Nanoemulsion Formulation Research Articles

Isolation of Bioactive Compounds (Carotenoids, Tocopherols, and Tocotrienols) from Calendula Officinalis L., and Their Interaction with Proteins and Oils in Nanoemulsion Formulation.

Calendula officinalis L. has numerous health-promoting properties due to the presence of a large number of lipophilic compounds. Their effective delivery to the body requires the use of an appropriate technique such as emulsification. So, the main purpose of this study was to understand how the profile of lipophilic compounds from pot marigold and the pro-health potential are shaped by different types of protein, oil, and drying techniques in o/w nanoemulsion. To obtain this, the profiles of carotenoid compounds and tocols were measured. Additionally, antioxidant potential and the ability to inhibit α-amylase and α-glucosidase were measured. Pea protein emulsion exhibited a higher final content of carotenoid compounds (23.72-39.74 mg/100 g), whereas those with whey protein had stronger α-amylase inhibition (487.70 mg/mL). The predominant compounds in the studied nanoemulsions were β-carotene (between 19% and 40%), followed by α-tocopherol/γ-tocopherol. The type of proteins shaped the health-promoting properties and determined the content of health-promoting compounds.

Characterization and In vitro Release & In vivo Behavior Study of Self-Assembled Nano-Emulsion in XiaoYao Pill for Enhanced Drug Delivery.

Traditional Chinese medicine formulations often contain hydrophobic components with limited solubility and stability, leading to low oral bioavailability. Self-assembled nanoparticles (SANs) have shown promise in enhancing oral bioavailability of these components. However, whether un-decocted Chinese herbal pellets can generate SANs and the impact of SANs formed by multiple components on pharmacokinetic parameters remains unexplored. In this study, single-factor approach was employed to determine the optimal separation method of nano-emulsion phase of XiaoYao pill (N-XY). Morphological and particle size analyses confirmed the nanoscale nature of N-XY. High-performance liquid chromatography (HPLC) fingerprint analysis was conducted to compare the distribution of active ingredients among three different phases of XiaoYao pill (XY pill). In vitro release studies were performed to evaluate the release mechanism of four ingredients from N-XY. Additionally, in vivo pharmacokinetics and tissue distribution behaviors were investigated in rats. N-XY exhibited uniform and stable characteristics as a water-in-oil (O/W) nano-emulsion. Fingerprint analysis identified 25 characteristic peaks and 8 key ingredients in N-XY, with the highest peak areas. In vitro release studies showed a sustained release behavior of N-XY. The pharmacokinetics study showed that the ferulic acid of N-XY had a 1.37-fold higher AUC, 1.44-fold lower Vd/F, 1.39-fold lower CL/F, and a prolonged t1/2 than A-XY, indicating enhanced bioavailability due to reduced elimination. Furthermore, the tissue distribution revealed that the levels of paeoniflorin and ferulic acid from N-XY significantly increased in liver, spleen, lungs, uterus and ovaries, exhibiting targeting characteristics. This study comprehensively explored the formation, characterization, and pharmacokinetics of nano-emulsion in XY pill, introducing novel perspectives and initiating preliminary research on potential SANs in un-decocted traditional Chinese medicine formulations. It also emphasized the importance of enhancing pharmacokinetics of hydrophobic components in Chinese herbal formulations and laid the foundation for future nano-formulation research for XY pill.

Ex Vivo Irritation Evaluation of a Novel Brimonidine Nanoemulsion Using the Hen's Egg Test on Chorioallantoic Membrane (HET-CAM).

The hen's egg test on chorioallantoic membrane (HET-CAM) assay is a cost-effective and well-validated, non-animal-based ex vivo method for evaluating the irritant potential and eye toxicity of substances. A colloidal dispersion of a surfactant and a cosurfactant with a nanosize range is called a nanoemulsion (NE), which is formed by mixing immiscible liquids and stabilized by surfactants. Patients with glaucoma are commonly prescribed Brimonidine (BR), an alpha-2 adrenergic agonist, to lower their intraocular pressure. In this study, surfactant-cosurfactant blends were prepared by mixing Tween 80 (surfactant) and propylene glycol (cosurfactant) in a 4:1(v/v) ratio. Triacetin served as the oil phase, while deionized water was used as the aqueous phase. Using the drop method, a range of NE formulations (F1, F2, F3, FB1, FB2, and FB3) were developed and subsequently evaluated for their potential to irritate, and then the results were compared to those of a commercially available BR eye drop formulation. According to the average cumulative HET-CAM test scores (IS), from excipients, propylene glycol caused moderate irritation by causing slight damage to blood vessels. The formulations FB1 and F1 were found to have the highest level of irritation among other formulations in the investigation, recording 1.05 ±0.07 and 1.2 ±0.10, respectively. All other NE formulations exhibited non-irritating potential, as confirmed by the HET-CAM test, and were comparable to the marketed BR eye drop formulation. The NE formulations created for BR were determined to be safe and non-irritating. The findings indicate that the prepared NE could be a beneficial solution for addressing problems with conventional eye drops and delivering BR effectively to the eyes.

