High-speed Homogenization Research Articles

Protecting Hand-Knotted Carpets with Microbial Resistance

The carpet is a textile material known for its aesthetic appeal and is widely used for enhancing home and workplace décor. The hand-knotted carpets made from natural fibres like cotton tend to allow microbial growth when exposed to hot and humid conditions. This study involves the application of anti-microbial finishing chemicals developed from natural oils like Neem, Basil, and Aloe vera. Nano-emulsion of essential oil was prepared with polyoxyethylene sorbitan monooleate as a surfactant using a high-speed Homogenizer at 15K rpm for 30 minutes. These essential oils can repel insects like house flies and prevent the growth of microbes which cause human health-related issues like skin itching, irritation, sneezing and allergy. Varying concentrations of emulsion oil 1% and 5% were used to prepare nano-emulsion formulation, and the particle size and thermal stability were evaluated. FTIR analysis of the emulsion was done to check the presence of functional groups. The spraying technique was used for the application of neem oil emulsion on the carpet and dried. The finished carpet was tested for the anti-microbial performance effect and found to provide adequate microbial protection.

Development and Optimization of Enzalutamide Nanosuspension by Design of Experiments for Dissolution Enhancement

Enzalutamide is an anticancer molecule used for prostate cancer. The goal of the study was to develop a nanosuspension of enzalutamide in order to improve its solubility and dissolution properties. High-speed homogenization method was employed to formulate the nanosuspension. Preliminary studies suggested the amount of stabilizer, homogenization time and homogenization speed as critical variables to be taken for the optimization process. Box-Behnken design was employed for the optimization of process and formulation variables. Nanosuspension was evaluated for particle size, PDI, zeta potential, and in-vitro drug release at 10 minutes (D10) studies. Regression analysis suggested the influence of independent variables on the responses. The optimized batch obtained from the overlay plot exhibited 198.36 nm of particle size, (-33.27 mV of zeta potential and 80.47% of D10 values). The characterization studies i.e., DSC, and XRD illustrated retention in crystallinity of the drug. The drug and formulation were found to be stable over a 6-month period in accelerated stability testing. Using high speed homogenization method, the particle size of the formulation was reduced to nano-size, which was further responsible for the improvement in dissolution and bioavailability of the drug.

Cellulose Nanocrystal-Based Emulsion of Thyme Essential Oil: Preparation and Characterisation as Sustainable Crop Protection Tool

Essential oil-based pesticides, which contain antimicrobial and antioxidant molecules, have potential for use in sustainable agriculture. However, these compounds have limitations such as volatility, poor water solubility, and phytotoxicity. Nanoencapsulation, through processes like micro- and nanoemulsions, can enhance the stability and bioactivity of essential oils. In this study, thyme essential oil from supercritical carbon dioxide extraction was selected as a sustainable antimicrobial tool and nanoencapsulated in an oil-in-water emulsion system. The investigated protocol provided high-speed homogenisation in the presence of cellulose nanocrystals as stabilisers and calcium chloride as an ionic crosslinking agent. Thyme essential oil was characterised via GC-MS and UV-vis analysis, indicating rich content in phenols. The cellulose nanocrystal/essential oil ratio and calcium chloride concentration were varied to tune the nanoemulsions’ physical–chemical stability, which was investigated via UV-vis, direct observation, dynamic light scattering, and Turbiscan analysis. Transmission electron microscopy confirmed the nanosized droplet formation. The nanoemulsion resulting from the addition of crosslinked nanocrystals was very stable over time at room temperature. It was evaluated for the first time on Pseudomonas savastanoi pv. savastanoi, the causal agent of olive knot disease. In vitro tests showed a synergistic effect of the formulation components, and in vivo tests on olive seedlings demonstrated reduced bacterial colonies without any phytotoxic effect. These findings suggest that crosslinked cellulose nanocrystal emulsions can enhance the stability and bioactivity of thyme essential oil, providing a new tool for crop protection.

Impact of High-Speed Homogenisation Followed by pH Treatment of Arthrospira platensis on Protein Accessibility and In Vitro Protein Digestibility

Impact of High-Speed Homogenisation Followed by pH Treatment of Arthrospira platensis on Protein Accessibility and In Vitro Protein Digestibility

Elastic 3D-Printed Nanofibers Composite Scaffold for Bone Tissue Engineering.

