpaste Containing Tea Tree Oil and Ethanolic Extract of Propolis on the Improvement of Oral Health in Patients Using Removable Partial Dentures

The aim of this study was to evaluate the effect of toothpaste containing natural tea tree essential oil (TTO) and ethanolic extract of propolis (EEP), on microflora and selected indicators of oral health in patients using removable acrylic partial dentures. Fifty patients with varying conditions of hygiene were divided into two groups. The study group received the toothpaste with TTO and EEP, while the control group received the same toothpaste but without TTO and EEP. At the first visit, oral hygiene and hygiene of the prostheses were carried out. Control visits took place 7 and 28 days later and compared to baseline. Indexes like API (Approximal Plaque Index), mSBI (modified Sulcus Bleeding Index), OHI-s (simplified Oral Hygiene Index), and DPI (Denture Plaque Index) were assessed in three subsequent stages, and swabs were collected from floor of the mouth area to assess the microbiota. After 7 and 28 days of using the toothpaste with TTO and EEP, a statistically significant decrease of the examined indicator values were observed in the study group as compared to the values upon the initial visit. The number of isolated strains of microorganisms in the study group was decreased or maintained at the same level, whereas in the control group an increase in the number of isolated strains was observed. The observed stabilization of oral microbiota in patients from the study group confirms the beneficial activity of toothpaste containing EEP and TTO compared to the control group.

Antifungal and anti-biofilm activity of a new Spanish extract of propolis against Candida glabrata

Background Resistance to traditional antifungal agents is a considerable health problem nowadays, aggravated by infectious processes related to biofilm formation, usually on implantable devices. Therefore, it is necessary to identify new antimicrobial molecules, such as natural products, to develop new therapeutic strategies to prevent and eradicate these infections. One promising product is propolis, a natural resin produced by honeybees with substances from various botanical sources, beeswax and salivary enzymes. The aim of this work was to study the effect of a new Spanish ethanolic extract of propolis (SEEP) on growth, cell surface hydrophobicity, adherence and biofilm formation of Candida glabrata, a yeast capable of achieving high levels of resistance to available anti-fungal agents. Methods The antifungal activity of SEEP was evaluated in the planktonic cells of 12 clinical isolates of C. glabrata. The minimum inhibitory concentration (MIC) of propolis was determined by quantifying visible growth inhibition by serial plate dilutions. The minimum fungicide concentration (MFC) was evaluated as the lowest concentration of propolis that produced a 95% decrease in cfu/mL, and is presented as MFC50 and MFC90, which corresponds to the minimum concentrations at which 50 and 90% of the C. glabrata isolates were inhibited, respectively. Influence on cell surface hydrophobicity (CSH) was determined by the method of microbial adhesion to hydrocarbons (MATH). The propolis effect on adhesion and biofilm formation was determined in microtiter plates by measurement of optical density (OD) and metabolic activity (XTT-assay) in the presence of sub-MIC concentrations of SEEP. Results SEEP had antifungal capacity against C. glabrata isolates, with a MIC50 of 0.2% (v/v) and an MFC50 of 0.4%, even in azole-resistant strains. SEEP did not have a clear effect on surface hydrophobicity and adhesion, but an inhibitory effect on biofilm formation was observed at subinhibitory concentrations (0.1 and 0.05%) with a significant decrease in biofilm metabolism. Conclusions The novel Spanish ethanolic extract of propolis shows antifungal activity against C. glabrata, and decreases biofilm formation. These results suggest its possible use in the control of fungal infections associated with biofilms.

Ethanolic Extract of Algerian Propolis Induced Cell Damage in Staphylococcus Aureus: A Promising Alternative as a Natural Bio-Preservative in Food Products

This study describes the antistaphylococcal mechanism of the ethanolic extract of Algerian propolis on Staphylococcus aureus ATCC 25923. To investigate the underlying mechanism of action of the ethanolic extract of propolis, bacteriolysis, bacterial death, leakage of potassium, proteins, nucleic components, and scanning electron microscopic studies were conducted. The results showed that the minimum inhibitory concentration (MIC) of ethanolic extract of propolis against Staphylococcus aureus ATCC 25923 was 39 μg ml–1. The extract displayed significant bactericid activity against S. aureus in a time and concentration dependant manner. Its mode of action was evident from the increase of K+ efflux and nucleotide leakage. These results were confirmed by Scanning Electron Microscopy (SEM) that showed remarkable morphological and ultrastructural changes in S. aureus after exposure to 1MIC and 2MIC concentrations. The overall study contributed to the understanding of the antistaphylococcal mechanism of ethanolic extract of propolis. It emphasizes its potential to be used as an important natural bio-preservatives in food products.