Advances in Nanoemulsion Formulations for Enhanced Drug Delivery: A Comprehensive Review

Abstract: Nanoemulsions (NEs) have emerged as adaptable medication delivery methods with extensive applications in both pharmaceuticals and cosmeceuticals. This comprehensive review delves into the myriad advantages of NEs, ranging from enhanced drug absorption to providing stability for chemically unstable compounds. NEs exhibit promising potential as substitutes for liposomes, offering improved performance in various formulations. In the realm of cosmeceuticals, NEs play a pivotal role in enabling rapid skin penetration while maintaining skin moisturization, thanks to their visually appealing coating. Their ability to deliver active ingredients efficiently makes them an attractive option for cosmetic formulations seeking enhanced efficacy. Moreover, this review sheds light on the involvement of NEs in antimicrobial therapies and gene therapy, underscoring their versatility and potential in addressing diverse medical challenges. Additionally, NEs find applications in commercial disinfectants, further highlighting their broad utility beyond traditional drug delivery systems. Furthermore, the review discusses innovative strategies, such as solid self-nanoemulsifying drug delivery systems (S-SNEDDS), which offer advantages in terms of stability, drug loading capacity, and patient compliance. By exploring these strategies, researchers aim to overcome existing limitations and optimize the therapeutic potential of NEs.

Development of Nanoemulsion Formulation Containing Ylang Ylang Essential Oil for Topical Applications, Evaluation of In Vitro Cytotoxicity and ADMET Profile

Ultraviolet (UV) rays damage DNA, causing adverse effects such as photoaging and cancer on the skin. For the well-being of individuals, there is a need to develop innovative skin products with high effectiveness using protective and therapeutic agents. In this context, in our study, A nanoemulsion (NE) formulation containing Ylang-ylang essential oil (YO), which has many biological active properties such as antimicrobial, antioxidant, anti-inflammatory, and anticancer, was produced by the ultrasonic emulsification method and characterized. The thermodynamic stability was evaluated, and its in vitro release profile determined the dialysis membrane technique. The cytotoxic effect of YO-NE was examined with the in vitro method on the HacaT cell line using the MTT method and in silico method using the ADMET profile. Dynamic light scattering (DLS) results showed that the average droplet size of the YO-NE formulation was 184.1±2.307 nm, the polydispersity index (PdI) was 0.151±0.006, and the Zeta potential (ζ) -10.8 ± 0.400 mV. As a result of in vitro release studies, it was observed that 99.98± 1.00% of YO release from NE occurred within 5 hours. Based on the thermodynamic stability test results, it was determined that the developed formulation did not show sedimentation or phase separation. Also, it remained stable under three different storage conditions in the three-month stability test. Cytotoxicity results revealed that the YO-NE formulation was safe. All the results indicated that the YO-NE formulation might be considered a non-toxic product candidate with physicochemical properties suitable for topical use.

The Use of Lemongrass/Lemon Blend Essential Oil Nanoemulsions in Chewy Candy Formulations and Its Evaluation Against Streptococcus mutans and Porphyromonas gingivalis