Loading nanoparticles into hydrogels has been a conventional approach to augment the printability of ink and the physicochemical characteristics of scaffolds in three-dimensional (3D) printing. However, the efficacy of this enhancement has often proven to be limited. We amalgamate electrospun nanofibers with 3D printing techniques to fabricate a composite scaffold reminiscent of a "reinforced concrete" structure, aimed at addressing bone defects. These supple silica nanofibers are synthesized through a dual-step process involving high-speed homogenization and low-temperature ball milling technology. The nanofibers are homogeneously blended with sodium alginate to create the printing ink. The resultant ink was extruded seamlessly, displaying commendable molding properties, thereby yielding scaffolds with favorable macroscopic morphology. In contrast to nanoparticle-reinforced scaffolds, composite scaffolds containing nanofibers exhibit superior mechanical attributes and bioactivity. These nanofiber composite scaffolds demonstrate enhanced osteoinductive properties in both in vitro and in vivo evaluations. To conclude, this research introduces a novel 3D printing approach where the fabricated nanofiber-infused 3D-printed scaffolds hold the potential to revolutionize the realm of 3D printing in the domain of bone tissue engineering.

Emamectin-sodium alginate nano-formulation based on charge attraction with highly improved systemic translocation and photolysis resistance

Nano pesticides offer an effective means of improving the bioavailability of pesticide due to their excellent solubility and wettability, superior foliar adhesion, and permeability to target insects. By using high-speed homogenization and ultrasonic dispersion technology, an emamectin-sodium alginate nano-formulation (EB@SA) with a particle size ranging from 30 to 50 nm was successfully fabricated using electrostatic self-assembly. The microscopic morphology and structure of EB@SA were further analyzed through transmission electron microscopy, dynamic light scattering, infrared spectroscopy, and 1H NMR. The photolysis resistance behavior of EB@SA demonstrated an improved anti-photolysis ability more than double that of conventional formulations while also exhibiting good sustained-release properties. Not only does EB@SA maintain the inherent insecticidal toxicity of emamectin benzoate (EB), but it also significantly prolongs its insecticidal duration. At a concentration of 20 mg/L, the lethality rate against Armyworms remains above 70 % over a period of 16 days compared to <50 % for general emamectin emulsifiable concentrate. Furthermore, EB@SA greatly enhances the systemic translocation of EB in corn plants by exhibiting favorable bidirectional systemic translocation characteristics. This research presents an efficient and environmentally friendly pesticide nano-formulation that can be effectively utilized for field pest control.

Effect of violent mixing on sonochemical oxidation activity under various geometric conditions in 28-kHz sonoreactor

The effects of violent mixing and reactor geometric conditions were investigated using the overhead stirrer and high-speed homogenizer in 28-kHz sonoreactors. The sonochemical oxidation activity was quantified using the KI dosimetry method, and the sonochemical active zone was visually observed using the luminol method. Higher mixing rates resulted in a significant enhancement of the sonochemical oxidation activity, primarily due to a significant change in the sonochemical active zone. When using the overhead stirrer (0–2,000 rpm), the highest activity for 2λ and 3λ occurred at 500 rpm, whereas the highest activity for 4λ was obtained at 250 rpm. For the high-speed homogenizer (0–12,000 rpm), the highest activity was consistently obtained at 3,500 rpm across all liquid height conditions. The impact of mixing position (Top, Mid, and Bot positions) on sonochemical activity was analyzed. The results revealed that the lowest activity was obtained for the bottom position, likely attributed to significant ultrasound attenuation. The reactor size effect was investigated using the high-speed homogenizer in five cylindrical sonoreactors with different diameters (12–27 cm). It was found that very low activity could be observed due to unexpected geometric conditions, and the application of mixing (3,500 rpm in this study) could result in high sonochemical activity regardless of geometric conditions.

Adipokine-Enriched Adipose Extract Restores Skin Barrier and Ameliorates Inflammatory Dysregulation in Atopic Dermatitis Mice.

