Spontaneous Emulsification Technique Research Articles

Linezolid Loaded Nanoparticle for Topical Administration of Diabetic Foot Treatment: Formulation, In Vitro and Ex Vivo Characterization

The aim of this research work was to develop a topical gel formulation containing nanoparticle to diabetic foot ulcer (DFU). In this respect, the nanoparticle formulations were prepared by using spontaneous emulsification technique. The linezolid (LZD) loaded nanoparticle formulation exhibited an low average particle size (PS) of 195.27±5.42 nm, low polydispersity index (PI) of 0.214±0.019, a high zeta potential (ZP) of 20.57±0.35 and high drug entrapment efficiency (EE) of 99.746±0.021%. In order to enhance topical residence time, the LZD-loaded nanoparticles were dispersed in a gel formulation using Methocel TM K4M (HPMC) and Carbopol® 974 P NF. The formulated gels demonstrated favorable characteristics, including an appropriate pH value, suitable mechanical performance, and desirable viscosity and spreadability for topical administration. All formulations displayed pseudoplastic flow and exhibited typical gel-type mechanical spectra at the specified frequency value. Moreover, the developed formulation achieved sustained drug release as intended for these systems. During the ex vivo drug diffusion studies, 0.007±0.004% of LZD was found in receptor phase, and this was an indicator of local effect. The optimum formulation was stable for 6 months. The initial findings suggest that the formulated topical gel containing LZD-loaded nanoparticles holds promise as an effective drug delivery system for DFU management. However, further comprehensive investigations are required to substantiate this hypothesis.

Quercetin-loaded nanoemulsions prevent Scopolamine-induced neurotoxicity in male rats

Quercetin (QCT) is well-known as a neuroprotective agent due to its antioxidant capacities and reinstating mitochondrial functions. Scopolamine is commonly used as a model to induce Alzheimer's disease (AD-like) symptoms. The current study develops QCT-loaded nanoemulsion (QCT-NE) accompanied by evaluating its neuro-therapeutic effectiveness against SCO-induced neurotoxicity in male rats.The QCT-NE was prepared by the spontaneous emulsification technique and characterized by using particle size, zeta potential, drug loading, in vitro drug release behavior, and stability studies. In vivo studies were done on adult Wistar rats by applying the Morris water maze (MWM) test to study spatial memory and learning. The levels of lipid peroxidation and reduced glutathione were quantitatively determined to reveal the potential mechanism of SCO-induced oxidative stress. Finally, histological studies were performed using staining techniques.The QCT-NE particle size, zeta potential, polydispersity index (PDI), and DL were obtained at 172.4 ± 16.8 nm, -29 ± 0.26 mV, 0.3 ± 0.07, and 81.42 ± 9.14 %, respectively. The QCT and more effectively QCT-NE reduced the elevation of neurobehavioral abnormalities in the MWM test in SCO-exposed rats. The results of oxidative status showed that SCO significantly could increase the LPO and decrease the GSH levels in the rat's brain. However, QCT-NE treatment was more effective than free QCT to inhibit oxidative damage and was well correlated with histopathological findings.Taken together, QCT-NE, compared to QCT, was superior in ameliorating SCO-induced AD-like symptoms due to its better neuroprotective activity and can be considered a novel supplementary therapeutic agent in AD management.

In-Vitro and in-Vivo Evaluation of the Developed Curcumin-Cyclosporine-Loaded Nanoemulgel for the Management of Rheumatoid Arthritis