Lemongrass (Lg) and lemon (Lm) essential oils (EOs) mainly contain citral and limonene, respectively, which are not limited to flavor for food but can also provide antibacterial activity. The information on the enrichment of these combined EOs in chewy candy products has never been documented yet. This study aimed to evaluate the physical-chemical properties and volatile compound of nanoemulsion formulations of Lg/Lm blend EOs as well as the physical-chemical properties, total plate count (TPC), and sensory acceptability of chewy candy formulated with nanoemulsions. Antibacterial evaluations against Streptococcus mutans and Porphyromonas gingivalis were performed for both nanoemulsion and chewy candy. Blending Lg and Lm EOs at ratios of Lg1.85 : Lm2.25 and Lg3.65 : Lm0.45 showed a broadly different constituent of the volatile compound than the ratio with the least amount of Lg (Lg0.05 : Lm4.05) but all the formulations possessed eugenol, β-Bisabolene, and caryophyllene oxide. The combination of Lg1.85 : Lm2.25 produced the nanoemulsion with the lowest particle size but lower zeta potential and emulsion stability. At this ratio, the significant inhibitory activity of the nanoemulsion was found to be 92.40% and 84.14% for S. mutans and P. gingivalis, respectively (p < 0.05). Its application in chewy candy formulation also resulted in the highest inhibitory activity against S. mutans (85%) and P. gingivalis (77.20%). Chewy candy formulated with Lg1.85 : Lm2.25 nanoemulsion was also found to have the lowest TPC of 3.72 log CFU/g. The overall acceptability of the chewy candy was around neutral (3.77) by panelists for its higher color score (4.73) and aroma (4.37) in average to the other samples. This study discovered the potential of Lg/Lm blend EOs nanoemulsion for enhancing the antibacterial effect and improving the texture and color of chewy candy.

Optimization of Polylactide-Co-Glycolide-Rifampicin Nanoparticle Synthesis, In Vitro Study of Mucoadhesion and Drug Release.

Despite the large number of works on the synthesis of polylactide-co-glycolide (PLGA) nanoparticles (NP) loaded with antituberculosis drugs, the data on the influence of various factors on the final characteristics of the complexes are quite contradictory. In the present study, a comprehensive analysis of the effect of multiple factors, including the molecular weight of PLGA, on the size and stability of nanoparticles, as well as the loading efficiency and release of the antituberculosis drug rifampicin (RIF), was carried out. Emulsification was carried out using different surfactants (polyvinyl alcohol, Tween 80 and Pluronic F127), different aqueous-to-organic phase ratios, and different solvents (dichloromethane, dimethyl sulfoxide, ethyl acetate). In this research, the PLGA nanoemulsion formation process was accompanied by ultrasonic dispersion, at different frequencies and durations of homogenization. The use of the central composite design method made it possible to select optimal conditions for the preparation of PLGA-RIF NPs (particle size 223 ± 2 nm, loading efficiency 67 ± 1%, nanoparticles yield 47 ± 2%). The release of rifampicin from PLGA NPs was studied for the first time using the flow cell method and vertical diffusion method on Franz cells at different pH levels, simulating the gastrointestinal tract. For the purpose of the possible inhalation administration of rifampicin immobilized in PLGA NPs, their mucoadhesion to mucin was studied, and a high degree of adhesion of polymeric nanoparticles to the mucosa was shown (more than 40% within 4 h). In the example of strain H37Rv in vitro, the sensitivity of Mycobacterium tuberculosis to PLGA-RIF NPs was proven by the complete inhibition of their growth.

Development and evaluation of methotrexate-loaded nanoemulsion formulation for topical treatment of psoriasis.

Psoriasis is a chronic inflammatory disease that is becoming widespread and is associated with many kinds of additional severe diseases. The present study aimed to develop a methotrexate-loaded almond oil-based nanoemulsion formulation (MTX NE) for topical administration. The drug-loaded nanoemulsion formulation was prepared by high shear homogenization technique. The formulation's stability, as well as other physical and chemical characteristics, including entrapment effectiveness, drug release kinetics, skin permeability, skin irritation, and in vivo evaluation of the optimized formulation, was assessed. Additionally, imiquimod-induced psoriasis in rats was employed to investigate the efficacy of MTX NE against skin disorders. The MTX NE formulation was developed with a particle size of 18.74 ± 9.748nm, a polydispersity index (PDI) of 0.198 ± 0.01, and an average entrapment efficiency of 79.65 ± 3.84%. The release kinetics model estimates 81.08% drug release at pH 5.5 after 24h. The major layers of the skin, the epidermis, and dermis were successfully fluidized by the optimized MTX NE formulation, as shown by FTIR results, most likely enhancing drug retention and permeability. However, since Tween 80 and PEG 400 are well-known penetration enhancers, their application greatly accelerates these effects. Permeation data indicate that after 24h, methotrexate was released from the nano-emulsion at 76.83 ± 4.98g/cm2 with a flux rate of 2.385 ± 0.61µg/cm2/h. The in vivo study conducted on rabbit skin showed that the enhanced skin penetration of the prepared MTX-loaded nanoemulsion formulation does not cause any structural modifications in the inter-cellular lipid layers of the stratum corneum. Rabbits used in the in vivo anti-psoriatic investigation demonstrated that MTX NE produced a 95% reduction in PASI. The pharmacokinetic profile revealed that the Cmax, Tmax, and t1/2 values were 8.63µg/mL, 12.5h, and 17.77 ± 2.21h, respectively. These findings suggest that the formulation MTX NE is effective in treating psoriasis and may reduce psoriasis symptoms.