Atopic dermatitis (AD) is a chronic dermatosis with high incidence worldwide characterized by skin barrier abnormalities and immune dysregulation. Conventional therapies are usually limited by side effects and high cost. Given the antiinflammatory and repairing properties, adipokines are increasingly considered as promising therapeutic agents for dermatoses. Adipose collagen fragments (ACF), a novel adipokine-enriched product, may alleviate AD through modulating immune microenvironment and restoring skin barrier. ACF was extracted from adipose tissue by means of high-speed homogenization (10,000 rpm/min for 1 minute) and centrifugation (3000 g for 3 minutes). Ovalbumin-induced AD female BALB/c mice (6-week-old) were intradermally injected with 0.2 mL of ACF or phosphate-buffered saline (negative control), with normal mice being set as normal control ( n = 6). Dermatitis severity, inflammatory metrics (epidermal thickness, infiltrated mast cells, T helper cell [Th]-type cytokine expression), and skin barrier-related metrics (transepidermal water loss, skin barrier-related proteins expression) were evaluated after the AD induction period (day 50). ACF-derived bioactive components were also evaluated using proteomic analysis. ACF-derived adipokines contained antiinflammatory, skin barrier- and lipid biosynthesis-related components. ACF treatment decreased dermatitis severity (6.2 ± 1.8 [ P < 0.0001]), epidermal thickness (25.7 ± 12.8 μm [ P = 0.0045]), infiltrated mast cells (31.3 ± 12.4 cells/field [ P = 0.0475]), and expression of Th-type cytokines (interferon-γ, tumor necrosis factor-α, interleukin [IL]-4, IL-4R, IL-13, and IL-17A [ P < 0.05]) in AD skins. Transepidermal water loss (29.8 ± 13.8 g/m 2 per hour [ P = 0.0306]) and skin barrier-related protein expression (filaggrin, 14,258 ± 4375 [ P = 0.0162]; loricrin, 6037 ± 1728 [ P = 0.0010]; claudin-1, 20,043 ± 6406 [ P = 0.0420]; and zonula occludens-1, 4494 ± 1114 [ P = 0.0134]) were also improved. ACF improved AD in a murine model by ameliorating inflammatory dysregulation and skin barrier defects. Further validation is needed in more advanced animal models. ACF is an injectable, adipose-derived collagen scaffold prepared from autologous harvested fat using fast and simple mechanical methods. ACF may reduce the limitations associated with health care regulatory issues and serve as a promising autologous therapeutic agent for skin disorders in clinics.

Chia Oil Microencapsulation Using Tannic Acid and Soy Protein Isolate as Wall Materials.

The use of proteins to produce oil-containing microcapsules has been previously analyzed; however, their chemical modification, in order to improve their performance as wall materials, is a strategy that has not been widely developed yet. This study aimed to analyze the chemical modification of the proteins through cross-linking reactions with tannic acid and to evaluate their performance as wall materials to the microencapsulation of oils rich in polyunsaturated fatty acids. The cross-linking reaction of isolated soy protein and tannic acid was carried out at pH 10-11 and 60 °C. Subsequently, emulsions were made with a high-speed homogenizer and microcapsules were obtained by spray drying. Microcapsules were characterized by particle size, morphology (SEM), total pore area and % porosity (mercury intrusion methodology), superficial properties (contact angle), and size distribution of oil droplets (by laser diffraction). Additionally, encapsulation efficiency was determined as a function of total and surface oil. Oil chemical stability and quality were studied by Rancimat, hydroperoxide values, and fatty acid profiles. In addition, a storage test was performed for 180 days, and released oil and polyphenols were determined by in vitro gastric digestion. Moreover, the fatty acid composition of the oil and the total polyphenol content and antioxidant capacity of polyphenols were analyzed. The results showed that spray-dried microcapsules had an encapsulation efficiency between 54 and 78%. The oxidative stability exhibited a positive correlation between the amount of polyphenols used and the induction time, with a maximum of 27 h. The storage assay showed that the peroxide value was lower for those cross-linked microcapsules concerning control after 180 days. After the storage time, the omega-3 content was reduced by 49% for soy protein samples, while cross-linked microcapsules maintained the initial concentration. The in-vitro digestion assay showed a decrease in the amount of oil released from the cross-linked microcapsules and an increase in the amount of polyphenols and a higher antioxidant capacity for all samples (for example, 238.10 mgGAE/g and 554.22 mg TE/g for undigested microcapsules with TA 40% versus 322.09 mgGAE/g and 663.61 mg TE/g for digested samples). The microcapsules showed a high degree of protection of the encapsulated oil, providing a high content of polyunsaturated fatty acids (PUFAS) and polyphenols even in prolonged storage times.

Green Synthesis of Low-Glycemic Amylose-Lipid Nanocomposites by High-Speed Homogenization and Formulation into Hydrogel.