ABSTRACT Background Topical nanogel-based formulations have shown potential in the management of rheumatoid arthritis (RA). The aim of this research work was to explore the synergistic effect of Curcumin (CUR) and Cyclosporine (CYC) in combination via a topical route for the management of RA. Methods The CUR+CYC loaded nanoemulsion was developed using the spontaneous emulsification technique and was subsequently incorporated into Carbopol® Ultrez 30-NF gel. The effect of the developed formulation on levels of proinflammatory cytokines (IL-6, TNF-α) and anti-inflammatory cytokine (IL-10) was evaluated using lipopolysaccharide (LPS) induced RAW 264.7 cell culture model. The anti-arthritic activity was evaluated in a Complete Freund’s Adjuvant (CFA) induced arthritic rat model. Results The optimized nanoemulgel (CUR + CYC NE gel) exhibited average globule size of 15.32 nm ±2.7 nm, poly-dispersity index of 0.181 ± 0.034 and zeta potential of −16.3 mV ± 0.9 mV. The cumulative drug release from ex-vivo diffusion studies on porcine ear skin was 99.189% ± 1.419% at the of 24 h and 99.177% ± 1.234% at the end of 18 h for CUR and CYC, respectively. The cell culture studies revealed that the formulation was able to significantly lower (p < .001) the levels of IL-6 and TNF-α, inhibited prostaglandin E2 (PGE2) while significantly elevating (p < .001) the levels of anti-inflammatory cytokine (IL-10). The gel was found to be non-irritating and showed the inhibition of paw edema and substantial reduction of arthritic symptoms in an arthritic rat model as compared to commercial and other conventional alternatives. Conclusion This study highlights the potential of the developed nanoemulgel for the management of RA by enhancing the topical permeation of CUR and CYC.

Development and preclinical assessment of nanoemulgel loaded with phytoconstituents for the management of rheumatoid arthritis.

In recent years, natural ingredients have gained importance for therapeutic treatment due to their minimal toxicity. However, the delivery of these phytoconstituents poses a challenge to provide better efficacy. Current research reports the development of nanoemulgel (NEG) loaded with ginger oleoresin (GOR) and lipid guggul extract (LGE) for the management of rheumatoid arthritis (RA). The nanoemulsion (NE) was developed using the spontaneous emulsification technique by the pseudo-ternary method. The optimized nanoemulsion exhibited globule size of 16.08 ± 2.55nm, PDI of 0.187 ± 0.06, and zeta potential of - 22.4 ± 0.31mV. The cumulative release from in vitro diffusion studies at pH 7.4 was about 99.72 ± 3.47%, 57.98 ± 2.11%, and 86.42 ± 5.13% of 6-gingerol, E-guggulsterone, and Z-guggulsterone respectively at the end of 24h. The ex vivo studies on porcine ear skin showed sustained release with 92.8 ± 3.21% for 6-gingerol, 55.61 ± 0.91% for E-guggulsterone, and 84.2 ± 4.22% for Z-guggulsterone released at the end of 24h. The cell culture studies on RAW 264.7 cells indicated a robust inhibition of LPS-induced IL-6 and TNF-α production indicating its efficacy in the management of RA. The preclinical studies on male Wistar rats suggest that the developed NEG exhibited a comparable decrease in paw edema inflammation as compared to the marketed diclofenac sodium gel. These encouraging results demonstrate the potential of the developed nanoemulgel containing combination of GOR and LGE for the management of RA.

The Science of Selecting Excipients for Dermal Self-Emulsifying Drug Delivery Systems.

Self-emulsification is considered a formulation technique that has proven capacity to improve oral drug delivery of poorly soluble drugs by advancing both solubility and bioavailability. The capacity of these formulations to produce emulsions after moderate agitation and dilution by means of water phase addition provides a simplified method to improve delivery of lipophilic drugs, where prolonged drug dissolution in the aqueous environment of the gastro-intestinal (GI) tract is known as the rate-limiting step rendering decreased drug absorption. Additionally, spontaneous emulsification has been reported as an innovative topical drug delivery system that enables successful crossing of mucus membranes as well as skin. The ease of formulation generated by the spontaneous emulsification technique itself is intriguing due to the simplified production procedure and unlimited upscaling possibilities. However, spontaneous emulsification depends solely on selecting excipients that complement each other in order to create a vehicle aimed at optimizing drug delivery. If excipients are not compatible or unable to spontaneously transpire into emulsions once exposed to mild agitation, no self-emulsification will be achieved. Therefore, the generalized view of excipients as inert bystanders facilitating delivery of an active compound cannot be accepted when selecting excipients needed to produce self-emulsifying drug delivery systems (SEDDSs). Hence, this review describes the excipients needed to generate dermal SEDDSs as well as self-double-emulsifying drug delivery systems (SDEDDSs); how to consider combinations that complement the incorporated drug(s); and an overview of using natural excipients as thickening agents and skin penetration enhancers.

Curcumin nanoemulsion counteracts hepatic and cardiac complications associated with high-fat/high-fructose diet in rats.