Utilization of nanoemulsion to enhance antibacterial capacity of Zanthoxylum bungeanum pericarp essential oils against foodborne pathogenic bacteria

Essential oils (EOs) have promising applications as natural and environmentally friendly antimicrobial agents. Preparation of EOs into emulsions can improve the properties of EOs. The aim of this study was to investigate the mechanism underlying the antibacterial effects of Zanthoxylum bungeanum pericarp essential oil (ZBEO) against Staphylococcus aureus, Escherichia coli, Bacillus cereus, and Shigella boydii, and to determine the effect of nanoemulsions on antibacterial activity. ZBEO efficiently suppressed foodborne pathogenic bacteria, as determined by the minimum inhibitory concentration and survival rate. Based on analyses of bacterial morphology, the degree of leakage of proteins and nucleic acids, and lactate dehydrogenase activity, we found that ZBEO disrupted the integrity and permeability of cell membranes, resulting in the leakage of cellular contents and apoptotic cell death. Prepared ZBEO nanoemulsion (ZBEO-NE) had good dispersion and stability. Both ZBEO and ZBEO-NE inhibited the growth of S. aureus, E. coli, B. cereus, and Sh. boydii in mutton broth, and ZBEO-NE exhibited better inhibitory effects than those of ZBEO. These findings suggest that preparing EOs in the form of nanoemulsions improves the antibacterial properties of EOs, making them suitable for food applications.

Andiroba Oil (Carapa guianensis) and ginger extract (Zingiber officinale)-loaded nanoemulsion: Elaboration, characterization, antioxidant activity, and cell viability investigation

This study employs nanoengineering techniques to enhance the aqueous compatibility of andiroba oil and ginger extract, both renowned for their phytotherapeutic properties, aiming at biomedical applications. The natural products were characterized using HPLC-MS, CG-MS, UV-Vis, and FTIR. Optimal surfactant concentrations (lecithin and Tween® 20) were established to produce more biocompatible and stable oil-in-water nanoemulsions. The nanodroplets were characterized at multiple hierarchical levels, including their shape (spherical), size (∼130 nm), polydispersity (PDI = 0.20), and surfactant interfacial film architecture through a cross-analysis of DLS, ELS, SAXS, TGA, UV-Vis, and FTIR. Zetametry and DLS were used to investigate the mechanism of colloidal stabilization of the nanodrops and evaluate the stability of the nanoemulsion under varying pH and ionic strength conditions during storage at different temperatures. The quantification of phenol and flavonoids, coupled with assessments of antioxidant activity, demonstrated an additive effect, indicating that both andiroba oil and ginger extract retain their significant antioxidant efficacy within the nanoemulsion formulation. The effect of the nanoformulation on the viability and morphology of mouse fibroblast NIH-3T3 was also examined. The IC50 of the nanoemulsion at 48 h (765 μg/ml) is higher than the values reported for other nanoemulsions of andiroba oil or ginger extract. This innovative nanoemulsion system combining andiroba oil and ginger extract offers the potential for additive or synergistic effects while exerting minimal impact on healthy cell viability.

Enhancing bioactivity of Callistemon citrinus (Curtis) essential oil through novel nanoemulsion formulation

The main focus of this study was to create a stable and efficient nanoemulsion (NE) using Callistemon citrinus essential oil (EO). Various factors affecting the NE’s stability were optimized including oil %, Tween 80%, time of sonication, and its accelerated stability was examined. The research also considered the antibacterial, antifungal, and larvicidal effects of the optimized NE (B10). The optimum NE stood out for its stability, featuring a particle size of 33.15 ± 0.32 nm. Analysis via IR spectroscopy confirmed successful EO encapsulation in B10. The formulation remained stable for six months, with B10 showing significantly higher antibacterial and antifungal potency compared to the pure oil. When samples were subjected to tests against Fusarium oxysporum, B10 exhibited a MIC value of 62.5 mg/mL, whereas the pure oil showed a MIC value of 250 mg/mL. This indicates that the B10 formulation was 50 times more effective than the EO. In terms of antibacterial activity against Escherichia coli, the MIC value was 0.256 mg/mL for B10 and 4 mg/mL for the EO. Also, pure oil and B10 displayed larvicidal effects against Chilo suppressalis (Walker) larvae, with B10 eliminating 95.2% of larvae in 48 h. Overall, stable and optimum C. citrinus NE with its strong antimicrobial qualities, shows promise as an effective fungicide and insecticide.