In this research, we focused on the production of amylose-lipid nanocomposite material (ALN) through a green synthesis technique utilizing high-speed homogenization. Our aim was to investigate this novel material's distinctive physicochemical features and its potential applications as a low-glycemic gelling and functional food ingredient. The study begins with the formulation of the amylose-lipid nanomaterial from starch and fatty acid complexes, including stearic, palmitic, and lauric acids. Structural analysis reveals the presence of ester carbonyl functionalities, solid matrix structures, partial crystallinities, and remarkable thermal stability within the ALN. Notably, the ALN exhibits a significantly low glycemic index (GI, 40%) and elevated resistance starch (RS) values. The research extends to the formulation of ALN into nanocomposite hydrogels, enabling the evaluation of its anthocyanin absorption capacity. This analysis provides valuable insights into the rheological properties and viscoelastic behavior of the resulting hydrogels. Furthermore, the study investigates anthocyanin encapsulation and retention by ALN-based hydrogels, with a particular focus on the influence of pH and physical cross-link networks on the uptake capacity presenting stearic-acid (SA) hydrogel with the best absorption capacity. In conclusion, the green-synthesized (ALN) shows remarkable functional and structural properties. The produced ALN-based hydrogels are promising materials for a variety of applications, such as medicine administration, food packaging, and other industrial purposes.

Fine-tuning formulation and biological interaction of doxorubicin-loaded polymeric nanoparticles via electrolyte concentration modulation

Doxorubicin (DOX) is amongst the most widely used chemotherapeutic drugs against various cancers. However, its controlled biodistribution to the tumor site at the required pharmacokinetics profile remains challenging. In this work, a vast study of formulation parameters has been performed to control doxorubicin loading into polymeric nanoparticles in physiological conditions. Water-in-oil-in-water (W1/O/W2) template emulsions have been formulated, including doxorubicin in the internal water phase. Specifically, the system [Doxorubicin in aqueous solution (W1) / Pluronic F-127 (S) and PLGA in ethyl acetate (O)], [Polysorbate 80 (S) in water solution], using phosphate buffer on both aqueous phases at different electrolyte concentrations has been established. The first W1/O nano-emulsion was formed by high-speed homogenization, and the second O/W2 nano-emulsion was formed by the phase inversion composition method, a low-energy emulsification appropriated for pharmaceutical compounds. Although many previous studies had already used double emulsions for DOX encapsulation in nanofluids, here, for the first time, the salt concentration of the water phases has been varied to control the resulting doxorubicin and nanoparticle properties. The addition of high concentrations of electrolytes in both aqueous phases decreases the solubility and dissolution rates of the drug, leading to a high degree of DOX dimerization. This phenomenon significantly impacts the internalization and subcellular localization. Facilitating drug nuclear accumulation is of vital interest, and this is primarily achieved when doxorubicin is incorporated into nanoparticles formulated with PBS at low electrolyte concentrations (2.5 mM), which exhibit exceptional internalization properties. Thus, it has been demonstrated that doxorubicin loading in polymeric nanoparticles can be tuned by only varying salt concentrations of template W1/O/W2 emulsions. Consequently, our results show the adequacy of the double nano-emulsion templating for the generation of non-toxic DOX-loaded polymeric nanoparticles with different physicochemical properties and interaction with the cell surface that would allow their use for various anticancer therapies.

Shear emulsification condition strategy impact high internal phase Pickering emulsions stabilized by coconut globulin-tannic acid: Structure of protein at the oil-water interface

High internal phase Pickering emulsions (HIPPEs) have gained increasing attention due to their exceptional stability. However, the stability of HIPPEs relies on specific preparation conditions. In this research, the effects of high-speed homogenization conditions on HIPPEs stabilized by coconut globulin-tannic acid (CG-TA) colloidal particles were investigated. CG-TA colloidal particles exhibited optimal wetting properties (83.5°) and created a dense interface layer (44 nm). Insufficient shearing could not effectively homogenize oil and water phases, resulting in larger droplet sizes (33 μm). Appropriately applied shearing, especially at 10000 r/min for 1 min, facilitated the reduction of α-helix content to 38% and unfolding of CG molecule, resulting in a decrease in droplet size. Meanwhile, droplet size change of HIPPEs was minimum in 28 days and profiles difference was the minimal after centrifugation. Nevertheless, excessive shearing destroyed the hydrogen bond (O–H and N–H bending vibrations shifted to 3417 cm−1) between particles, causing an increase in particle size to 369 nm and a decline in adsorbed protein percentage to 64%. Therefore, these results proved that the shear rate and time were important to the stability of protein-based HIPPEs during homogenization, thus providing theoretical basis for the application of HIPPEs, such as solid fat substitutes.

