Propolis is multicomponent substance collected by honeybees from various plants. It is known for numerous biological effects and is commonly used as ethanolic extract because most of active substances of propolis are ethanol-soluble. However, water-based propolis extracts could be applied more safely, as this solvent is more biocompatible. On the other hand, water extracts has significantly smaller range and quantity of active compounds. The extraction power of water could be enhanced by adding co-solvent which increases both solubility and penetration of propolis compounds. However, variation of solvents results in different composition of active substances that might have distinct effects. The majority of biological effects of propolis are attributed to the antioxidant properties of its active compounds. Antioxidant effect might be a result of either direct scavenging of ROS or modulation of ROS producing organelle activity. Therefore, the aim of this study was to investigate and compare chemical composition, antioxidant properties and effects on mitochondrial respiration of aqueous (AqEP), polyethylene glycol-aqueous (Pg-AqEP) and ethanolic (EEP) propolis extracts. Chemical composition of propolis extracts was determined using HPLC and Folin-Ciocalteu method. Ability to neutralize H2O2 and intracellular ROS concentration in C6 glioma cells were determined fluorometrically by using 10-acetyl-3,7-dihydroxyphenoxazine and 2′,7′-dichlorofluorescein diacetate, respectively. Mitochondrial superoxide generation was assessed under fluorescent microscope by using MitoSOX Red. Oxygen uptake rates of mitochondria were recorded by high-resolution respirometer Oxygraph-2 k. Our data revealed that phenolic acids and aldehydes make up 40–42% of all extracted and identified compounds in AqEP and Pg-AqEP and only 16% in EEP. All preparations revealed similar antioxidant activity in cell culture medium but Pg-AqEP and EEP demonstrated better mitochondrial superoxide and total intracellular ROS decreasing properties. At higher concentrations, AqEP and EEP inhibited mitochondrial respiration, but Pg-AqEP had concentration-dependent mitochondria-uncoupling effect. Aqueous and non-aqueous propolis extracts differ by composition, but all of them possess antioxidant properties and neutralize H2O2 in solution at similar efficiency. However, both Pg-AqEP and EEP were more effective in decreasing intracellular and intramitochondrial ROS compared to AqEP. At higher concentrations, these preparations affect mitochondrial functions and change energy production in C6 cells.

Antioxidant and free radical scavenging effects are attributed to phenolic compounds present in propolis, and when delivered to the skin surface and following penetration into epidermis and dermis, they can contribute to skin protection from damaging action of free radicals that are formed under UV and premature skin aging. This study was designed to determine the penetration of phenolic acids and vanillin into the human skinin vitrofrom experimentally designed vehicles. Results of the study demonstrated the ability of propolis phenolic acids (vanillic, coumaric, caffeic, and ferulic acids) and vanillin to penetrate into skin epidermis and dermis. The rate of penetration and distribution is affected both by physicochemical characteristics of active substances and physical structure and chemical composition of semisolid vehicle. Vanillin and vanillic acid demonstrated relatively high penetration through epidermis into dermis where these compounds were concentrated, coumaric and ferulic acids were uniformly distributed between epidermis and dermis, and caffeic acid slowly penetrated into epidermis and was not determined in dermis. Further studies are deemed relevant for the development of semisolid topically applied systems designed for efficient delivery of propolis antioxidants into the skin.

An electrospun hydrophilic non-water-soluble biocompatible polylactic acid (PLA) nonwoven material was used as a delivery system for propolis ethanolic extract (PEE) and silver nanoparticles (AgNPs) that are known for their established antiseptic and antimicrobial activity. Combination of PEE and AgNPs in a single product should provide efficient antimicrobial protection and improved wound healing. Evaluations of PEE and AgNPs on morphology of electrospun materials, release kinetics of AgNPs and phenolic compounds, antibacterial properties, and cytotoxicity of electrospun PLA materials were performed. The presence of PEE or/and AgNPs resulted in denser mats formed by thicker PLA fibers. The average diameter of PLA microfibers was 168±29 nm. The average diameter of microfibers increased to 318±40 and 370±30 nm when 10 wt% and 20 wt% ethanol were added, respectively. Addition of 10 wt% or 20 wt% PEE increased the diameter to 282±25 and 371±25 nm, respectively. Suspension of AgNPs also caused the formation of thicker microfibers with 254±25 nm diameter. Electrospun PLA microfibers with PEE maintained viability of HaCaT cells. Testing of antimicrobial activity confirmed the ability of AgNPs containing PLA electrospun materials to inhibit the growth of microorganisms.

Water-based propolis extracts usually contain up to 10-fold lower quantities of active ingredients due to poor solubility in water of propolis bioactive compounds when compared with ethanol-based extracts. Since ethanol-based extracts are of limited use, water-based extracts are preferred nowadays. The application of alternative extraction techniques should be evaluated to improve extraction efficiency. Aqueous propolis extracts were prepared using purified water and propylene glycol, 2-hydroxypropyl-beta-cyclodextrin and sodium bicarbonate aqueous solutions. A microwave-assisted extraction method was applied in cycles. The total concentration of hydroxycinnamic acids in aqueous propolis extract produced by four extraction cycles was determined to be 1502.1 ± 130.1 μg/mL and 20% propylene glycol, 10% 2-hydroxypropyl-beta-cyclodextrin and 5% sodium bicarbonate aqueous solutions, increasing the total concentration of hydroxycinnamic acids by 1.6, 1.7 and 1.9-fold, respectively. An application of microwave-assisted extraction method and the procedure of repeating extraction cycles reliably increased the quantity of hydroxycinnamic acids in aqueous propolis extracts. Similarly, the presence of propylene glycol, 2-hydroxypropyl-β-cyclodextrin and sodium bicarbonate increased the concentration of the hydroxycinnamic acids in propolis extracts.

Emulsions are thermodynamically unstable systems and it is difficult to produce biphasic formulations with large amounts of oil. The aim of our study was to prepare biphasic formulations containing 1% ciclopirox olamine and to determine the influence of the method of oil incorporation (without and with emulsifier and gelifier) on the physical (pH, particle size, rheological properties), mechanical, and biopharmaceutical properties of the formulations. It was found that the use of a poloxamer 407 gel as the hydrophase could result in a stable formulation when an oil with (EPG) or without an emulsifier (APG) or oleogel (OPG) was used as the oily phase. The results of the studies showed that the addition of an emulsifier (polysorbate 80) led to a decrease in the sol-gel temperature, a slower release of ciclopirox olamine, and a higher stability in the freeze–thaw test. However, regardless of the way the oil is incorporated, the particles are distributed in the same range and the antifungal activity against T. rubrum is the same. It is possible to create a biphasic formulation with a large amount of oil and poloxamer gel, but for greater stability, it is recommended to include an emulsifier in the composition.

Microemulsions (MEs) were formulated using PEG‐8 caprylic/capric glycerides and ethanolic propolis extracts. Characterization of MEs was performed by determining mean droplet size, polydispersity index, stability under varying external factors, and formulation effect on delivery of phenolic compounds into the skin ex vivo. Essential oils were included into the formulations of MEs and their influence on physical characteristics of the nanostructured systems as well as penetration into epidermis and dermis were evaluated. The droplet size, their distribution, and stability of the formulated MEs were not affected. Presence of essential oils in the formulation increased penetration of phenolic compounds in general, but only the amount of ferulic acid increased significantly. Mean droplet size increased with increase of oily phase amount, suggesting that phenolic compounds and components of essential oils were not modifying the formation of the interphase film composition and/or structure. Phenolic compounds were predominantly located in the lipid phase of the MEs thus minimizing their availability at the surface of the skin.