Formulation, characterization and evaluation of anti-breast cancer activity of 2-carene nanoemulsion; in silico, in vitro and in vivo study

Natural phytocompounds extensively used in several therapeutic regimes, biomedical and pharmaceutical applications and their nanoparticles have become extremely essential in field of nanotechnology to treat various types of cancer. The main objective of this study demonstrates the antitumor activities of pure terpene compound 2-carene and its nanoemulsion against in vitro and in vivo breast cancer model. In silico and molecular dynamic simulation study was performed against breast cancer receptors to evaluate anticancer activity of 2-carene. Nanoemulsions were prepared by isolating monoterpenoid compound from Catharanthus roseus essential oil and anti-cancer effects of 2-carene mediated nanoemulsion were evaluated. Gas chromatography-mass spectrometry analysis explicated 2-carene was the active terpene compound and nuclear magnetic resonance spectroscopy was performed to confirm structural identification of this compound. Nanoemulsion was characterized by characteristic techniques of scanning electron microscopy, Fourier transform infrared spectroscopy, size determination and dynamic light scattering. Cytotoxicity, antioxidant activity, apoptosis and histopathological examination were used to identify bioactive qualities of prepared nanoemulsion formulation. Effective cytotoxicity of nanoemulsion was observed 54.31% against MDA-MB-231 cancer cell lines. Tumor features and apoptotic activity was also assessed and the results of study strongly revealed that bio-synthesized nanoemulsion show significant apoptotic and anti-tumor potential against DMBA induced breast cancer. The antioxidant enzymes and serum inflammatory markers were significantly improved by nanoemulsion treatment in cancer induced rat model. In histopathological evaluation, the nanoemulsion formulations markedly show signs of improvement in cancer induced tissues. Results demonstrated that 2-carene and its synthesized nanoemulsion exhibit remarkable anticancer potential against DMBA induced cancer.

Development and evaluation of nanoemulsion gel loaded with bioactive extract of Cucumis melo var. agrestis: A novel approach for enhanced skin permeability and antifungal activity

The utilization of phytoconstituents in skin care products has emerged as a notable trend due to their recognized safety and therapeutic efficacy. However, the challenge lies in improving the effective delivery of phytoconstituents to specific tissues, primarily attributed to their poor solubility and low permeability. This study endeavors to address this challenge by developing, optimizing and characterizing Cucumis melo var. agrestis (CME) extract loaded nanoemulsion gel (CME-NEG), aiming to enhance the skin permeability and antifungal activity. Herein, nanoemulsions encapsulating the plant extract were prepared using ultrasonication technique and were characterized for droplet size, zeta potential, polydispersity index (PDI) and entrapment efficiency. Further, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) analysis were conducted to characterize the optimized CME extract loaded nanoemulsion (CME-NE 3) formulation. The optimized formulation was blended with Carbopol 940 gel to develop CME-NEG, which was evaluated for release kinetics, in vitro permeation and in vitro antifungal activity. High performance liquid chromatography (HPLC) analysis confirmed the presence of gallic acid, chlorogenic acid, 4-Hydroxy benzoic acid (HB acid), kaempferol, caffeic acid and quercetin. Findings of 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay showed that the ethanolic extract had highest antioxidant activity (88.88 %). The optimized formulation displayed smooth spherical nanodroplets with size of 175.5 ± 1.56 nm, zeta potential of −21.5 ± 0.12 mV, PDI of 0.192 ± 0.06, and highest entrapment efficiency (EE) of 91.35 ± 1.65 %. The release profile of CME-NE exhibited a controlled release characteristic and the release kinetic mechanism was best described by the Korsmeyer-Peppas (Kp) model. In a 24 h permeation study, it was observed that the in vitro permeation of CME-NEG was 58.63 %, significantly higher than that of CME extract loaded plain gel (CME-PG) with an enhancement ratio of 2.12. The prepared CME-NEG formulation also presented enhanced antifungal activity as compared to pure CME extract. In conclusion, the designed CME-NEG offers a promising topical drug delivery system with enhanced skin permeability and antifungal activity.