Nanostructured Lipid Carrier of Cinacalcet HCl: Formulation, BBD Enabled Optimization, Pharmacokinetic and In-Vitro Cytotoxicity Study

Background: Cinacalcet hydrochloride (CINH) is a BCS class IV drug. It is mainly used for the treatment of chronic renal disease and parathyroid cancer. It exhibits poor oral bioavailabil-ity of less than 25%. Objectives: The main objective is to improve the bioavailability of CINH by formulating the nanostructure lipid carrier (NLC). Methods: In this research, glyceryl monostearate (GMS), labrasol, and tween 20 were the main ex-cipients selected for the formulation of NLC. Hot high-speed homogenization and ultra-sonication method was used for the NLC formulation of CINH. The characterization of the NLCs was done as per standard procedures. Optimization of the formulated NLC was carried out by applying Box-Behnken Design (BBD) with the help of the Design Expert software. The pharmacokinetic study was conducted to determine the improvement in the bioavailability of the CINH. The cytotoxicity study was performed by using the MTT assay method to know the cell viability. Results: The optimized NLC formulation exhibited high drug content with a particle size of less than 200nm. A pharmacokinetic study showed 4 fold increase in oral bioavailability for the opti-mized NLC in comparison to the aqueous suspension of CINH. Minimum viability was determined as 94%, which indicates the safety of the incubated formulations. Conclusion: NLC formulation has the potential to improve oral bioavailability with high drug load-ing and cell viability for CINH.

Easy-process nanoemulsions: Obtaining thymol nanodroplets with high shear speed systems

Several mechanisms to form nanoemulsions are reported in the literature. However, there are still gaps in the relationship between the process variables with physicochemical and bioactive properties. This study applied a central composite rotatable design (CCRD) to describe the processing variables of a high-speed homogenizer (rotor/stator) on the physicochemical properties of a thymol nanoemulsion. Droplet sizes of 68.47 nm, 0.112 PDI, and −11.00 mV zeta potential could be obtained with improved conditions of 1.0% thymol, 4.63% Tween 80, 6600 rpm, and 3.13 min rotation. The improved nanoemulsions stored at 4 °C under different conditions (NaCl 0.5%, NaCl 1.0%, pH 4.5, and pH 7.5) showed droplet size stability over 45 days (<200 nm). In addition, the improved thymol nanoemulsions showed better antibacterial activity than non-nanoemulsified thymol for all clinically significant pathogens studied (Staphylococcus aureus, Listeria monocytogenes, Pseudomonas aeruginosa, Escherichia coli, and Salmonella Enteritidis). The properties of nanodroplets observed in this study highlight the use of high-speed homogenizers as a viable alternative in the development of nanoemulsions.

Characterization and encapsulation efficiency of zein nanoparticles loaded with chestnut fruit shell, cedar and sweetgum bark extracts

Zein nanoparticles (ZNPs) loaded with bioactive extracts of chestnut (Castanea sativa Mill.) shell, cedar (Cedrus libani) and sweetgum (Liquidambar orientalis) bark wastes were produced using different methods. Nanoprecipitation, high-speed homogenization and ultrasonic homogenization allowed the fabrication of ZNPs with particle sizes smaller than 202.40 nm, 430.25 nm and 325.50 nm, respectively. The smallest nanoparticle size was achieved at 132.81 nm for sweetgum bark extract-loaded ZNPs obtained by the nanoprecipitation method. Encapsulation efficiency (EE) was between 34.03 and 96.83% for all zein nanoparticles fabricated under different mixtures and process conditions. Zein concentration and extract ratio played an essential role in the EE of nanoparticles. The best conditions were determined to obtain the desired properties of ZNPs based on particle size, polydispersity index and EE by using a central composite rotatable design. The nanoprecipitation method was more appropriate for producing chestnut and cedar shell/bark extract-loaded nanoparticles. In contrast, the high-speed homogenization method was suitable for producing sweetgum bark extract-loaded nanoparticles. As a result of the encapsulation of various shell/bark extracts within zein nanoparticles, value-added products were generated from wastes having bioactive compounds. The developed zein nanoparticles for each extract type would offer eco-friendly, simple and safe food processing and packaging systems.