The relationship between the incidence of cardiovascular abnormalities and non-alcoholic fatty liver disease (NAFLD) has long been postulated. Curcumin (CUR) is a potential anti-atherosclerotic agent but its poor water solubility hinders its pharmacological use. Therefore, the present study aimed to investigate the effect of formulation of CUR nanoemulsion prepared using the spontaneous emulsification technique on high fat high fructose (HFHF)-induced hepatic and cardiac complications. Fifty Wistar rats were divided into five groups. CUR nanoemulsion at doses of 5 and 10mg/kg and conventional powdered CUR at a dose of 50 mg/kg were orally administered daily to rats for two weeks, and compared with normal control and HFHF control. Results revealed that the high dose level of CUR nanoemulsion was superior to conventional CUR in ameliorating the HFHF-induced insulin resistance status and hyperlipidemia, with beneficial impact on rats' recorded electrocardiogram (ECG), serum aspartate aminotransferase (ALT) and alanine aminotransferase (AST) levels, leptin, adiponectin, creatine phosphokinase, lactate dehydrogenase and cardiac troponin-I. In addition, hepatic and cardiac oxidative and nitrosative stresses, oxidative DNA damage and disrupted cellular energy statuses were counteracted. Results were also confirmed by histopathological examination. PRACTICAL APPLICATIONS: The use of curcumin nanoemulsion could be beneficial in combating hepatic and cardiac complications resulting from HFHF diets.

Investigation of Factors Influencing Formation of Nanoemulsion by Spontaneous Emulsification: Impact on Droplet Size, Polydispersity Index, and Stability.

Interest in nanoemulsion technology has increased steadily in recent years for its widespread applications in the delivery of pharmaceuticals, nutraceuticals, and cosmeceuticals. Rational selection of the composition and the preparation method is crucial for developing a stable nanoemulsion system with desired physicochemical characteristics. In the present study, we investigate the influence of intricate factors including composition and preparation conditions that affect characteristic parameters and the stability of the nanoemulsion formation prepared by the spontaneous emulsification method. Octanoic acid, capryol 90, and ethyl oleate were selected to represent oil phases of different carbon–chain lengths. We explored the impact of the addition mode of the oil–Smix phase and aqueous phase, vortexing time, Km (surfactant/cosurfactant) ratio, and the replacement of water by buffers of different pH as an aqueous system. The phase behavior study showed that the Smix phase had a significant impact on the nanoemulsifying ability of the nanoemulsions composed of oil phases of varying carbon-chain lengths. The mode of mixing of the oil–Smix phase to the aqueous phase markedly influenced the mean droplet size and size distribution of the nanoemulsions composed of oil phases as capryol 90. Vortexing time also impacted the mean droplet size and the stability of the generated nanoemulsion system depending on the varying carbon-chain length of the oil phase. The replacement of the water phase by aqueous buffers of pH 1.2, 5.5, 6.8, and 7.4 has altered the mean droplet size and size distribution of the nanoemulsion system. Further, the Km ratio also had a significant influence on the formation of the nanoemulsion system. The findings of this investigation are useful in understanding how the formulation composition and process parameters of the spontaneous emulsification technique are responsible for affecting the physicochemical characteristics and stability of the nanoemulsion system composed of oil of varying carbon-chain (C8-C18) length.

Nanoemulsions Containing Megestrol Acetate: Development, Characterization, and Stability Evaluation.

Many active pharmaceutical ingredients (API) are poorly soluble in water and their low oral bioavailability is a major hindrance to their potential use. Megestrol acetate (MGA) is insoluble in water and its oral absorption is limited and considerably affected by food. Nanoemulsions (NEs) can be used as effective oral drug delivery systems where the hydrophobic API is loaded into the oil phase. In this study, MGA-loaded NEs were prepared based on the spontaneous emulsification technique. The effects of different excipients such as ethanol, Tween 80, Lipoid E80, and medium-chain triglyceride (MCT) on the NEs characterization were investigated. The experimental results indicated that optimum MGA-loaded NEs (F20) were nanometer-sized droplets (166.9 ± 3.0 nm) with negative zeta potential (-12.2 ± 1.1 mV). The effect of polyvinylpyrrolidone (PVP) on characteristic properties of F20 was also evaluated. On the selected NEs, in vitro dissolution tests and stability studies in various mediums and storage conditions were performed. The encapsulation efficiency of NEs were > 99%. The overall droplet size of F20 and PVP-2 (PVP-coated NEs) remained relatively stable as the pH changed from 1.2 to 6.8. It was determined that F20 and PVP-2 remained stable at 4°C until 12 weeks and had higher cytotoxicity on MCF-7 cells. To conclude, droplet size, surface charge, and stability are important properties for NEs to have sufficient effectiveness. In this study, alternative oral NEs of low-solubility drug MGA were developed considering the above features.