Optimization and characterization of essential oil-loaded nanoemulsions for enhanced antiviral activity against tobacco mosaic virus (TMV) in tobacco plants

Tobacco mosaic virus (TMV) is a significant viral pathogen that infects various economically important crops, including tobacco, tomatoes, and cucumbers. The current study aimed to develop and optimize essential oil-loaded nanoemulsions using Response Surface Methodology (RSM) to enhance antiviral activity against TMV in Nicotiana tabacum plants. Essential oils from tea tree (Melaleuca alternifolia) and lime peel (Citrus latifolia) were chemically analyzed using gas chromatography-mass spectrometry (GC-MS). Terpinene-4-ol (70.92 %) and trans-isolimonene (33.82 %) were identified as the major components in tea tree and lime peel oils, respectively. RSM was used to optimize the nanoemulsion formulations based on oil concentration, hydrophilic-lipophilic balance (HLB), and stirring speed. Optimal tea tree oil (TTEO) and lime peel oil (LPEO) nanoemulsions were developed and characterized using Fourier-transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and particle size analysis. The antiviral efficacy of the optimized formulations was tested against TMV using RT-qPCR analysis and physiological measurements. TTEO nanoemulsions significantly inhibited TMV expression by 89.78 %, while LPEO nanoemulsions achieved a 42.22 % reduction after six days. Additionally, these essential oils (EOs) enhanced plant defense mechanisms by modulating the activities of phenylalanine ammonia-lyase (PAL) and peroxidase (POD), along with maintaining chlorophyll levels. This study demonstrates that TTEO and LPEO nanoemulsions offer promising eco-friendly antiviral agents for TMV management in tobacco plants. The research underscores the potential of essential oil-based formulations in mitigating viral infections in crops, providing a sustainable alternative to synthetic chemicals.

Formulation of catechin hydrate nanoemulsion for fortification of yogurt

Formulation of catechin hydrate nanoemulsion for fortification of yogurt

Enhancing the efficacy of fluconazole against Leishmania major: Formulation and evaluation of FLZ-nanoemulsions for topical delivery

BackgroundCutaneous Leishmaniasis (CL) remains a significant public health concern, particularly in the tropical and subtropical regions. Present treatment options for CL such as Fluconazole (FLZ) face limitations, including low solubility and bioavailability. This study aimed to address these challenges by investigating the use of nano-emulsions (NEs) to enhance the efficacy of FLZ against Leishmania major(L.major). Materials and methodsFLZ-NEs were formulated with oleic acid, Tween-20, and ethanol using low-energy emulsification at various surfactant/co-surfactant ratios. Subsequently, a comprehensive analysis was conducted to assess the physicochemical characteristics of the samples. This analysis encompassed stability, zeta potential, pH, viscosity, refractive index, and droplet size. We then studied the anti-parasitic properties of these optimized FLZ-NEs both in vitro and in vivo. ResultsThe selected nano-emulsion (NE) formulation (2 % oleic acid, 20 % Tween 20, 10 % ethyl alcohol) showcased desirable properties like small droplet size (10.51 ± 0.24 nm), low dispersity (0.19 ± 0.03), and zeta potential value (- 0.41 ± 0.17 mV), key for stability and targeted drug delivery. This optimal formulation translated into remarkable efficacy. In vitro, FLZ-NEs demonstrated a threefold increase in their ability to combat promastigotes and a remarkable thirtyfold increase in their ability to combat amastigotes. Additionally, they demonstrated a ninefold advantage in their ability to specifically target parasites within infected macrophages, thereby attacking the infection site. These promising in vitro results translated into improved outcomes in vivo. Compared to other chemicals studied, FLZ-NE-treated mice showed decreased disease severity, weight growth, and quicker ulcer healing. It was further supported by histopathological research, which showed reduced tissue damage linked to Leishmania infection. ConclusionThese findings show the potential of nanotechnology-based drug delivery in improving anti-leishmanial treatment.