Development and characterization of a continuous ultrasound emulsification and nano-emulsion polymerization process

Typically, nano-emulsions are prepared in batch high-speed homogenization or ultrasound processes and polymerized afterwards in batch reactors. However, fully continuous processes have the potential to decrease production costs and energy consumption compared to batch processes. This research thus focuses on implementing ultrasound into a fully continuous emulsification and nano-emulsion polymerization process to obtain latex nano-particles from butylmethacrylate and ethylene glycol dimethacrylate. The goal of this research is to produce nano-emulsions and nano-particles with the smallest obtainable size and polydispersity in a fully continuous two-stage process. In the first stage or emulsification stage, parameters like flowrate, residence time and acoustic power are varied to influence and determine the optimal energy density. In second stage or reaction stage, residence time and reaction temperature are studied to obtain particles of monodisperse sizes. Samples produced in both stages are analysed with Dynamic Light Scattering to measure the average size and polydispersity (PdI) of the emulsion droplets and particles. Emulsification results indicate that droplet size and PdI decreases at increasing energy densities (J/ml) until 60 J/ml after which a constant droplet size of 150 nm and PdI of 0.230 are reached. Furthermore, a particle size of 50 nm and PdI of 0.080 were achieved in both batch and continuous polymerization reactors after 5 min at 85 °C. By implementing the most optimal process parameters, small emulsion droplets and particles were obtained more energy efficient in a shorter emulsification and reaction time compared to literature ultrasound assisted nano-emulsion polymerization processes.

Development of lipid nanoparticles with nystatin for an antifungal action

Fungal diseases currently affect about a quarter of the population worldwide. Fungi infections caused by Candida albicans have been described as a significant concern to public health. The spectrum of clinical diseases caused by this fungi species range between vulvovaginal candidiasis, oral candidiasis, candidemia and mucositis. The emergence of resistance mechanisms towards antifungal therapy greatly hampers successful management of illness and patient outcome. Nystatin, an antifungal drug, is categorized as a class IV of Biopharmaceutical Classification System, presenting low aqueous solubility and low intestinal permeability. Nowadays, the emerging platform of nanotechnology and lipid nanoparticles, notably solid lipid nanoparticles (SLN), has been subject to growing attention over recent past, owing to the promising properties of vectorization among a substantial variety of pharmaceutical drugs. Due to its hydrophobic proprieties, nystatin was encapsulated in SLN. Thus aiming to understand the relationship between the use of nanosystems and the improvement of the therapeutic effect. The aim of this work was to formulate SLN with nystatin by different methods (high speed homogenization and ultrasonication) with optimization of several parameters and formulation of 2 gels (one of them containing nanoparticles).&#x0D; Initially, 3 lipids were used: Compritol® 888 ATO, cetyl palmitate and Precirol® ATO 5 and, after the study of several parameters (size, encapsulation efficiency (EE) and polymorphic behaviour of the lipids), Precirol® ATO 5 was chosen as the lipid with the most satisfactory results. The results of the present work showed that the assay method of nystatin was linear, specific and presented repeatability. The average diameter of empty nanoparticles (NPs) and with drug (Precirol-NYS NPs) was, respectively, 306 nm and 260 nm and an EE of 67.8%. Regarding stability, SLN with drug proved to be more stable than SLN without drug. The polymer used for formulation of gels was the polymer commonly known by the trade name Carbopol® 940. The yield of 0.5% Carbopol® gel preparations and 0.5% Carbopol® gel + 10% Precirol-NYS NPs were 87.2% and 91.39%, respectively.

Encapsulation of olive leaf extract using double emulsion method

Olive leaves are obtained as a waste product of the olive industry. Biophenols, abundantly found in olive leaves, are susceptible to heat, light, and oxidizing agents, which necessitates encapsulation to increase their bioavailability. In this study, the double emulsion method was preferred due to its protective effect on the active substance and the control over its release. The effects of different pea flour concentrations (15%, 20%, and 25%) used in outer aqueous phase of double emulsion and homogenization methods [high-speed homogenization (HSH) and ultrasonication (US)] on emulsion properties were investigated. The particle size, rheology, encapsulation efficiency, stability, optical images, and release behavior of the emulsions were determined. As hypothesized, flours acted as emulsifiers in the outer aqueous phase to increase the stability of emulsions. It was observed that the stability of emulsions was correlated with the viscosity and particle size. Increasing pea flour concentration from 15% to 25% resulted in a 25% and 30% increase in the stability of double emulsions prepared with HSH and US, respectively. The higher stability of emulsions prepared with 25% was due to their higher viscosity and smaller particle size. Samples were found to have shear-thinning behavior. Moreover, emulsions stored at 20 °C showed faster degradation compared to 4 °C. US treatment did not decrease the average particle size of emulsions. Average encapsulation efficiency for double emulsions prepared with HSH and US was 88.3% and 85.9%, respectively. As a result, pea flours could be used to encapsulate olive leaf extract successfully with high encapsulation efficiencies by using the double emulsion method.