Emulsion gels loaded with pancreatic lipase: Preparation from spontaneously made emulsions and assessment of the rheological, microscopic and cargo release properties

Emulsion gels are solidified emulsions, which can be used for delivery of both hydrophilic and lipophilic substances. In this research, at first fish oil-in-water (O/W; 10% w/w) emulsions were prepared through the spontaneous emulsification technique. As emulsifier, a blend of the small-molecule surfactant tween 80, and either low-acyl (LaG) or high-acyl (HaG) gellan was used. For making fully stable (100% stability index) emulsions, a 10-fold higher concentration of LaG than HaG in the emulsion aqueous phase was required. The difference in gellan concentration resulted in a bigger mean drop size, as well as lesser consistency coefficient and yield stress for HaG emulsion than LaG emulsion.Subsequently, the fully stable HaG and LaG emulsions were gelled by CaCl2 addition. LaG emulsion gel was self-supporting and had a dense microstructure (as observed by electron microscopy), whereas HaG emulsion gel was not self-supporting. Loading lipase into the emulsions before ionotropic gelation did not lead to unacceptable acid values for fish oil during the emulsion gels storage.When the lipase-loaded fish O/W emulsion gels were immersed in an acid solution which imitated the gastric fluid (yet without digestive enzymes) oil droplets flocculated (as observed by confocal microscopy). The acid immersion also increased the dynamic moduli of the gels. Lipase was not released into the surrounding acid solution from LaG emulsion gel. A subsequent immersion within an alkaline solution imitating the small intestine fluid (yet without digestive enzymes) reduced the dynamic moduli of both kinds of emulsion gels. The alkaline immersion also caused extensive crack propagation in LaG emulsion gel network, which was found associated with diminished value of tan δ (Gʹʹ/Gʹ) as an index of gel energy dissipation. Lipase was released form LaG emulsion gel into the alkaline solution, however, it took a remarkable period of time to begin.

Formulation, Optimization and in vitro Evaluation of Apremilast Nanoemulgel for Topical Delivery

Background: Psoriasis is an autoimmune-mediated disease characterized marked by thickened plaques and erythematous skin, which can be treated by topical delivery of apremilast. The objective of this study was to prepare hydrogel-based nanoemulsions of apremilast for psoriasis therapy with a view of improving its solubility and permeability. Methods: Nanoemulsion components were chosen based on their solubility. Spontaneous emulsification technique was used to formulate apremilast loaded nanoemulsions D-optimal design was used to optimize the formulations. The optimized formulation was assessed for globule size, Polydispersity Index, percentage transmittance, entrapment efficiency, pH, and Transmission Electron Microscopy The optimized nanoemulsion was converted into gels using carbopol 940 and evaluated for critical parameters like spreadability, viscosity and extrudability. In vitro drug release and ex vivo permeation studies were conducted to understand the release kinetics and extent of permeation. Results: Particle size of the nanoemulsions were found to be in the range of 141 nm- 245nm. Transmission electron microscopic images of the optimised formulation showed spherical particles in the nano range. Optimised formulation exhibited excellent entrapment efficiency of 86% and was found to be thermodynamically stable. In vitro drug release studies of the nanoemulgel using the Franz diffusion cell apparatus appeared to follow zero-order kinetics. Ex vivo permeation through porcine skin showed substantial improvement in flux, permeability coefficient, and drug deposition in the skin in the case of nanoemulgel compared to drug dispersion. Conclusion: The results obtained from the studies demonstrated the immense potential of the developed formulations for topical application in psoriasis.