Green synthesis of zinc oxide nanoparticles (ZnO-NPs) by Streptomyces baarnensis and its active metabolite (Ka): a promising combination against multidrug-resistant ESKAPE pathogens and cytotoxicity

Various eco-friendly techniques are being researched for synthesizing ZnO-NPs, known for their bioactivity. This study aimed at biosynthesizing ZnO-NPs using Streptomyces baarnensis MH-133, characterizing their physicochemical properties, investigating antibacterial activity, and enhancement of their efficacy by combining them with a water-insoluble active compound (Ka) in a nanoemulsion form. Ka is a pure compound of 9-Ethyl-1,4,6,9,10-pentahydroxy-7,8,9,10-tetrahydrotetracene-5,12-dione obtained previously from our strain of Streptomyces baarnensis MH-133. Biosynthesized ZnO-NPs employing Streptomyces baarnensis MH-133 filtrate and zinc sulfate (ZnSO4.7H2O) as a precursor were purified and characterized by physicochemical investigation. High-resolution-transmission electron microscopy (HR-TEM) verified the effective biosynthesis of ZnO-NPs (size < 12 nm), whereas dynamic light scattering (DLS) analysis showed an average size of 17.5 nm. X-ray diffraction (XRD) exhibited characteristic diffraction patterns that confirmed crystalline structure. ZnO-NPs efficiently inhibited both Gram-positive and Gram-negative bacteria (MICs: 31.25–125 µg/ml). The pure compound (Ka) was combined with ZnO-NPs to improve effectiveness and reduce dose using checkerboard microdilution. Niteen treatments of Ka and ZnO-NPs combinations obtained by checkerboard matrix inhibited Klebsiella pneumonia. Eleven combinations had fractional inhibitory concentration index (FICi) between 1.03 and 2, meaning indifferent, another five combinations resulted from additive FICi (0.625–1) and only one combination with FICi of 0.5, indicating synergy. In the case of methicillin-resistant S. aureus (MRSA), Ka-ZnO-NPs combinations yielded 23 treatments with varying degrees of interaction. The results showed eleven treatments with indifferent interaction, eight additive interactions, and two synergies with FICi of 0.5 and 0.375. The combinations that exhibited synergy action were transformed into a nanoemulsion form to improve their solubility and bioavailability. The HR-TEM analysis of the nanoemulsion revealed spherical oil particles with a granulated core smaller than 200 nm and no signs of aggregation. Effective dispersion was confirmed by DLS analysis which indicated that Ka-ZnO-NPs nanoemulsion droplets have an average size of 53.1 nm and a polydispersity index (PI) of 0.523. The killing kinetic assay assessed the viability of methicillin-resistant Staphylococcus aureus (MRSA) and K. pneumonia post-treatment with Ka-ZnO-NPs combinations either in non-formulated or nanoemulsion form. Results showed Ka-ZnO-NPs combinations show concentration and time-dependent manner, with higher efficacy in nanoemulsion form. The findings indicated that Ka-ZnO-NPs without formulation at MIC values killed K. pneumonia after 24 h but not MRSA. Our nanoemulsion loaded with the previously mentioned combinations at MIC value showed bactericidal effect at MIC concentration of Ka-ZnO-NPs combination after 12 and 18 h of incubation against MRSA and K. pneumonia, respectively, compared to free combinations. At half MIC value, nanoemulsion increased the activity of the combinations to cause a bacteriostatic effect on MRSA and K. pneumonia after 24 h of incubation. The free combination showed a bacteriostatic impact for 6 h before the bacteria regrew to increase log10 colony forming unit (CFU)/ml over the initial level. Similarly, the cytotoxicity study revealed that the combination in nanoemulsion form decreased the cytotoxicity against kidney epithelial cells of the African green monkey (VERO) cell line. The IC50 for Ka-ZnO-NPs non-formulated treatment was 8.17/1.69 (µg/µg)/ml, but in nano-emulsion, it was 22.94 + 4.77 (µg/µg)/mL. In conclusion, efficient Ka-ZnO-NPs nanoemulsion may be a promising solution for the fighting of ESKAPE pathogenic bacteria according to antibacterial activity and low toxicity.

Intact NMR Approach Quickly Reveals Synchronized Microstructural Changes in Oil-in-Water Nanoemulsion Formulations