Topical caffeine-loaded nanostructured lipid carriers for enhanced treatment of cellulite: A 32 full factorial design optimization and in vivo evaluation in rats

The goal of this study was the development and evaluation of semisolid caffeine (CAF) loaded nanostructured lipid carriers (NLCs) for topical treatment of cellulite. CAF-loaded NLC formulations were prepared via high-speed homogenization followed by ultrasonication. A 32 full factorial design was employed for formulation optimization. The total lipid content (%) and the liquid lipid content per total lipids (%) were chosen as factors, whereas particle size (PS), polydispersity index (PDI), zeta potential (|ZP|) and viscosity (VIS) were selected as responses. The design suggested CAF-NLC3 as the optimum formulation consisting of a total lipid content of 15% w/w (palmitic acid and soft paraffin/isopropyl myristate, 7:3 w/w) and a surfactant content of 10% w/w (Tween 80/lecithin, 8:1.2 w/w). CAF-NLC3 revealed PS, PDI, ZP, VIS and CAF content values of 318.8 nm, 0.253, −41.1 mV, 18.0 Pa.s and 97.57%, respectively. It showed a pseudoplastic rheological behavior, acceptable pH value (5.25), good spreadability (1.12 mm2/g) and spherical shape employing transmission electron microscopy. Differential scanning calorimetry and X-ray diffraction demonstrated the amorphization of CAF in CAF-NLC3. CAF-NLC3 remained stable for 3 months at room and refrigeration conditions. A single topical application of CAF-NLC3 on shaved abdominal skins of Wistar rats revealed enhanced skin retention of CAF by 2-fold and 1.4-fold after 4 h when compared with plain CAF gel (CAF-P) and marketed CAF gel (CAF-M), respectively. Furthermore, CAF-NLC3 exhibited a superior anti-cellulite activity in comparison with CAF-P and CAF-M through elevating extracellular matrix components (collagen 1, elastin and hyaluronic acid) and stimulating the brown adipose tissue thermogenesis via up-regulating UCP1 and PPAR-γ expression. In addition, CAF-NLC3 prominently increased lipolysis through HSL activity and decreased pro-inflammatory cytokines such as ICAM-1 and VCAM-1 after 30 days of treatment on a high fat diet-induced cellulite rat model. These findings were further confirmed by histopathological examination supported by morphometric analysis. Therefore, incorporation of CAF in a semisolid NLC formulation would be a promising cosmetic approach for the topical treatment of cellulite.

A quick, easy and efficient protocol for extracting high-quality RNA from Mycobacterium tuberculosis using a spin column commercial kit

RNA extraction from Mycobacterium tuberculosis has been a historically challenging task for researchers due to the thick lipids associated with the cell wall of this “notorious” pathogen that is responsible for Tuberculosis (TB) outbreaks. Several studies have successfully extracted RNA from M. tuberculosis using a Trizol reagent combined with organic solvents. Recently, our laboratory has successfully extracted high quality total RNA using a commercial kit from clinical strains belonging to F15/LAM4/KZN, Beijing and F11 strain families and H37Rv laboratory strain by exploiting high speed homogenizer for cell lysis and spin columns for RNA purification. The quality and integrity of the extracted RNA was analyzed and confirmed through the Nanodrop, Bioanalyzer and RNA 3-(N-morpholino) propanesulfonic acid (MOPS) gel electrophoresis. Furthermore, to confirm the integrity of small RNA (sRNA) molecules due to their vulnerability to degradation, the RNA samples were converted to cDNA and sRNAs were amplified and confirmed through PCR. This detailed RNA extraction protocol proposes to carve a new path into TB transcriptome research without the use of organic solvent for downstream purification steps while yielding high quality RNA that can be used to understand M. tuberculosis transcriptome regulation.