Polyvinyl alcohol-based electrospun matrix as a delivery system for nanoemulsion containing chalcone against Leishmania (Leishmania) amazonensis

Cutaneous leishmaniasis is a worldwide public health problem. Conventional therapies, in addition to the high cost, have many adverse effects and cases of parasite’s resistance. Chalcones are secondary metabolites precursors in the flavonoid pathway and can be obtained naturally, but with low yield from plant raw material. Thus, the use of synthetic chalcones has been a promising strategy for the development of molecules with leishmanicidal activity. Thus, this work aimed to develop a controlled release system of two synthetic chalcone (trans-chalcones and 3'-(trifluormethyl)-chalcone) using polyvinyl alcohol nanofibers (PVA) as scaffold. The association of chalcones to the nanofibers was made by nanoemulsions (NE) thereof, i.e., a colloidal system on a nanometric scale, which allows compounds with opposite polarities to remain miscible and stable throughout their manipulation. Chalcone nanoemulsions were developed using the spontaneous emulsification technique. The NE were characterized regarding their particle size, polydispersion index (PDI), and zeta potential. The results showed NE with spherical shape, absolute values of zeta potential were higher than 30 mV and homogeneous distribution pattern (PDI < 0.3). Dynamics light scattering (DLS) analysis showed similar hydrodynamic rays, i.e., 180 nm (trans-chalcone NE) and 178 nm (NE containing 3′-(trifluormethyl)-chalcone, in addition to presenting encapsulation efficiency values close to 100 %. Subsequently, the NE were added to a polymeric solution of polyvinyl alcohol (PVA) and processed via the electrospinning technique affording a PVA matrix (15 %, w/v) nanofiber containing the chalcones NE at 1 mg.mL−1. In a follow-up experiment, the skin permeation assay of the PVA matrix-chalcone NE was performed in vitro using Franz type diffusion cells and porcine ear as biological model of study. The results showed that the treatments with the nanofibers containing the chalcone NE were retained mainly in the stratum corneum, while the NE suspensions containing chalcone were retained in the epidermis and dermis. This result is thought to be relevant, since parasites are located mainly in the dermis. Further, in vitro assay against the amastigote form of L. (L) amazonensis, showed IC50 values to trans-chalcone and 3'-(trifluormethyl)-chalcone of 24.42 ± 6.76 μg.mL−1 and 15.36 ± 4.61 μg.mL−1, respectively. In addition to improving the solubility of the compounds tested in culture medium without using organic solvents, chalcones in nano-emulsified form reduced the IC50 to 9.09 ± 1.24 μg.mL−1 (trans-chalcone) and 10.27 ± 2.27 μg.mL−1 (3'-(trifluormethyl)-chalcone) which confirmed the potential of the nanoemulsion containing chalcone for cutaneous leishmaniasis treatment.

Strategies for High Grafting Efficiency of Functional Ligands to Lipid Nanoemulsions for RGD-Mediated Targeting of Tumor Cells In Vitro

Lipid nanoemulsions (LNEs) are promising nanocarriers for delivering high payloads of lipophilic molecules. Nonetheless, the dynamic nature at their aqueous interfaces results in poor surface chemistry and thus ligand functionalization can be challenging. Herein, two independent strategies, postconjugation and preconjugation, were explored to prepare LNEs grafted covalently with model ligands, fluorescein dye and RGD peptide, respectively. Fluorescein was successfully conjugated with high grafting efficiency to an amine-functionalized lipid nanoemulsion (NH2-LNE) as determined by spectrophotometric analysis. First, we formulated NH2-LNEs by a low-energy spontaneous emulsification technique in the presence of oleylamine (OA) within the oily core of the nanodroplets, thus creating primary amine-reactive sites at the oil/water interface. These amines were used to incorporate fluorescein, yielding fluorescent LNEs with grafting efficiencies of 33, 69, and 69% at NH2-LNEs with [OA]oil = 0.18, 0.34, and 0.49 M, respectively. We also developed RGD-labeled LNEs (RGD-LNEs) and evaluated the nanomaterial with model cell lines that overexpress αVβ3 integrins on their surfaces. To this end, we initially synthesized an RGD-Oleate fatty acid-peptide conjugate by solid-phase synthesis. The lipophilic segment of this conjugate readily embedded into the oily core of the LNE, and the hydrophilic head (RGD moiety) was oriented toward the LNE interface. In vitro cytotoxicity and cellular uptake studies were undertaken on different cancer cell lines including HaCaT human umbilical vein endothelial cells (HUVECs), MCF-7, and U-87 MG and compared to uptake experiments with RAW 264.7 macrophages. Confocal imaging and flow cytometry showed that RGD-LNEs were preferentially taken up by all of the tumor cell lines but showed very slight accumulation in RAW macrophages. Unmodified LNE controls did not show any appreciable cellular uptake. This work provides a simple and reliable methodology for the incorporation of multiple ligands on a single surface to facilitate active tumor targeting with LNE-based drug/imaging carriers for theranostic applications.