A soft-core oil-in-water (o/w) nanoemulsion (NE) is composed of nanometer (nm) sized oil droplets, stabilized by a surfactant layer and dispersed in a continuous bulky water phase. Characterization of the o/w NE molecule arrangements non-invasively, particularly the drug phase distribution (DPD) and its correlation to oil globule size (OGS), remains a challenge. Here we demonstrated the analytical methods of intact 19F Nuclear Magnetic Resonance (NMR) and 1H diffusion ordered spectroscopy (DOSY) NMR for their specificity in measuring DPD and OGS, respectively, on three NE formulations containing the active ingredient difluprednate (DFPN) at the same concentration. The results illustrated synchronized molecular rearrangement reflected in the DPD and OGS upon alterations in formulation. Addition of surfactant resulted in a higher DPD in the surfactant layer, and concomitantly smaller OGS. Mechanic perturbation converted most of the NE globules to the smaller thermodynamically stable microemulsion (ME) globules, changing both DPD and OGS to ME phase. These microstructure changes were not observed using 1D 1H NMR; and dynamic light scattering (DLS) was only sensitive to OGS of ME globule in mechanically perturbed formulation. Collectively, the study illustrated the specificity and essential role of intact NMR methods in measuring the critical microstructure attributes of soft-core NE systems quickly, accurately, and non-invasively. Therefore, the selected NMR approach can be a unique diagnostic tool of molecular microstructure or Q3 property in o/w NE formulation development, and quality assurance after manufacture process or excipient component changes.Graphical abstract

Ophthalmic Nanoemulsion Fingolimod Formulation for Topical Application.

Purpose: Fingolimod (FTY720; FT), a structural analog of sphingosine, has potential ocular applications. The goal of this study was to develop an FT-loaded nanoemulsion (NE; FT-NE) formulation for the efficient and prolonged delivery of FT to the posterior segment of the eye through the topical route. Methods: FT-NE formulations were prepared using homogenization followed by the probe sonication method. The lead FT-NE formulations (0.15% and 0.3% w/v loading), comprising soybean oil as oil and Tween® 80 and Poloxamer 188 as surfactants, were further evaluated for in vitro release, surface morphology, filtration sterilization, and stability at refrigerated temperature. Ocular bioavailability following topical application of FT-NE (0.3%) was examined in Sprague-Dawley rats. Results: The formulation, at both dose levels, showed desirable physicochemical characteristics, a nearly spherical shape with homogenous nanometric size distribution, and was stable for 180 days (last time point checked) at refrigerated temperature postfiltration through a polyethersulfone (0.22 µm) membrane. In vitro release studies showed prolonged release over 24 h, compared with the control FT solution (FT-S). In vivo studies revealed that effective concentrations of FT were achieved in the vitreous humor and retina following topical application of FT-NE. Conclusions: The results from these studies demonstrate that the FT-NE formulation can serve as a viable platform for the ocular delivery of FT through the topical route.

Formulation of nanoemulsion parijoto fruit extract (Medinilla Speciosa) with variation of tweens stabilizers.

Nanotechnology has substantial potential for development due to its ability to modify surface characteristics and particle size, facilitating enhanced absorption of functional food compounds and controlled release of active substances to mitigate adverse effects. Nanoemulsion, a stable colloidal system formed by blending oil, emulsifier, and water, was identified as nanotechnology with promising applications. However, investigations into the impact of surfactants on characteristic nanoemulsions need to be more varied. This research gap necessitated further exploration in the advancement of nanotechnology-based foods. The parijoto fruit (Medinilla speciosa), an indigenous plant species in Indonesia, has yet to undergo extensive scrutiny for its potential use as a functional and nutraceutical food. Anthocyanins, a principal compound in the parijoto fruit, had exhibited efficacy in reducing the risk of cardiovascular disease diabetes, demonstrating anti-inflammatory and antioxidant properties. This study aimed to investigate the characteristics of nanoemulsion formulations derived from parijoto fruit extract and to evaluate an optimum condition with various tween surfactants. The findings from this investigation could furnish valuable insights for the further advancement of anthocyanin nanoemulsions from parijoto fruit extract. The results comprised the characterization of nanoemulsion particle size, polydispersity index, ζ-potential, conductivity, pH, and viscosity. Through mathematical modeling and statistical methods, RSM optimizes nanoemulsion by examining the relationships and interactions between independent and response variables. Furthermore, the characterization of nanoemulsion encompassed ζ-potential, polydispersity, particle size, conductivity, pH, and viscosity. Elevated surfactant concentrations resulted in diminished particle sizes and more uniform size distribution, albeit reaching a plateau where surfactant aggregation and micelle formation ensued. Increased concentrations of surfactant type, concentration, and parijoto extract impacted the physical characteristics of nanoparticle size and polydispersity. The optimal process conditions for nanoemulsion consisting of the type of Tween used are Tween 80, Tween concentration of 12%, and parijoto fruit extract concentration of 7.5%, yielding a desirability value of 0.74, categorizing it as moderate.