Synergistic action of propolis with levodopa in the management of Parkinsonism in Drosophila melanogaster

BackgroundThe Phosphatase and tensin-induced putative kinase 1 (PINK1B9) mutant for Drosophila melanogaster is a key tool that has been used in assessing the pathology of Parkinsonism and its possible remedy. This research was targeted toward determining the effects of ethanolic extract of propolis, with levodopa therapy in the management of Parkinsonism.MethodThe PINK1B9 flies were divided into groups and fed with the different treatment doses of ethanoic extract of propolis. The treatment groups were subjected to 21 days of administration of propolis and the levodopa at different doses after which percentage climbing index, antioxidant activity and lifespan studies were done.ResultsPropolis alone improved motor activity, antioxidant and lifespan in Drosophila melanogaster than in PINK1 flies. Propolis in combination with levodopa significantly (P<0.05) improved physiological parameters at higher than lower concentrations in Parkinsonism Drosophila melanogaster demonstrating its importance in managing side effects associated with levodopa.ConclusionPropolis is a novel candidate as an alternative and integrative medicinal option to use in the management of Parkinsonism in both animals and humans at higher concentrations.

Chemical Profile and Antibacterial Activity of a Novel Spanish Propolis with New Polyphenols also Found in Olive Oil and High Amounts of Flavonoids

Propolis is a natural product obtained from hives. Its chemical composition varies depending on the flora of its surroundings, but nevertheless, common for all types of propolis, they all exhibit remarkable biological activities. The aim of this study was to investigate the chemical composition and antimicrobial activity of a novel Spanish Ethanolic Extract of Propolis (SEEP). It was found that this new SEEP contains high amounts of polyphenols (205 ± 34 mg GAE/g), with unusually more than half of this of the flavonoid class (127 ± 19 mg QE/g). Moreover, a detailed analysis of its chemical composition revealed the presence of olive oil compounds (Vanillic acid, 1-Acetoxypinoresinol, p-HPEA-EA and 3,4-DHPEA-EDA) never detected before in propolis samples. Additionally, relatively high amounts of ferulic acid and quercetin were distinguished, both known for their important therapeutic benefits. Regarding the antimicrobial properties of SEEP, the minimal inhibitory and bactericidal concentrations (MIC and MBC) against Staphylococcus epidermidis strains were found at the concentrations of 240 and 480 µg/mL, respectively. Importantly, subinhibitory concentrations were also found to significantly decrease bacterial growth. Therefore, the results presented here uncover a new type of propolis rich in flavonoids with promising potential uses in different areas of human health.

The Impact of Water and Ethanolic Extracts of Propolis on Fatty Acid Changes of Sardine Fillets

The polyunsaturated fatty acids (PUFA) of lipids in muscle of sardine were known as unstable to oxidation. Propolis is a natural compounds produced by honey bees from substances collected from parts of plants, buds, and exudates. Nowadays, the particular attention has been attributed to propolis underlying their antioxidant properties due to the presence of the flavonoids, phenolic acids, and ethers. The impact of water and ethanolic extract of propolis at doses of 0.4 and 0.8% on fatty acid changes of vacuum packaged sardine fillets were investigated during chill storage. The main fatty acids in sardine fillets were palmitic acid (C16:0), palmitoleic acid (C16:1), myristic acid (C14:0), eicosapentaenoic acid (EPA, C20:5n3) and docosahexaenoic acid (DHA, C22:6 n3), respectively. There was no significant differences in EPA and DHA between control and treated groups during storage apart from 4 and 11th day. Initial n6/n3 ratio in sardine fillet was 0.17. Group treated with 0.4 propolis ethanolic extract comprised lower n6/n3 ratio at 4, 11 and 13th days than that of other groups. Although the effects of propolis extracts on fatty acid composition of fish were variable, application of propolis ethanolic and water extracts on fish fillets at doses of 0.8% had a positive impact due to lead a better oxidative stability of PUFA content.