EFFECT OF CITRONELLA NANO BIOPESTICIDE AGAINST MOSAIC VIRUS AND ITS VECTOR ON PATCHOULI

&lt;p&gt;The mosaic disease declines production and oil quality of patchouli. Antiviral-based citronella oil has been formulated using a spontaneous emulsification technique (nanotechnology). The previous result of the greenhouse trial showed the formula at 1-1.5 % concentrations suppressed the development of virus of about 82.5 %. The field-scale tests is necessary to be performed to validate the effectiveness of citronella nano biopesticide (CNB) against the mosaic virus and its vector on patchouli. The study was conducted in patchouli plantation at two locations (Pandeglang, Banten and Manoko, West Java). The research was arranged in a Randomized Completed Block Design (RCBD) with 6 treatments and 10 replications, each replication consisted of 50 plants. The treatments were formula of CNB at the concentration of (1) 0.5 %, (2) 1 %, (3) 1.5 %, (4) 2 %, (5) insecticide (deltamethrin 0.2 %), and (6) control. The results showed that CNB formula at 1 % concentration with a monthly application effectively suppressed the development of mosaic disease in patchouli plants, and at 2 % concentration to control rolled-leaf aphid. The lowest intensity of mosaic disease (at 1 % concentration) was in Banten (23.12 %) and in West Java (18.35 %), while in control ranged from 26.31-44.94 % (Banten) and 19.60-23.12 % (West Java). Efficacy Index (EI) in Banten ranged from 12.12-48.55 % and in West Java was 6.38-20.63 %. The lowest intensity of aphid attack was showed by insecticide and CNB at 2 % concentration. The EI of CNB was 35.33 % (Banten) and 51.71 % (West Java&lt;strong&gt;) &lt;/strong&gt;respectively.&lt;/p&gt;

Evaluation of the Formation Conditions of a Spontaneous Emulsification Using Porous Silica Particles.

Emulsions are colloid dispersions which are attractive for use as drug carriers due to their simple structure and facile preparation. However, their low physicochemical stability has been problematic. In order to solve this problem, a spontaneous emulsification technique composed of porous silica particles has been developed. In this study, we investigated the conditions for effective formation of protein-encapsulated solid-in-oil-in-water (S/O/W) emulsions using this technique. Porous silica particles having a hydrophilic surface promoted the formation of a fine and uniform emulsion. It was found that the progression of emulsification was affected by electrolytes in aqueous solution. Moreover, it was confirmed that the S/O/W emulsion prepared using this method could successfully encapsulate protein.

Basil Essential Oil and Its Nanoemulsion Mitigate Non Alcoholic Steatohepatitis in Rat Model with Special Reference to Gut Microbiota.

The present research evaluated the protective effect of basil essential oil nanoemulsion (BNO) and its parent basil essential oil (BO) towards steatohepatitis. Chemical composition of BO was assessed followed by formulation into different BNOs using the low energy spontaneous emulsification technique. An ideal formula of BNO was selected among the others based on its ultra-fine particle size (15.42 nm) and physical stability at 25-37°C, which was then tested in steatohepatitis rat model along with BO. Rats were divided into four groups, the first was fed on balanced diet (C), and the other groups were maintained on high fructose saturated fat diet deficient in choline to induce steatohepatitis, one of such groups served as control steatohepatitis (SC), the other groups received daily oral dose of BO and BNO, respectively. Microbiota (Firmicutes and Bacteroidetes) were counted in colon content and their ratio (F/B) was calculated. Liver fat, plasma lipid profile, plama interlukin-6, plasma lipopolysaccharides and plasma and colon content of lipocaline were assessed with histopathological examination of liver and colon. Results showed that the major volatile components of BO were linalool (60.9 %), eugenol (5.1 %) and eucalyptol (9.5%). SC group exhibited significant increase in liver lipids, plasma triglycerides, total cholesterol (TC), low density lipoprotein cholesterol and significant reduction in high density lipoprotein-cholesterol (HDL-C) compared to C group. Significant increase in plasma TC/HDL-C, interlukin-6, and lipocaline and F/B ratio and lipocaline in colon content were demonstrated in SC group without changes in plasma lipopolysaccharides compared to C. Histopathology of SC group showed liver fatty degeneration and fibroblasts activation while the colon demonstrated erosion and mucosal epithelium detachment. Treatment with either BNO or BO showed improvement compared to SC group. BNO was superior in reducing F/B ratio, liver lipids and histopathological changes. BO was more efficient in reducing TC, triglycerides and low density lipoprotein cholesterol. It is concluded that BO and BNO reduced the progression of nonalcoholic steatohepatitis in rat model. Gut microbiota in relation to steatohepatitis and related new therapies needs further investigations.

