Therapeutic bullfrog oil-based nanoemulsion for oral application: Development, characterization and stability

The aim of this study was to develop, optimize, and characterize a stable therapeutic bullfrog oil based nanoemulsion for oral application using a rational experimental design approach. The optimized oral nanoemulsion contained 0.2 % sodium benzoate and 0.02 % propyl-paraben as preservatives; 0.1 % sucralose and 0.4 % acesulfam K as sweeteners and 0.1 % tutti-frutti as flavoring to mask the unpleasant organoleptic characteristics of bullfrog oil. The oral O/W-nanoemulsion showed the droplet size, PDI, zeta potential, and pH of 410 ± 8 nm, 0.20 ± 0.02, –38 ± 2.5 mV, and 6.43 ± 0.05, respectively. The optimized oral nanoemulsion showed a milky single-phase and optimal physical stability at 25 °C for 90 days. Indeed, higher oxidation induction time and lower formation of peroxides in the oral nanoemulsion were responsible for improving its stability. A therapeutic delivery system containing bullfrog oil for oral application was successfully developed and optimized with ideal thermo-oxidative stability.

Buccal Bullfrog (Rana catesbeiana Shaw) Oil Emulsion: A Mucoadhesive System Intended for Treatment of Oral Candidiasis

Oral candidiasis (OC) is an infectious disease caused by microorganisms of the genus Candida, leading to lesions in the buccal cavity. Its treatment consists of the administration of topical or systemic antifungal agents, which may compromise the patient compliance due to its side effects, highlighting the need for alternative treatments. In this scenario, bullfrog oil, an animal oil composed of a pool of saturated and unsaturated fatty acids, is introduced as a potential antifungal raw material. Thus, the aim of this work was to produce a mucoadhesive emulsified system able to deliver the bullfrog oil in the buccal cavity to treat the OC. The emulsion was produced and characterized by visual inspection, droplet size, polydispersity index (PdI), and zeta potential over the course of 60 days. In addition, its mucoadhesive ability was evaluated using an in vitro mucin model. The antifungal activity, evaluated by the broth microdilution assay and the biocompatibility, performed against human erythrocytes, were also carried out. The emulsion showed a droplet size of 320.79 ± 35.60 nm, a PdI of 0.49 ± 0.08, and a zeta potential of −38.53 ± 6.23 mV, with no significant changes over 60 days. The mucoadhesive properties of the system was improved by the use of pharmaceutical excipients. The antifungal activity showed that the bullfrog oil and the emulsion were able to inhibit the growth of different Candida species. Furthermore, the emulsion showed no significant hemolytic effect. Overall, the system showed suitable physicochemical characteristics and biocompatibility, with substantial in vitro antifungal activity, suggesting that this system can be further investigated for OC treatment.

Analysis of the Anti-Inflammatory Potential of Pure and Microemulsified Bullfrog Oil in Acute Lung Injury

Infectious diseases account for more than a third of all hospital admissions, and are highly prevalent in intensive care units. Currently, sepsis is one of the diseases with the highest morbidity and mortality rates worldwide, with death rates reaching up to 60% among intensive care patients, according to statistics from low-income countries. The prominence of multi-resistant microorganisms is rising, while the possibilities of development of new target drugs are being exhausted. Thus, the objective of the present study was to evaluate the anti-inflammatory potential of bullfrog oil in its pure state and in a microemulsion system in an experimental model of sepsis. Mice were separated into three groups and treated with bullfrog oil in its pure state, in a microemulsion, and with saline solution, and subsequently submitted to induction of sepsis. Bronchoalveolar lavages were performed for cell counts, as well as analyses of lung tissue samples. When the washings were analyzed, no statistically significant difference was observed in cell migration between the experimental groups, but a difference was observed between these groups and the saline solution group. When the lung tissue samples were analyzed, intense tissue wear was observed in the bullfrog oil groups, with the presence of cellular infiltrate and rupture of respiratory bronchioles and alveoli. However, in the microemulsion group, no major tissue wear was observed, and the pulmonary parenchyma was more preserved. Thus, we concluded that bullfrog oil in pure form and in a microemulsion system are good modulators of the inflammatory response, with the microemulsion system being more efficient in protecting lung tissue.

Anti-inflammatory potential of microemulsion and pure bullfrog oil in muscle injury

ABSTRACT Introduction: Every day science seeks new ways to treat various diseases through drugs that are efficient and viable. Thus, therapeutic alternatives that meet such demand are targets of study. Microemulsions are one of these new alternatives due to their peculiar pharmacodynamic and pharmacokinetic characteristics. Objective: The aim of this study was to analyze the anti-inflammatory potential of microemulsion and pure bullfrog oil using an experimental model of muscle injury. Methods: Male Swiss mice were divided into three groups: control, microemulsion and pure bullfrog oil. After the pre-treatment, a muscle injury was induced in the animals’ leg and subsequently evaluations were carried out in the horizontal extent of edema and compared between the groups at predetermined times. Following evaluation of muscle injury, dissection of the right gastrocnemius muscles was performed for histological analysis. Results: The microemulsion and pure bullfrog oil showed good anti-inflammatory activity, acting similarly in reducing edema during the first two hours, but without statistical significance from the 3rd to the 24th hour after induction. The histological analysis revealed that the muscle tissue of the animals treated with the microemulsion presented mild cellular infiltrate and little wear of muscle fibers when compared with the muscular tissue of animals treated with the pure bullfrog oil. The histological analysis of the hepatic tissue showed signs of injury in the liver lobes of the pure bullfrog oil group, not observed in the microemulsion group. Conclusion: The microemulsion sho-wed good anti-inflammatory potential in the acute phase of the inflammatory response, reducing the formation of edema and preserving muscle tissue against the occurrence of lesions and without inducing injury in hepatic tissue.