Enrichment of Eucalyptus oil nanoemulsion by micellar nanotechnology: transdermal analgesic activity using hot plate test in rats\u2019 assay

Eucalyptus globulus is an aromatic medicinal plant which known for its 1,8-cineole main pharmacological constituent exhibits as natural analgesic agent. Eucalyptus globulus-loaded micellar nanoparticle was developed via spontaneous emulsification technique and further evaluation for its analgesic efficacy study, in vivo analgesic activity assay in rats. The nanoemulsion system containing Eucalyptus-micelles was optimized at different surfactant types (Tween 40, 60 and 80) and concentrations (3.0, 6.0, 9.0, 12.0, 15.0, and 18.0 wt. %). These formulations were characterized by thermodynamically stability, viscosity, micelles particle size, pH, and morphology structure. The spontaneous emulsification technique offered a greener micelles formation in nanoemulsion system by slowly titrated of organic phase, containing Eucalyptus globulus (active compound), grape seed oil (carrier oil) and hydrophilic surfactant into aqueous phase, and continuously stirred for 30 min to form a homogeneity solution. The characterizations evaluation revealed an optimized formulation with Tween 40 surfactant type at 9.0 wt. % of surfactant concentration promoted the most thermodynamic stability, smaller micelles particle size (d = 17.13 ± 0.035 nm) formed with spherical shape morphological structure, and suitable in viscosity (≈2.3 cP) and pH value (6.57) for transdermal purpose. The in vivo analgesic activity assay of optimized emulsion showed that the transdermal administration of micellar nanoparticle of Eucalyptus globulus on fore and hind limb of rats, possessed the central and peripheral analgesic effects by prolonged the rats pain responses towards the heat stimulus after being put on top of hot plate (55 °C), with longest time responses, 40.75 s at 60 min after treatment administration. Thus, this study demonstrated that micellar nanoparticle of Eucalyptus globulus formed in nanoemulsion system could be promising as an efficient transdermal nanocarrier for the analgesic therapy alternative.

Anticancer and antibacterial effects of a clove bud essential oil-based nanoscale emulsion system.

Background and purposeThe essential oil derived from clove buds (Syzygium aromaticum) has been used as a chemopreventive agent in Ayurvedic medicine. The antiviral, antibacterial, and anticancer properties of its chemo-skeleton have motivated this study to explore its efficacy in pharmaceutics.MethodsNanoscale-based emulsions were prepared by employing a spontaneous emulsification technique through self-assembly using varying concentrations of Tween 20 and Tween 80 surfactants. Their physicochemical properties and stability were studied in order to choose an optimum formulation which was clear and stable. The cytotoxicity of the stable oil-based emulsion system was evaluated using MTT assay, colony formation assay, and Annexin V-FITC assay against the thyroid cancer cell line (HTh-7).ResultsAll three methods verified apoptosis and reduction in cancer cell proliferation, making the formulation a promising candidate as an alternative cancer drug. The oil-based emulsion system was also tested for its antibacterial properties against Staphylococcus aureus. Membrane permeability studies proved its efficacy to permeate through cell membrane, thereby increasing the leakage of cytoplasmic contents.ConclusionMany current treatments for cancers are aggressive yet ineffective. This study positions the clove bud-based nanoscale emulsion as a suitable candidate for further in vivo studies and trials as a cancer drug.

Rapamycin-Loaded, CapryolTM 90 and Oleic Acid Mediated Nanoemulsions: Formulation Development, Characterization and Toxicity Assessment.

This study was planned to explore the capability of nanoemulsions (NEs) consisting of CapryolTM 90 and oleic acid for the delivery of rapamycin (RAP). Permeability and cytotoxicity of RAP-loaded NEs were also inspected. Pseudo-ternary phase diagrams were created with oleic acid and CapryolTM 90 (as oil phase) and four surfactants and co-surfactants at various weight ratios (Rsm). Selected NEs from O/W region on the phase diagrams with the drug concentration of 1 mg/mL, were prepared via the spontaneous emulsification technique, characterized for particle size and subjected to stability tests at various temperatures over 9-12 months. Cumulative drug release was determined for a period of 48 h using a dialysis sac. The assay of RAP was determined using HPLC technique. Cytotoxicity of NEs was evaluated by MTT assay on breast cancer cell line, namely SKBR-3. The permeability of RAP-loaded NEs across Caco-2 monolayers was assessed by measurement of TEER (transepithelial electrical resistance) value. The intracellular uptake of coumarin 6-loaded NEs by SKBR-3 cells was also investigated using florescence microscopy. NEs containing oleic acid/Tween 20/propylene glycol, CapryolTM 90/Tween 20/iso-propanol, and CapryolTM 90/Cremophor® RH40/Transcutol® P showed more cytotoxicity and permeability compared with the RAP methanolic solution. The minimum toxic concentration of RAP in NE formulations was found to be 7.5 µg/mL. The highest intracellular uptake was observed for the NE composed of CapryolTM 90/Tween 20/iso-propanol which was in consistent with the results obtained from cytotoxicity and permeability tests. The overall results implicated that this novel carrier was effective for enhancing RAP permeation in Caco-2 cell membrane along with enhancement of cytotoxicity.

Lippia gracilis Schauer essential oil nanoformulation prototype for the control of Thielaviopis paradoxa

The goal of the study was to evaluate the antifugal effect of essentials oils from the Lippia gracilis genotypes with main components of thymol and carvacrol and essential oil emulsions on Thielaviopis paradoxa. The essential oil was extracted from the genotypes by hydrodistillation. The nanoemulsions were obtained from the spontaneous emulsification technique. The nanoemulsions obtained where characterized by particle size, zeta potential and polydispersity. For the fungal activity bioasssays in vitro, the essential oils, majority components and nanoemulsions in different concentrations were added to potato-dextrose-agar (PDA) medium. Each plate was inoculated placing at the center a 7 mm disk containing T. paradoxa mycelium. For the in vivo assay, for the plant infections a 2 × 106 CFU (colony forming unit) suspension in was use in PDA medium. After 7 days, the emulsions were pulverized with minimal inhibitory concentrations and testemonies. The ecophysiology parameters (photosynthesis; stomatal conductance, transpiration and vapor pressure deficit between plant and air) in 30 coconut trees inoculated with T. paradoxa was analyzed. The L. gracilis essential oil has antifungal potential for the control of coconut tree stem-bleeding, since the T. paradoxa was inhibited in all concentrations used. The genotype LGRA109 essential oil presented MIC of 0.23 mg/mL, while the LGRA106 presented MIC of 0.82 mg/mL. The essential oil nanoemulsions presented high efficiency in disease control (MIC 0.6 mg/mL) and low phytotoxicity; therefore representing a good alternative for the efficient control of T. paradoxa in coconut tree.

Development of a Novel Spontaneous Emulsification Method for Peptide Delivery Using Porous Silica Particles.

A new emulsification technique using porous silica particles was studied as a facile and instantaneous formation method for thermodynamically unstable emulsions. In this study, oil encapsulated in silica particles was released instantly upon the addition of a phosphate buffer, forming an oil-in-water (O/W) emulsion. Emulsion formation was inhibited or promoted using lipophilic or hydrophilic surfactant additives, respectively. We concluded that this phenomenon is affected by the wettability of the soybean oil on the silica surface, which is controlled by the surfactant. We prepared submicron size emulsions in a simple method involving the addition of the silica particles to an aqueous solution. This spontaneous emulsification technique could be applied to the formation of solid-in-oil-in-water (S/O/W) emulsions for oral delivery of hydrophilic peptide medicine.